Factors affecting progression of knee cartilage defects in normal subjects over 2 years.
Cartilage defects are present in subjects with knee osteoarthritis (OA). Although they are often present in healthy subjects, there is little data on the natural history of cartilage defects. The aim of this study was to examine the change in cartilage defects over 2 yr and to identify factors associated with this change. One hundred and twenty-four healthy subjects underwent magnetic resonance imaging of their dominant knee at baseline and follow-up. Cartilage defects were scored (0-4) at five sites. Bone size was determined at medial and lateral tibial plateau and patella. Height, weight, body mass index and physical activity were measured by standard protocols. Eighty-six subjects completed the study. The mean cartilage defect score of each tibiofemoral compartment increased over time. However, medial and lateral tibiofemoral defect score decreased in 5% of the subjects. Cartilage defects were more likely to progress in males than females in each individual compartment (P<0.001 for medial tibiofemoral, P=0.005 for lateral tibiofemoral and P=0.01 for patellar cartilage). Baseline cartilage defect score was negatively associated with the progression of cartilage defects in each compartment (all P<0.001). Although knee cartilage defects progressed over time in the majority of normal subjects, those of the highest severity tended to regress. Male gender and baseline cartilage defect score were the main factors associated with the progression of cartilage defects. Larger studies will be required to identify factors associated with the progression and regression of lesions.


All-solid-state flexible supercapacitor based on nanotube-reinforced polypyrrole hollowed structures.
Supercapacitors are strong future candidates for energy storage devices owing to their high power density, fast charge-discharge rate, and long cycle stability. Here, a flexible supercapacitor with a large specific capacitance of 443 F g-1 at a scan rate of 2 mV s-1 is demonstrated using nanotube-reinforced polypyrrole nanowires with hollowed cavities grown vertically on a nanotube/graphene based film. Using these electrodes, we obtain improved capacitance, rate capability, and cycle stability for over 3000 cycles. The assembled all-solid-state supercapacitor exhibits excellent mechanical flexibility, with the capacity to endure a 180° bending angle along with a maximum specific and volumetric energy density of 7 W h kg-1 (8.2 mW h cm-3) at a power density of 75 W kg-1 (0.087 W cm-3), and it showed an energy density of 4.13 W h kg-1 (4.82 mW h cm-3) even at a high power density of 3.8 kW kg-1 (4.4 W cm-3). Also, it demonstrates a high cycling stability of 94.3% after 10 000 charge/discharge cycles at a current density of 10 A g-1. Finally, a foldable all-solid-state supercapacitor is demonstrated, which confirms the applicability of the reported supercapacitor for use in energy storage devices for future portable, foldable, or wearable electronics.


The SbMT-2 gene from a halophyte confers abiotic stress tolerance and modulates ROS scavenging in transgenic tobacco.
Heavy metals are common pollutants of the coastal saline area and Salicornia brachiata an extreme halophyte is frequently exposed to various abiotic stresses including heavy metals. The SbMT-2 gene was cloned and transformed to tobacco for the functional validation. Transgenic tobacco lines (L2, L4, L6 and L13) showed significantly enhanced salt (NaCl), osmotic (PEG) and metals (Zn++, Cu++ and Cd++) tolerance compared to WT plants. Transgenic lines did not show any morphological variation and had enhanced growth parameters viz. shoot length, root length, fresh weight and dry weight. High seed germination percentage, chlorophyll content, relative water content, electrolytic leakage and membrane stability index confirmed that transgenic lines performed better under salt (NaCl), osmotic (PEG) and metals (Zn++, Cu++ and Cd++) stress conditions compared to WT plants. Proline, H2O2 and lipid peroxidation (MDA) analyses suggested the role of SbMT-2 in cellular homeostasis and H2O2 detoxification. Furthermore in vivo localization of H2O2 and O2-; and elevated expression of key antioxidant enzyme encoding genes, SOD, POD and APX evident the possible role of SbMT-2 in ROS scavenging/detoxification mechanism. Transgenic lines showed accumulation of Cu++ and Cd++ in root while Zn++ in stem under stress condition. Under control (unstressed) condition, Zn++ was accumulated more in root but accumulation of Zn++ in stem under stress condition suggested that SbMT-2 may involve in the selective translocation of Zn++ from root to stem. This observation was further supported by the up-regulation of zinc transporter encoding genes NtZIP1 and NtHMA-A under metal ion stress condition. The study suggested that SbMT-2 modulates ROS scavenging and is a potential candidate to be used for phytoremediation and imparting stress tolerance.


PsbS protein modulates non-photochemical chlorophyll fluorescence quenching in membranes depleted of photosystems.
Plants with varying levels of PsbS protein were grown on lincomycin. Enhanced levels of non-photochemical fluorescence quenching (NPQ) in over-expressers of the protein have been observed. This was accompanied by increased amplitude of the irreversible NPQ component, qI, previously considered to reflect mainly photoinhibition of PSII reaction centres (RCII). However, since RCIIs were largely absent the observed qI is likely to originate from the LHCII antenna. In chloroplasts of over-expressers of PsbS grown on lincomycin an abnormally large NPQ (∼7) was characterised by a 0.34 ns average chlorophyll fluorescence lifetime. Yet the lifetime in the Fm state was similar to that of wild-type plants. 77K fluorescence emission spectra revealed a specific 700 nm peak typical of LHCII aggregates as well as quenching of the PSI fluorescence at 730 nm. The aggregated state manifested itself as a clear change in the distance between LHCII complexes detected by freeze-fracture electron microscopy. Grana thylakoids in the quenched state revealed 3 times more aggregated LHCII particles compared to the dark-adapted state. Overall, the results directly demonstrate the importance of LHCII aggregation in the NPQ mechanism and show that the PSII supercomplex structure plays no role in formation of the observed quenching.


Bacteremia in hospitalized children.
The results of 6,951 blood cultures taken from hospitalized children are reviewed. One or more organisms grew from 6% (399) of the cultures, of which 189 (two thirds) were considered to represent confirmed bacteremia. The most common organisms associated with bacteremia were Haemophilus influenzae, Streptococcus pneumoniae, enterobacteriaceae, and Staphylococcus aureus. Patients with deficient host defenses (newborns, oncology patients) with bacteremia had a higher mortality than normal children. Laboratory techniques allowing more rapid detection of positive blood cultures resulted in major alterations in therapy in almost one half of all bacteremic patients.


Low-dose preoperative aspirin therapy, postoperative blood loss, and transfusion requirements.
The effects of three low-dose regimens of preoperative aspirin therapy on postoperative blood loss, transfusion requirements, and length of hospital stay were recorded in a prospective cohort study of 202 patients undergoing elective coronary artery bypass grafting. One hundred one patients had been prescribed daily aspirin by the referring cardiologist (44 at 75 mg, 28 at 150 mg, and 29 at 300 mg); the remaining 101 patients who had not been prescribed aspirin acted as a control group. A median postoperative blood loss of 870 mL in the control group was increased by 280 mL in the 75-mg aspirin group (p less than 0.001), by 490 mL in the 150-mg aspirin group (p less than 0.001), and by 230 mL in the 300-mg aspirin group (p = 0.03). The median requirement for blood transfusion of 2 U red blood cell concentrates in the control group was increased by 2 U in the 75-mg aspirin group (p less than 0.001), 2 U in the 150-mg aspirin group (p less than 0.001), and 1 U in the 300-mg aspirin group (p = 0.05). Hemostatic "packs" (fresh frozen plasma, platelets, and cryoprecipitate) were required in 20 patients in the aspirin groups as compared with 5 in the control group (p less than 0.01 by chi 2 test). The mean postoperative hospital stay was 8 days for all groups. Regular daily low-dose aspirin therapy produces significant increases in postoperative blood loss, resulting in a substantial increase in blood transfusion and hemostatic pack requirements, but does not prolong postoperative hospital stay.


What do patients with glaucoma think about personal health records?
Putting patients in control of their records is one way of promoting patient centred care and patients with chronic health problems may benefit most from personal health records (PHRs). Glaucoma management is often complicated by incomplete understanding and poor adherence to treatment, two areas which a PHR may help to address. This study aimed to discover what patients with glaucoma think about PHRs and what type of information a glaucoma PHR should contain. A consultation exercise using a focus group approach involving 71 participants was undertaken to discuss if a PHR would be useful and what it would be like. Narrative data were collected through written notes and an online forum in addition to transcripts of the focus group feedback session and individual interviews. Recordings were transcribed and analysed with simple thematic analysis facilitated by NVivo software (www.qsrinternational.com). The consultation exercise indicated enthusiasm for PHRs. Views varied on the best format, some participants strongly favoured electronic records and others preferred a low-tech, paper based format. A comprehensive dataset of 24 items was developed which highlighted areas which are not covered by existing guidance to developers. A model for how PHRs may be useful as an education tool in clinical practice was devised. Asking patients what they thought about a glaucoma PHR raised challenging questions and adds perspective to predominantly clinician led development. Listening and responding to such viewpoints is fundamental to developing more patient centred PHRs which may act both as health record and self-care educational tool to promote more holistic, efficient glaucoma care.


Pamidronate: an unrecognized problem in gastrointestinal tolerability.
Pamidronate (aminohydroxypropylidine bisphosphonate, APD) is an effective agent for treatment of Paget's disease of bone, and it has also been thought to be effective for treatment of osteoporosis. We desired to study a newer, time-release preparation of pamidronate, and carried out a placebo-controlled, double-masked study of postmenopausal osteoporosis. The original formulation was in a rapidly dissolving gelatin capsule. We encountered four episodes of esophagitis in 49 enrolled patients. We therefore discontinued treatment with this preparation and later began the study again using a standard tablet preparation. We encountered an additional case of erosive esophagitis in 1 patient of 40 receiving this tablet preparation. No patient was receiving concomitant medication which could cause esophagitis. Two of the patients gave a past history of hiatal hernia and 1 gave a history of gastric ulcer 27 years previously. The diagnosis of esophagitis was confirmed in all cases by endoscopy. Healing of the esophagitis promptly ensued after discontinuation of the pamidronate and the use of antacid medication.


MO-F-BRA-01: A Biomechanical Constraint for Intensity-Driven Deformable Alignment of Skeletal Components in the Head and Neck Region.
To introduce a biomechanical constraint into an intensity-based deformable image registration (DIR) method in order to limit nonphysical deformations of skeletal components in the neck region. On the reference image, vertebral bodies were segmented. A penalty term, based on the differences in squared inter-voxel distances within each vertebra before and after deformation, was introduced into a routinely used (ITK) intensity-based B-spline alignment algorithm. To assess accuracy, deformable image registration was performed on five pairs of cone-beam CT scans of a head and neck cancer patient. Surface registrations of individual vertebrae established their true displacements (translations and rotations). Orthogonal Procrustes analysis of transformed points within each vertebra established the estimated rotations and translations from the resultant deformation vector fields with and without the penalty term. The registration errors across all points within the vertebrae with the penalty term (0.2±0.2, 0.2±0.2, 0.3±0.2) [mm] were significantly lower than without (2.8±2.6, 3.2±2.9, 2.8±3.0) [mm], indicating that employing the penalty term successfully restricted local deformation in the region of the cervical vertebrae. The errors of the bulk translations and rotations of individual vertebrae were similarly reduced: (0.7±0.4, 0.9±0.7, 0.5±0.4) to (O.1±0.1, 0.l+0.1, 0.2±0.2) [mm] for translation and (3.4±2.6, 1.3±1.1, 1.4±1.1) to (0.7±0.6, 0.3±0.2, 0.3±0.3) [°] for rotation. The introduction of a local rigidity penalty improved the integrity of skeletal alignment under neck articulation. Further research will explore biomechanical penalties that will more realistically constrain the changes of other tissues (e.g. muscles) in the neck region. Supported by NIHR01CA59827.


Production and detection of atomic hexadecapole at Earth's magnetic field.
Optical magnetometers measure magnetic fields with extremely high precision and without cryogenics. However, at geomagnetic fields, important for applications from landmine removal to archaeology, they suffer from nonlinear Zeeman splitting, leading to systematic dependence on sensor orientation. We present experimental results on a method of eliminating this systematic error, using the hexadecapole atomic polarization moment. In particular, we demonstrate selective production of the atomic hexadecapole moment at Earth's magnetic field and verify its immunity to nonlinear Zeeman splitting. This technique promises to eliminate directional errors in all-optical atomic magnetometers, potentially improving their measurement accuracy by several orders of magnitude.


Altered neural dynamics in occipital cortices serving visual-spatial processing in heavy alcohol users.
Visual-spatial processing deficits have been previously linked to heavy alcohol use, but the underlying neurological mechanisms are poorly understood. Neuroimaging studies have shown alcohol-related aberrations in occipital cortices that appear to be associated with these neuropsychological deficits in visual-spatial processing, however the neural dynamics underlying this altered processing remains unknown. Twenty-three adults with high scores on the Alcohol Use Disorders Identification Test - Consumption (male: ⩾5, female: ⩾4) were compared to 30 demographically-matched controls with low Alcohol Use Disorders Identification Test - Consumption scores (⩽2). All participants completed a visual-spatial processing task while undergoing high-density magnetoencephalography. Time-frequency windows of interest were determined using a data-driven method, and spectrally-specific neural activity was imaged using a beamforming approach. Permutation testing of peak voxel time series was then used to statistically compare across groups. Participants with heavy alcohol use responded slower on the task and their performance was more variable. The magnetoencephalography data indicated strong theta (4-8 Hz), alpha (10-16 Hz), and gamma (62-72 Hz) responses in posterior brain regions across both groups. Following voxel time-series extraction, significant group differences were found in the left and right visual association cortices from about 375-550 ms post-stimulus, such that adults with heavy alcohol use had blunted alpha responses compared to controls. Individuals with heavy alcohol use exhibited aberrant occipital alpha activity during visual-spatial processing. These data are the first to show spectrally-specific differences during visual-spatial processing related to heavy alcohol use, and highlight alcohol's effect on systems-level neural activity.


Association of high myopia with crystallin beta A4 (CRYBA4) gene polymorphisms in the linkage-identified MYP6 locus.
Myopia is the most common ocular disorder worldwide and imposes tremendous burden on the society. It is a complex disease. The MYP6 locus at 22 q12 is of particular interest because many studies have detected linkage signals at this interval. The MYP6 locus is likely to contain susceptibility gene(s) for myopia, but none has yet been identified. Two independent subject groups of southern Chinese in Hong Kong participated in the study an initial study using a discovery sample set of 342 cases and 342 controls, and a follow-up study using a replication sample set of 316 cases and 313 controls. Cases with high myopia were defined by spherical equivalent ≤ -8 dioptres and emmetropic controls by spherical equivalent within ±1.00 dioptre for both eyes. Manual candidate gene selection from the MYP6 locus was supported by objective in silico prioritization. DNA samples of discovery sample set were genotyped for 178 tagging single nucleotide polymorphisms (SNPs) from 26 genes. For replication, 25 SNPs (tagging or located at predicted transcription factor or microRNA binding sites) from 4 genes were subsequently examined using the replication sample set. Fisher P value was calculated for all SNPs and overall association results were summarized by meta-analysis. Based on initial and replication studies, rs2009066 located in the crystallin beta A4 (CRYBA4) gene was identified to be the most significantly associated with high myopia (initial study: P = 0.02; replication study: P = 1.88e-4; meta-analysis: P = 1.54e-5) among all the SNPs tested. The association result survived correction for multiple comparisons. Under the allelic genetic model for the combined sample set, the odds ratio of the minor allele G was 1.41 (95% confidence intervals, 1.21-1.64). A novel susceptibility gene (CRYBA4) was discovered for high myopia. Our study also signified the potential importance of appropriate gene prioritization in candidate selection.


SepM, a Streptococcal Protease Involved in Quorum Sensing, Displays Strict Substrate Specificity.
Streptococcus mutans, a causative agent of dental caries, relies on multiple quorum-sensing (QS) pathways that coordinate the expression of factors needed for colonization in the oral cavity. S. mutans uses small peptides as QS signaling molecules that typically are secreted into the outside milieu. Competence-stimulating peptide (CSP) is one such QS signaling molecule that functions through the ComDE two-component signal transduction pathway. CSP is secreted through NlmTE, a dedicated ABC transporter that cleaves off the N-terminal leader peptide to generate a mature peptide that is 21 residues long (CSP-21). We recently identified a surface-localized protease, SepM, which further cleaves the CSP-21 peptide at the C-terminal end and removes the last 3 residues to generate CSP-18. CSP-18 is the active QS molecule that interacts with the ComD sensor kinase to activate the QS pathway. In this study, we show that SepM specifically cleaves CSP-21 between the Ala18 and Leu19 residues. We also show that SepM recognizes only Ala at position 18 and Leu at position 19, although some CSP-18 variants with a substitution at position 18 can function equally as well as the QS peptide. Furthermore, we demonstrate that SepM homologs from other streptococci are capable of processing CSP-21 to generate functional CSP-18. SepM is a membrane-associated streptococcal protease that processes competence-stimulating peptide (CSP) to generate an active quorum-sensing molecule in S. mutans. SepM belongs to the S16 family of serine proteases, and in this study, we found that SepM behaves as an endopeptidase. SepM displays strict substrate specificity and cleaves the peptide bond between the Ala and Leu residues. This is the first report of an endopeptidase that specifically cleaves these two residues.


Regional cerebral blood flow during alcohol withdrawal.
Serial bilateral measurements of regional cerebral blood flow (rCBF) by the xenon Xe 133 inhalation method were made during 13 withdrawal periods in 12 male alcoholics with pronounced physical dependence. A significant global reduction of rCBF was found during the first two days of withdrawal. The largest decreases were seen in cases with clouded sensorium and with an extended preceding drinking period. An rCBF distribution with relatively high temporal and low parietal flows during the first two days was coupled to aggravated symptoms. In two men who reported auditory and visual hallucinations during the measurements, elevated rCBF values were found in the temporal, Sylvian, and occipital regions.


Statins prevent pulsatile stretch-induced proliferation of human saphenous vein smooth muscle cells via inhibition of Rho/Rho-kinase pathway.
Pulsatile forces regulate vascular remodeling and trigger vascular diseases such as saphenous vein graft disease. The saphenous vein is exposed to high pressure and pulsatility only after implantation. Statins have been proved to reduce the incidence of vein graft failure. Thus, we investigated the molecular mechanisms of pulsatile stretch-induced saphenous vein smooth muscle cell (SMC) proliferation and potential beneficial effects of statins. Human saphenous vein SMCs were subjected to cyclic stretch (60 cycles/min) in Flex I plates. Cerivastatin and simvastatin significantly prevented stretch-induced increase in SMC proliferation. Stretch induced the membrane accumulation of Rho A and Rho kinase inhibitors (Y-27632 and hydroxyfasudil) and dominant negative Rho A mutant significantly prevented stretch-induced SMC proliferation. In addition, stretch increased the levels of both p44/42 mitogen-activated protein (MAP) kinase and Akt phosphorylation. MAP kinase kinase (MEK)1/2 inhibitor U0126, phosphatidylinositol (PI) 3-kinase inhibitors (wortmannin and LY294002), and dominant negative Akt mutant significantly prevented stretch-induced SMC proliferation. Cerivastatin significantly prevented stretch-induced membrane accumulation of Rho A. On the other hand, stretch-induced phosphorylation of p44/42 MAP kinase and Akt was not prevented by cerivastatin. Mevalonate restored the preventive effect of cerivasatain on stretch-induced Rho A membrane accumulation. Stretch induced hyperphosphorylation of retinoblastoma protein (pRb), which was prevented by cerivastatin and the Rho kinase inhibitors. Statins prevent stretch-induced saphenous vein SMC proliferation via inhibition of the Rho/Rho-kinase pathway. This may explain the beneficial effects of this class of drug, especially for patients after coronary artery bypass grafting.


Analysis and modeling for fiber-optic gyroscope scale factor based on environment temperature.
To explore and reduce the nonlinear error and temperature dependency of fiber-optic gyroscope (FOG) scale factor, a scale factor modeling method based on temperature is presented in this paper. A hyperbolic curve fitting is proposed according to the characteristic of scale factor under stable temperature at first. Compared to traditional modeling methods, it shows that a higher precision model of scale factor can be obtained. Then the influence of temperature on scale factor is analyzed and then the hyperbolic curve fitting method is extended based on temperature, making it possible to work over the whole potential temperature range of the FOG without degrading the performance. This paper also provides the experimental and verification results. It can be seen that a high precision model of scale factor has been established, the temperature dependency of scale factor has been reduced effectively, and the error due to environment temperature is reduced by one order at least.


Making sense of the global health crisis: policy narratives, conflict, and global health governance.
Health has become a policy issue of global concern. Worried that the unstructured, polycentric, and pluralist nature of global health governance is undermining the ability to serve emergent global public health interests, some commentators are calling for a more systematic institutional response to the "global health crisis." Yet global health is a complex and uncertain policy issue. This article uses narrative analysis to explore how actors deal with these complexities and how uncertainties affect global health governance. By comparing three narratives in terms of their basic assumptions, the way they define problems as well as the solutions they propose, the analysis shows how the unstructured pluralism of global health policy making creates a wide scope of policy conflict over the global health crisis. This wide scope of conflict enables effective policy-oriented learning about global health issues. The article also shows how exclusionary patterns of cooperation and competition are emerging in health policy making at the global level. These patterns threaten effective learning by risking both polarization of the policy debate and unanticipated consequences of health policy. Avoiding these pitfalls, the analysis suggests, means creating global health governance regimes that promote openness and responsiveness in deliberation about the global health crisis.


Optimal procedure planning and guidance system for peripheral bronchoscopy.
With the development of multidetector computed-tomography (MDCT) scanners and ultrathin bronchoscopes, the use of bronchoscopy for diagnosing peripheral lung-cancer nodules is becoming a viable option. The work flow for assessing lung cancer consists of two phases: 1) 3-D MDCT analysis and 2) live bronchoscopy. Unfortunately, the yield rates for peripheral bronchoscopy have been reported to be as low as 14%, and bronchoscopy performance varies considerably between physicians. Recently, proposed image-guided systems have shown promise for assisting with peripheral bronchoscopy. Yet, MDCT-based route planning to target sites has relied on tedious error-prone techniques. In addition, route planning tends not to incorporate known anatomical, device, and procedural constraints that impact a feasible route. Finally, existing systems do not effectively integrate MDCT-derived route information into the live guidance process. We propose a system that incorporates an automatic optimal route-planning method, which integrates known route constraints. Furthermore, our system offers a natural translation of the MDCT-based route plan into the live guidance strategy via MDCT/video data fusion. An image-based study demonstrates the route-planning method's functionality. Next, we present a prospective lung-cancer patient study in which our system achieved a successful navigation rate of 91% to target sites. Furthermore, when compared to a competing commercial system, our system enabled bronchoscopy over two airways deeper into the airway-tree periphery with a sample time that was nearly 2 min shorter on average. Finally, our system's ability to almost perfectly predict the depth of a bronchoscope's navigable route in advance represents a substantial benefit of optimal route planning.


Atomic structure of a folate/FAD-dependent tRNA T54 methyltransferase.
tRNAs from all 3 phylogenetic domains have a 5-methyluridine at position 54 (T54) in the T-loop. The methyl group is transferred from S-adenosylmethionine by TrmA methyltransferase in most Gram-negative bacteria and some archaea and eukaryotes, whereas it is transferred from 5,10-methylenetetrahydrofolate (MTHF) by TrmFO, a folate/FAD-dependent methyltransferase, in most Gram-positive bacteria and some Gram-negative bacteria. However, the catalytic mechanism remains unclear, because the crystal structure of TrmFO has not been solved. Here, we report the crystal structures of Thermus thermophilus TrmFO in its free form, tetrahydrofolate (THF)-bound form, and glutathione-bound form at 2.1-, 1.6-, and 1.05-A resolutions, respectively. TrmFO consists of an FAD-binding domain and an insertion domain, which both share structural similarity with those of GidA, an enzyme involved in the 5-carboxymethylaminomethylation of U34 of some tRNAs. However, the overall structures of TrmFO and GidA are basically different because of their distinct domain orientations, which are consistent with their respective functional specificities. In the THF complex, the pteridin ring of THF is sandwiched between the flavin ring of FAD and the imidazole ring of a His residue. This structure provides a snapshot of the folate/FAD-dependent methyl transfer, suggesting that the transferring methylene group of MTHF is located close to the redox-active N5 atom of FAD. Furthermore, we established an in vitro system to measure the methylation activity. Our TrmFO-tRNA docking model, in combination with mutational analyses, suggests a catalytic mechanism, in which the methylene of MTHF is directly transferred onto U54, and then the exocyclic methylene of U54 is reduced by FADH(2).


On the kinetics of suicide substrates.
We consider a realistic suicide substrate reaction which can be represented by four rate equations for the concentrations of the various molecules as functions of time. We present a general procedure to obtain accurate, approximate solutions analytically in terms of the rate equation parameters. This systematic technique provides more accurate approximations to the exact (numerical) solutions than other approximate methods which have been proposed based on a pseudo-steady state hypothesis.


Radiation resistance in breast cancer: are CD44+/CD24-/proteosome low/PKH26+ cells to blame?
Identification and characterization of cancer-initiating cells (CICs) enriched for stem cell-like functions and the establishment of a link between CICs and tumor recurrence, chemotherapy resistance and radiation resistance, and metastasis have been the focus of cancer research for the last eight years. Although this field has its share of controversies, it is becoming apparent that cells isolated from recurrent or residual tumors or both are enriched for cancer cells that have a specific phenotype compared with heterogeneous cells in the primary tumor. Enrichment of CICs in tumors subjected to radiation therapy could be due in part to the delivery of sublethal doses of treatment and the efficient radical scavenging system within CICs. Sublethal doses of radiation are sufficient to induce senescence of non-CICs while forcing CICs to gain several new properties related to cell cycle progression in addition to maintaining or enhancing stem cell characteristics of pre-treatment CICs. Characterizing pathways responsible for the increase in CICs after therapy and exploiting the unique characteristics of therapy-resistant CICs for developing targeted therapies are becoming a central focus of research in the rapidly evolving field of CICs.


Dissecting the Heterogeneous Population Genetic Structure of Candida albicans: Limitations and Constraints of the Multilocus Sequence Typing Scheme.
Candida albicans is a fungal opportunistic pathogen of significant public health importance mainly due to the recent emergence of strains with increased aggressiveness and antifungal resistance. Here, we aimed to describe the epidemiological profiles and approximate the population structure of C. albicans by analyzing the C. albicans multilocus sequence typing (MLST) database (Calb-MLST-DB), which contains the largest publically available dataset for this species. Based on 4,318 database isolates, we confirmed the ubiquitous nature of C. albicans including a group of diploid sequence types (DSTs) obtained from Healthy individuals exclusively (taken as an indicator of lack of association with illnesses in its host), until isolates established from Non-Healthy individuals (potentially associated with pathogenic processes) and other DSTs reported in both types (Healthy and Non-Healthy). The highest number of reported DSTs was related to blood, oral and vaginal swabs (32.4, 20.5, and 13.8%, respectively). High genetic diversity was observed in the seven housekeeping genes included in the MLST scheme, with a diverse population structure (154 clonal complexes, CCs; and a high number of singletons, n = 1,074). Phylogenetic reconstruction on the concatenated alignment of these housekeeping genes for all the reported DSTs (n = 3,483) was partially concordant with the CC assignment, however, an absence of bootstrap threshold supported nodes or p-distance, and the lack of association with the other epidemiological variables, evidenced the limitations of the MLST scheme. Marked genetic admixture signals were identified by STRUCTURE, with the majority being attributable to recombination events according to the RDP program results, although another type of exchange event cannot be ruled out. Our results reaffirm the genetic diversity inherent in the genes used for the MLST scheme, which are associated with the chromosomal remodeling already proposed for C. albicans. This was also corroborated with an internal validation at a micro geographical scale. Despite these results are biased due to the unavailability of considering the broad global spectrum of C. albicans isolates around the world. This suggests that the strategy used to population type this pathogen should be reevaluated to improve epidemiological monitoring of its health impact.


Partial artificial heart (ALVAD) use with subsequent cardiac and renal allografting in a patient with stone heart syndrome.
The abdominal left ventricular assist device (ALVAD) is an order of magnitude more effective than conventional intra-aortic balloon pumping (IABP) in unloading and providing circulatory support to the failing left ventricle. This is a report of a unique case which demonstrates that in the absence of pulmonary vascular obstruction or constriction, the ALVAD can substitute for both left and right heart function. A 21-year-old patient with a congenital bicuspid aortic valve developed acute valvular endocarditis which rapidly progressed to congestive heart failure. An operation was undertaken, the mitral and aortic valves were excised and replaced by porcine heterografts, and a fistula from the right sinus of Valsalva to the right ventricle was closed. When coronary circulation was restored, irreversible ischemic contracture of the left ventricle, or "stone heart" syndrome, developed and emergency ALVAD or partial artificial heart implantation was effected. This device functioned as a total artificial heart for nearly six days, while a donor heart was sought. The patient then underwent removal of the ALVAD and cardiac and renal allografting. The transplanted heart functioned well, but the patient expired fifteen days later from gram-negative sepsis.


Astragaloside IV alleviates hypoxia/reoxygenation-induced neonatal rat cardiomyocyte injury via the protein kinase a pathway.
Astragaloside IV (As-IV) exerts beneficial effects on hypoxia/reoxygenation (H/R)-induced cardiomyocyte injury, possibly through normalization of sarcoplasmic reticulum Ca(2+) ATPase (SERCA2a) function. The exact mechanisms remain unknown. This study was designed to investigate the role of protein kinase A (PKA) in the protective effect of As-IV on SERCA2a function. Cultured cardiomyocytes from neonatal rats were exposed to 6 h of hypoxia followed by 3 h of reoxygenation (H/R) with or without As-IV treatment. Myocyte injury was determined by the creatine kinase (CK)-MB fraction in supernatant. Myocardial SERCA2a activity and PKA kinase activity were assessed. PKA subunit mRNA expression and Ser(16) phosphorylated phospholamban (Ser(16)-PLN) protein expression were detected by real-time PCR and Western blot, respectively. The administration of As-IV significantly decreased CK-MB release and restored SERCA2a activity in H/R cardiomyocytes. The mRNAs of PKA subunits, PKA-RIα, PKA-RIIα, PKA-RIIβ, PKA-Cα and PKA-Cβ, were downregulated in H/R cardiomyocytes. However, PKA-Cα mRNA expression was significantly increased after As-IV treatment. Meanwhile, there was a tendency to recovery of the H/R-induced PKA kinase activity decrease after As-IV treatment. The expression of Ser(16)-PLN protein, which is specifically phosphorylated by PKA, was upregulated in As-IV-treated H/R cardiomyocytes. These results suggest that the cardioprotection of As-IV may be through the upregulation of PKA and Ser(16)-PLN, thereby restoring SERCA2a function in H/R injury.


Uptake of community-based HIV testing during a multi-disease health campaign in rural Uganda.
The high burden of undiagnosed HIV in sub-Saharan Africa is a major obstacle for HIV prevention and treatment. Multi-disease, community health campaigns (CHCs) offering HIV testing are a successful approach to rapidly increase HIV testing rates and identify undiagnosed HIV. However, a greater understanding of population-level uptake is needed to maximize effectiveness of this approach. After community sensitization and a census, a five-day campaign was performed in May 2012 in a rural Ugandan community. The census enumerated all residents, capturing demographics, household location, and fingerprint biometrics. The CHC included point-of-care screening for HIV, malaria, TB, hypertension and diabetes. Residents who attended vs. did not attend the CHC were compared to determine predictors of participation. Over 12 days, 18 census workers enumerated 6,343 residents. 501 additional residents were identified at the campaign, for a total community population of 6,844. 4,323 (63%) residents and 556 non-residents attended the campaign. HIV tests were performed in 4,795/4,879 (98.3%) participants; 1,836 (38%) reported no prior HIV testing. Of 2674 adults tested, 257 (10%) were HIV-infected; 125/257 (49%) reported newly diagnosed HIV. In unadjusted analyses, adult resident campaign non-participation was associated with male sex (62% male vs. 67% female participation, p = 0.003), younger median age (27 years in non-participants vs. 32 in participants; p<0.001), and marital status (48% single vs. 71% married/widowed/divorced participation; p<0.001). In multivariate analysis, single adults were significantly less likely to attend the campaign than non-single adults (relative risk [RR]: 0.63 [95% CI: 0.53-0.74]; p<0.001), and adults at home vs. not home during census activities were significantly more likely to attend the campaign (RR: 1.20 [95% CI: 1.13-1.28]; p<0.001). CHCs provide a rapid approach to testing a majority of residents for HIV in rural African settings. However, complementary strategies are still needed to engage young, single adults and achieve universal testing.


Calcium supplements controversy in osteoporosis: a physiological mechanism supporting cardiovascular adverse effects.
Recently, administration of calcium supplements for the prevention and treatment of osteoporosis has become a highly controversial issue. The findings of epidemiological studies are not necessarily supportive of the practice and are also open to different interpretations. In this article, we attempt to broaden the discussion and provide evidence that calcium supplementation may fail to compensate for renal calcium loss, and also that the resultant increased calcium load in the circulation could lead to extraskeletal deposition, including in the coronary arteries.


Length of negative resection margin does not affect local recurrence and survival in the patients with gastric cancer.
To investigate the influence of the resection margin on local recurrence and survival in gastric cancer patients. We reviewed the medical records of 1788 patients who had undergone gastrectomy for gastric cancer at the Seoul National University Bundang Hospital, South Korea, between May 2003 and July 2009. The patients were divided into early and advanced gastric cancer groups. In each group, we analyzed the relationship between clinicopathologic factors and survival outcomes, and compared the hazard rates of event occurrence between patients with resection margins above and below the cut-off value, using a Cox proportional hazard model. The early and advanced gastric cancer groups included 1001 and 787 patients, respectively. The hazard rates of event occurrence did not significantly differ between the patients with resection margins above the cut-off value and those with resection margins below the cut-off value (P > 0.05, in all comparisons). Based on the multivariable analyses, the proximal and distal resection margins were not significantly associated with survival outcomes and local recurrence (P > 0.05, in all analyses). The proximal or distal resection margins did not affect the prognosis of patients with gastric cancer if the margins were pathologically negative.


Hikikomori : Multidimensional understanding, assessment, and future international perspectives.
Hikikomori, a severe form of social withdrawal, has long been observed in Japan mainly among youth and adolescents since around the 1970s, and has been especially highlighted since the late 1990s. Moreover, hikikomori-like cases have recently been reported in many other countries. Hikikomori negatively influences not only the individual's mental health and social participation, but also wider education and workforce stability, and as such is a novel urgent global issue. In this review, we introduce the history, definition, diagnostic evaluation, and interventions for hikikomori and also the international prevalence of hikikomori outside Japan. We propose a hypothesis regarding the globalization of hikikomori based on domestic and international perspectives. In addition, we introduce our latest assessment system for hikikomori (including the latest version of the 'proposed diagnostic criteria of hikikomori for the future DSM/ICD diagnostic systems') and propose therapeutic strategies, including family approaches and individualized therapies. Finally, we present future challenges that may lead to solutions for an internationalized hikikomori.


Transplant tourism or international transplant medicine? A case for making the distinction.
Transplant tourism is routinely denounced by influential voices such as the World Health Organization, the Declaration of Istanbul and the Madrid Resolution as an unethical solution to worldwide organ shortages. Instead, it is suggested that national deceased donor schemes and multinational organ-sharing programs are the only acceptable avenues for addressing the organ shortage crisis. The present demand for self-sufficiency in organ supply responds to risks such as poor clinical outcomes, and exploitation of the poor through the various commercial practices of transplant tourism. However, opponents of transplant tourism say little about what governments should do to ensure that their citizens have real and comprehensive access to all forms of transplantation. To address this complex question, we describe a current practice of international transplant medicine in Singapore. It addresses salient concerns with transplant tourism and supports the principle of national self-sufficiency in organ supply, even as its health care system thrives and expands comprehensive transplant services to its citizens by catering to international patients. We offer a critical appraisal of the Singaporean system, and some suggestions to minimize the risk of abuse by international patients or operatives of illegal organ markets.


Resistance to Thyroid Hormone Alpha in an 18-Month-Old Girl: Clinical, Therapeutic, and Molecular Characteristics.
Recently, the first patients with resistance to thyroid hormone alpha (RTHα) due to inactivating mutations in the thyroid hormone receptor alpha (TRα) were identified. These patients are characterized by growth retardation, variable motor and cognitive defects, macrocephaly, and abnormal thyroid function tests. The objective was to characterize a young girl (18 months old) with a mutation in both TRα1 and TRα2, and to study the effects of early levothyroxine (LT4) treatment. The patient was assessed clinically and biochemically before and during 12 months of LT4 treatment. In addition, the consequences of the mutation for TRα1/2 receptor function were studied in vitro. At 18 months of age, the patient presented with axial hypotonia, delayed motor development, severe growth retardation, and abnormally elevated triiodothyronine (T3)/thyroxine (T4) ratios. RTHα was suspected, and concomitantly a c.632A>G/p.D211G missense mutation was identified, affecting both the TRα1 and TRα2 proteins. This mutation was also found in the girl's father. LT4 treatment was started, resulting in a marked improvement of her hypotonia, motor skills, and growth. Functionally, the missense mutation led to decreased transcriptional activity of TRα1, which could be overcome by higher T3 levels in vitro. The mutant TRα1 showed a moderate dominant negative activity on wild type (WT) TRα1. In contrast, WT TRα2 and mutant TRα2 had negligible transcriptional activity and showed no dominant-negative effect over TRα1. This report describes the phenotype of a young RTHα patient with a mild TRα mutation before and during early LT4 treatment. Treatment had beneficial effects on her muscle tone, motor development, and growth.


Literature-based condition-specific miRNA-mRNA target prediction.
miRNAs are small non-coding RNAs that regulate gene expression by binding to the 3'-UTR of genes. Many recent studies have reported that miRNAs play important biological roles by regulating specific mRNAs or genes. Many sequence-based target prediction algorithms have been developed to predict miRNA targets. However, these methods are not designed for condition-specific target predictions and produce many false positives; thus, expression-based target prediction algorithms have been developed for condition-specific target predictions. A typical strategy to utilize expression data is to leverage the negative control roles of miRNAs on genes. To control false positives, a stringent cutoff value is typically set, but in this case, these methods tend to reject many true target relationships, i.e., false negatives. To overcome these limitations, additional information should be utilized. The literature is probably the best resource that we can utilize. Recent literature mining systems compile millions of articles with experiments designed for specific biological questions, and the systems provide a function to search for specific information. To utilize the literature information, we used a literature mining system, BEST, that automatically extracts information from the literature in PubMed and that allows the user to perform searches of the literature with any English words. By integrating omics data analysis methods and BEST, we developed Context-MMIA, a miRNA-mRNA target prediction method that combines expression data analysis results and the literature information extracted based on the user-specified context. In the pathway enrichment analysis using genes included in the top 200 miRNA-targets, Context-MMIA outperformed the four existing target prediction methods that we tested. In another test on whether prediction methods can re-produce experimentally validated target relationships, Context-MMIA outperformed the four existing target prediction methods. In summary, Context-MMIA allows the user to specify a context of the experimental data to predict miRNA targets, and we believe that Context-MMIA is very useful for predicting condition-specific miRNA targets.


In vitro evaluation of dendrimer-polymer hybrid nanoparticles on their controlled cellular targeting kinetics.
Although polymeric nanoparticles (NPs) and dendrimers represent some of the most promising cancer-targeting nanocarriers, each of them has drawbacks such as limited tissue diffusivity/tumor penetration and rapid in vivo elimination, respectively. To address these issues, we have designed a multiscale hybrid NP system (nanohybrid) that combines folate (FA)-targeted poly(amidoamine) dendrimers and poly(ethylene glycol)-b-poly(d,l-lactide) NPs. The nanohybrids (∼100 nm NPs encapsulating ∼5 nm targeted dendrimers) were extensively characterized through a series of in vitro experiments that validate the design rationale of the system, in an aim to simulate their in vivo behaviors. Cellular uptake studies using FA receptor (FR)-overexpressing KB cells (KB FR(+)) revealed that the nanohybrids maintained high FR selectivity resembling the selectivity of free dendrimers, while displaying temporally controlled cellular interactions due to the presence of the polymeric NP shells. The cellular interactions of the nanohybrids were clathrin-dependent (characteristic of polymer NPs) at early incubation time points (4 h), which were partially converted to caveolae-mediated internalization (characteristic of FA-targeted dendrimers) at longer incubation hours (24 h). Simulated penetration assays using multicellular tumor spheroids of KB FR(+) cells also revealed that the targeted dendrimers penetrated deep into the spheroids upon their release from the nanohybrids, whereas the NP shell did not. Additionally, methotrexate-containing systems showed the selective, controlled cytotoxicity kinetics of the nanohybrids. These results all demonstrate that our nanohybrids successfully integrate the unique characteristics of dendrimers (effective targeting and penetration) and polymeric NPs (controlled release and suitable size for long circulation) in a kinetically controlled manner.


Methylphenidate and the risk of burn injury among children with attention-deficit/hyperactivity disorder.
Attention-deficit/hyperactivity disorder (ADHD) is associated with a higher risk of burn injury than in the normal population. Nevertheless, the influence of methylphenidate (MPH) on the risk of burn injury remains unclear. This retrospective cohort study analysed the effect of MPH on the risk of burn injury in children with ADHD. Data were from Taiwan's National Health Insurance Research Database (NHIRD). The sample comprised individuals younger than 18 years with a diagnosis of ADHD (n = 90 634) in Taiwan's NHIRD between January 1996 and December 2013. We examined the cumulative effect of MPH on burn injury risk using Cox proportional hazards models. We conducted a sensitivity analysis for immortal time bias using a time-dependent Cox model and within-patient comparisons using the self-controlled case series model. Children with ADHD taking MPH had a reduced risk of burn injury, with a cumulative duration of treatment dose-related effect, compared with those not taking MPH. Compared with children with ADHD not taking MPH, the adjusted hazard ratio for burn injury was 0.70 in children taking MPH for <90 days (95% confidence interval (CI) 0.64-0.77) and 0.43 in children taking MPH for ≥90 days (95% CI 0.40-0.47), with a 50.8% preventable fraction. The negative association of MPH was replicated in age-stratified analysis using time-dependent Cox regression and self-controlled case series models. This study showed that MPH treatment was associated with a lower risk of burn injury in a cumulative duration of treatment dose-related effect manner.


Efficacy of percutaneous epidural neuroplasty does not correlate with dural sac cross-sectional area in single level disc disease.
Percutaneous epidural neuroplasty (PEN) is a minimally invasive treatment. The efficacy of PEN has been relatively well investigated; however, the relationship between the clinical effectiveness of PEN and the severity of spinal canal stenosis by disc material has not yet been established. The purpose of this study was to compare clinical outcomes of PEN according to the dural sac cross-sectional area in single level disc disease. This study included 363 patients with back pain from single level disc disease with and without radiculopathy. Patients were categorized into groups according to spinal canal compromise by disc material: Category 1, less or more than 50%; and Category 2, three subgroups with lesser than a third, between a third and two thirds, and more than two thirds. Clinical outcomes were assessed according to the Visual Analog Scale (VAS) score for back pain and leg pain and Odom's criteria at 1, 3, 6, 12, and 24 months after treatment. The demographic data showed no difference between groups according to spinal canal compromise by disc material except age (older age correlated with more spinal canal compromise). The dural sac cross-sectional area did not correlate with the VAS scores for back and leg pain after PEN in single level disc disease in Groups 1 and 2. Odom's criteria after PEN were also not different according to dural sac cross-sectional area by disc material. PEN is an effective procedure in treating single level lumbar disc herniation without affecting dural sac cross-sectional area.


The early evolution of the tetrapod humerus.
A tetrapod humerus from the Late Devonian of Pennsylvania has a novel mix of primitive and derived characters. A comparative analysis of this fossil and other relevant humeri from the Devonian shows that the role of the limb in propping the body arose first in fish fins, not tetrapod limbs. The functional diversity of the earliest known limbs includes several different kinds of appendage design. This functional diversity was achieved with a humeral architecture that was remarkably conserved during the Devonian.


Myelination and motor coordination are increased in transferrin transgenic mice.
Myelin deficiency in the central nervous system (CNS) can cause severe disabling conditions. Most of the transgenic mice models overexpressing myelin components have limitations for investigators of myelin deficiency and myelin therapy as they severely alter CNS architecture. It has been postulated that transferrin (Tf) is involved in oligodendrocyte (OL) maturation and myelinogenesis. Because Tf is not an intrinsic myelin constituent, we decided to investigate if its overexpression could have an impact on the myelination process without affecting myelin integrity. We generated transgenic mice containing the complete human Tf gene specifically overexpressed in OLs. This overexpression leads to more than a 30% increase in myelin components, such as galactolipids, phospholipids, and proteins. Electron microscopy showed that myelin is structurally normal in terms of thickness and compaction. Behavior analysis showed that mice do not display significant modifications in their locomotion and cognitive and emotional abilities. Furthermore, in one of the genetic background, animals presented a significant increase in motor coordination. We did not find any modification in OL number during early postnatal development, suggesting that Tf does not act on OL proliferation. In addition, the levels of iron and ferritin remained unchanged in the brain of transgenic mice compared to control mice. Our findings indicate that, besides its known iron transport function, Tf is able to influence myelination process and induce behavioral improvements in mice.


Destructive tension: mathematics versus experience--the progress and control of the 2001 foot and mouth disease epidemic in Great Britain.
The 2001 foot and mouth disease epidemic in Great Britain was characterised by control using both traditional and novel methods, some resulting from conclusions of mathematical models. Seven days before the implementation of the novel controversial automatic pre-emptive culling of all susceptible livestock on premises adjacent to infected premises (the 'contiguous cull'), the spread of infection had already been controlled by a combination of the traditional stamping out policy with a national movement ban on livestock. A second controversial novel policy requiring the slaughter of sheep within 3 km of premises on which disease had been confirmed (the 3-km cull) also commenced after the peak of infection spread, was untargeted and took several weeks to complete; serosurveillance of culled sheep detected infection in only one flock, suggesting that cryptic infection of sheep was not propagating the epidemic. Extensive post-epidemic serological surveillance of sheep found only a small number of seropositive animals in a very few flocks, suggesting that foot and mouth disease may self-limit in extensive sheep populations. The epidemic was finally brought to an end following the introduction of enhanced agricultural movement restrictions and biosecurity measures. A welfare culling scheme of unaffected animals was required to support the prolonged national livestock movement ban. The models that supported the contiguous culling policy were severely flawed, being based on data from dissimilar epidemics; used inaccurate background population data, and contained highly improbable biological assumptions about the temporal and quantitative parameters of infection and virus emission in infected herds and flocks.


Prevalence and determinants of occupational exposures to blood and body fluids among health workers in two tertiary hospitals in Nigeria.
Healthcare associated infections among health workers commonly follow occupational exposures to pathogens infecting blood or body fluids of patients. We evaluated the prevalence and determinants of occupational exposures to blood/body fluids among health workers in two tertiary hospitals in Nigeria. In a cross section study undertaken in two tertiary hospitals in North-central and South-south Nigeria in 2011, a structured self-administered questionnaire was used to obtain demographic data and occupational exposures to blood/body fluids in the previous year from doctors, nurses and laboratory scientists. Independent predictors of occupational exposures were determined in an unconditional logistic regression model. Out of 290 health workers studied, 75.8%, 44.7%, 32.9%, 33.9% and 84.4% had skin contact with patient's blood, needle stick injuries, cut by sharps, blood/body fluid splashes to mucous membranes and one or more type of exposures respectively. Ninety one percent, 86%, 71.1%, 87.6%, 81.3%, and 84.4% of house officers, resident doctors, consultant doctors, staff nurses, principal/chief nursing officers and laboratory scientists, respectively had one or more type of exposures in the previous year (P>0.05). Professional group was found to be the only independent predictor of cut by sharps. House officers and nurses had higher and more frequent occupational exposures than other professional groups. Our results suggest high rates of occupational exposures to blood/body fluid among health workers in Nigeria, especially among newly qualified medical doctors and nurses. Health facilities in Nigeria ought to strengthen infection prevention and control practices while targeting high risk health workers such as house officers and nurses.


Bisphenol A (BPA) in China: a review of sources, environmental levels, and potential human health impacts.
Bisphenol A (BPA), identified as an endocrine disruptor, is an industrially important chemical that is used as a raw material in the manufacture of many products such as engineering plastics (e.g., epoxy resins/polycarbonate plastics), food cans (i.e., lacquer coatings), and dental composites/sealants. The demand and production capacity of BPA in China have grown rapidly. This trend will lead to much more BPA contamination in the environmental media and in the general population in China. This paper reviews the current literature concerning the pollution status of BPA in China (the mainland, Hong Kong, and Taiwan) and its potential impact on human health. Due to potential human health risks from long-term exposure to BPA, body burden of the contaminant should be monitored.


A new method for sampling African swine fever virus genome and its inactivation in environmental samples.
African swine fever (ASF) is currently the most dangerous disease for the global pig industry, causing huge economic losses, due to the lack of effective vaccine or treatment. Only the early detection of ASF virus (ASFV) and proper biosecurity measures are effective to reduce the viral expansion. One of the most widely recognized risks as regards the introduction ASFV into a country is infected animals and contaminated livestock vehicles. In order to improve ASF surveillance, we have assessed the capacity for the detection and inactivation of ASFV genome by using Dry-Sponges (3 M) pre-hydrated with a new surfactant liquid. We sampled different surfaces in ASFV-contaminated facilities, including animal skins, and the results were compared to those obtained using a traditional sampling method. The surfactant liquid successfully inactivated the virus, while ASFV DNA was well preserved for the detection. This is an effective method to systematically recover ASFV DNA from different surfaces and skin, which has a key applied relevance in surveillance of vehicles transporting live animals and greatly improves animal welfare. This method provides an important basis for the detection of ASFV genome that can be assessed without the biosafety requirements of a BSL-3 laboratory at least in ASF-affected countries, which may substantially speed up the early detection of the pathogen.


Assessment of iodine status in children, adults, pregnant women and lactating women in iodine-replete areas of China.
Iodine deficiency disorders (IDD) are widespread in China. Presently, IDD have been put under control by Universal Salt Iodisation (USI) in China; however, there is a lack of evidence on whether the iodine status in adults, pregnant women and lactating women is optimal. This study was therefore conducted to assess the iodine nutrition and thyroid function of children, adults, pregnant women and lactating women residing in areas where the USI program is fully established. Six areas were selected according to the geographical regions in China. In each of these areas, we selected 4 distinct groups of subjects (children, adults, pregnant women and lactating women) in regions where the coverage rate of iodised salt was more than 95% and the levels of iodine and fluoride in drinking water were less than or equal to 10 µg/L and 1 mg/L, respectively. We tested the iodine content of salt, urinary iodine (UI), free thyroxin (FT4), thyrotropin (TSH), thyroglobulin (Tg), thyroglobulin antibody (Tg-Ab) and antimicrosomal antibody (TM-Ab) in the 4 groups, and examined the thyroid volume in children. The median urinary iodine (MUI) concentrations were 271.4 μg/L, 260.2 μg/L, 205.9 μg/L and 193.9 μg/L in children, adults, pregnant women and lactating women, respectively; MUI in children and adults were more than adequate. The goitre prevalence (GP) in children was 6.70%. The odds ratios (OR) of subclinical hypothyroidism in the Tg-Ab- or TM-Ab-positive groups were 3.80, 7.65, 2.01 and 7.47 for children, adults, pregnant women and lactating women, respectively, compared with the negative groups. The iodine status in children and adults is above the requirement, we should reduce their iodine intake. Subclinical hypothyroidism easily occurs in the Tg-Ab or TM-Ab positive groups.


Association of FcGRIIa with Graves' disease: a potential role for dysregulated autoantibody clearance in disease onset/progression.
Although autoantibody production is a key feature of autoimmunity, it is not known whether variation in autoantibody production and clearance pathways is involved in disease susceptibility. The Fc Gamma Receptor IIa (FcGRIIa) molecule is involved in the clearance of autoantibodies and a functional single nucleotide polymorphism (SNP), rs1801274, which has been shown to alter autoantibody clearance, has been associated with a number of autoimmune diseases (AIDs) including systemic lupus erythematosus and type 1 diabetes. This study aimed to determine whether FcGRIIa is associated with Graves' disease (GD) in the UK Caucasian population by Tag SNP screening common polymorphisms within the FcGRIIa region. A case control association study investigating nine Tag SNPs within FcGRIIa, which captured the majority of known common variation within this gene region. A dataset comprising 2504 UK Caucasian GD patients and 2784 geographically matched controls taken from the 1958 British Birth cohort. We used the chi(2)-test to investigate association between the Tag SNPs and GD. Association between the rs1801274 (P = 0.003, OR = 1.12 [95% CI = 1.03-1.22] and rs6427598 (P = 0.012, OR = 0.90 [95% CI = 0.83-0.98]) SNPs and GD was observed. No other SNPs showed association with GD. No associations were seen between any of the SNPs investigated and specific GD clinical phenotypes. This study suggests that variation in FcGRIIa predisposes to GD and further supports the role of FcGRIIa as a susceptibility locus for AIDs in general.


Prolonged peritoneal gene expression using a helper-dependent adenovirus.
Encapsulating peritoneal sclerosis (EPS) is a rare complication of peritoneal dialysis. The causes of EPS are not well defined and are likely multifactorial. A suitable animal model would facilitate research into the pathophysiology and treatment of EPS. We developed a helper-dependent adenovirus that expresses both green fluorescent protein (GFP) and active transforming growth factor-beta (TGF-beta1; HDAdTGF-beta1). Mice were administered HDAdTGF-beta1 via intraperitoneal injection and the response was compared with mice administered either first-generation adenovirus expressing TGF-beta1 (AdTGF-beta1) or control adenovirus (AdGFP). HDAdTGF-beta1-treated mice continued to express the GFP reporter transgene to day 74, the end of the observation period. Transgene expression lasted less than 28 days in the animals treated with first-generation adenoviruses. Animals treated with first-generation AdTGF-beta1 demonstrated submesothelial thickening and angiogenesis at day 7, with almost complete resolution by day 28. The HDAdTGF-beta1-treated mice demonstrated progressive peritoneal fibrosis with adhesion formation and encapsulation of bowels. Weight gain was significantly reduced in animals treated with HDAdTGF-beta1 compared to both the control-treated animals and the AdTGF-beta1-treated animals. Inflammation was not a major component of the fibroproliferative response. Peritoneal administration of a first-generation AdTGF-beta1 leads to transient gene expression, resulting in a resolving fibrotic response and histology similar to that seen in simple peritoneal sclerosis. Prolonged TGF-beta1 expression induced by the helper-dependent HDAdTGF-beta1 led to changes in peritoneal morphology resembling EPS. This suggests that TGF-beta1 may be a contributing factor in both simple peritoneal sclerosis and EPS. This model will be useful for elucidation of the mechanism of EPS and evaluation of potential treatment.


Braun lipoprotein (Lpp) contributes to virulence of yersiniae: potential role of Lpp in inducing bubonic and pneumonic plague.
Yersinia pestis evolved from Y. pseudotuberculosis to become the causative agent of bubonic and pneumonic plague. We identified a homolog of the Salmonella enterica serovar Typhimurium lipoprotein (lpp) gene in Yersinia species and prepared lpp gene deletion mutants of Y. pseudotuberculosis YPIII, Y. pestis KIM/D27 (pigmentation locus minus), and Y. pestis CO92 with reduced virulence. Mice injected via the intraperitoneal route with 5 x 10(7) CFU of the Deltalpp KIM/D27 mutant survived a month, even though this would have constituted a lethal dose for the parental KIM/D27 strain. Subsequently, these Deltalpp KIM/D27-injected mice were solidly protected against an intranasally administered, highly virulent Y. pestis CO92 strain when it was given as five 50% lethal doses (LD(50)). In a parallel study with the pneumonic plague mouse model, after 72 h postinfection, the lungs of animals infected with wild-type (WT) Y. pestis CO92 and given a subinhibitory dose of levofloxacin had acute inflammation, edema, and masses of bacteria, while the lung tissue appeared essentially normal in mice inoculated with the Deltalpp mutant of CO92 and given the same dose of levofloxacin. Importantly, while WT Y. pestis CO92 could be detected in the bloodstreams and spleens of infected mice at 72 h postinfection, the Deltalpp mutant of CO92 could not be detected in those organs. Furthermore, the levels of cytokines/chemokines detected in the sera were significantly lower in animals infected with the Deltalpp mutant than in those infected with WT CO92. Additionally, the Deltalpp mutant was more rapidly killed by macrophages than was the WT CO92 strain. These data provided evidence that the Deltalpp mutants of yersiniae were significantly attenuated and could be useful tools in the development of new vaccines.


External vibration multi-directional ultrasound shearwave elastography (EVMUSE): application in liver fibrosis staging.
Shear wave speed can be used to assess tissue elasticity, which is associated with tissue health. Ultrasound shear wave elastography techniques based on measuring the propagation speed of the shear waves induced by acoustic radiation force are becoming promising alternatives to biopsy in liver fibrosis staging. However, shear waves generated by such methods are typically very weak. Therefore, the penetration may become problematic, especially for overweight or obese patients. In this study, we developed a new method called external vibration multi-directional ultrasound shearwave elastography (EVMUSE), in which external vibration from a loudspeaker was used to generate a multi-directional shear wave field. A directional filter was then applied to separate the complex shear wave field into several shear wave fields propagating in different directions. A 2-D shear wave speed map was reconstructed from each individual shear wave field, and a final 2-D shear wave speed map was constructed by compounding these individual wave speed maps. The method was validated using two homogeneous phantoms and one multi-purpose tissue-mimicking phantom. Ten patients undergoing liver magnetic resonance elastography (MRE) were also studied with EVMUSE to compare results between the two methods. Phantom results showed EVMUSE was able to quantify tissue elasticity accurately with good penetration. In vivo EVMUSE results were well correlated with MRE results, indicating the promise of using EVMUSE for liver fibrosis staging.


Preparation for the next major incident: are we ready?
In 1996, Carley and Mackway-Jones examined British hospital's readiness for a major incident. In the light of recent terrorist events in London, our group has re-visited the issue and conducted a telephone survey of relevant parties to investigate whether the situation has changed almost 10 years on. A proforma was devised, and registrars in anaesthesia, accident and emergency medicine, general surgery and trauma and orthopaedics were telephoned in trauma units across the UK and questioned about their readiness to respond to a major incident. Major incident co-ordinators for each of the units were contacted, and their planning, readiness, training opportunities, and recent rehearsals were assessed. A total of 179 registrars were contacted in 34 different units throughout Britain. One hundred and forty four responses were obtained. Sixty eight registrars (47%) had not read any of their hospitals major incident plan. Only 77 (54%) of the registrars questioned felt confident in the knowledge of their specific role during a major incident. Major incident co-ordinators were contacted at 34 hospitals, and 17 responses obtained. It was remarkably difficult to achieve even that level of response. Rehearsal of major incident plans varied widely between hospitals with 82% of hospitals having practised within the past five years but only 35% were planning for a rehearsal in the next twelve months. 25% of hospitals that responded did not hold any teaching on major incident planning at their introduction sessions for junior and middle grade doctors. Limitations to improvement of major incident planning included: lack of funds, lack of a designated full-time major incident co-ordinator, and lack of technology. There was no significant difference between units within London and those in other regions. Preparedness for major incidents in the UK remains poor 10 years after Carley and Mackway-Jones examined the issue. Effective major incident plans require forethought, organisation, briefing of relevant staff and regular rehearsal. Increased resources should be provided for this at a local level and more regular rehearsals undertaken to ensure our preparedness for future major incidents.


The relationship of nerve fibre pathology to sensory function in entrapment neuropathy.
Surprisingly little is known about the impact of entrapment neuropathy on target innervation and the relationship of nerve fibre pathology to sensory symptoms and signs. Carpal tunnel syndrome is the most common entrapment neuropathy; the aim of this study was to investigate its effect on the morphology of small unmyelinated as well as myelinated sensory axons and relate such changes to somatosensory function and clinical symptoms. Thirty patients with a clinical and electrophysiological diagnosis of carpal tunnel syndrome [17 females, mean age (standard deviation) 56.4 (15.3)] and 26 age and gender matched healthy volunteers [18 females, mean age (standard deviation) 51.0 (17.3)] participated in the study. Small and large fibre function was examined with quantitative sensory testing in the median nerve territory of the hand. Vibration and mechanical detection thresholds were significantly elevated in patients with carpal tunnel syndrome (P<0.007) confirming large fibre dysfunction and patients also presented with increased thermal detection thresholds (P<0.0001) indicative of C and Aδ-fibre dysfunction. Mechanical and thermal pain thresholds were comparable between groups (P>0.13). A skin biopsy was taken from a median nerve innervated area of the proximal phalanx of the index finger. Immunohistochemical staining for protein gene product 9.5 and myelin basic protein was used to evaluate morphological features of unmyelinated and myelinated axons. Evaluation of intraepidermal nerve fibre density showed a striking loss in patients (P<0.0001) confirming a significant compromise of small fibres. The extent of Meissner corpuscles and dermal nerve bundles were comparable between groups (P>0.07). However, patients displayed a significant increase in the percentage of elongated nodes (P<0.0001), with altered architecture of voltage-gated sodium channel distribution. Whereas neither neurophysiology nor quantitative sensory testing correlated with patients' symptoms or function deficits, the presence of elongated nodes was inversely correlated with a number of functional and symptom related scores (P<0.023). Our findings suggest that carpal tunnel syndrome does not exclusively affect large fibres but is associated with loss of function in modalities mediated by both unmyelinated and myelinated sensory axons. We also document for the first time that entrapment neuropathies lead to a clear reduction in intraepidermal nerve fibre density, which was independent of electrodiagnostic test severity. The presence of elongated nodes in the target tissue further suggests that entrapment neuropathies affect nodal structure/myelin well beyond the focal compression site. Interestingly, nodal lengthening may be an adaptive phenomenon as it inversely correlates with symptom severity.


Counseling on early childhood concerns: sleep issues, thumb sucking, picky eating, and school readiness.
Sleep issues, thumb sucking, coping with picky eating, and determining if a child is ready for school are common concerns of families with young children. Information and resources to help counsel on these topics include recommendations from the American Academy of Sleep Medicine, the American Dental Association, and the U.S. Department of Agriculture. Infant sleep times can be prolonged by unmodified or graduated extinction, maintaining routines, scheduled awakenings, and parent education. Thumb sucking can be addressed with positive reinforcement, alternative comfort measures, reminders, and child involvement in solutions. Worry about picky eating can be eased by educating parents about the dietary requirements of toddlers. Social and emotional factors most influence kindergarten success. Keeping children from starting school may not be in their best interest academically.


Improved sensitivity and specificity for resting state and task fMRI with multiband multi-echo EPI compared to multi-echo EPI at 7 T.
A multiband multi-echo (MBME) sequence is implemented and compared to a matched standard multi-echo (ME) protocol to investigate the potential improvement in sensitivity and spatial specificity at 7 T for resting state and task fMRI. ME acquisition is attractive because BOLD sensitivity is less affected by variation in T2*, and because of the potential for separating BOLD and non-BOLD signal components. MBME further reduces TR thus increasing the potential reduction in physiological noise. In this study we used FSL-FIX to clean ME and MBME resting state and task fMRI data (both 3.5mm isotropic). After noise correction, the detection of resting state networks improves with more non-artifactual independent components being observed. Additional activation clusters for task data are discovered for MBME data (increased sensitivity) whereas existing clusters become more localized for resting state (improved spatial specificity). The results obtained indicate that MBME is superior to ME at high field strengths.


Drosophila sugar receptors in sweet taste perception, olfaction, and internal nutrient sensing.
Identification of nutritious compounds is dependent on expression of specific taste receptors in appropriate taste-cell types [1]. In contrast to mammals, which rely on a single, broadly tuned heterodimeric sugar receptor [2], the Drosophila genome harbors a small subfamily of eight, closely related gustatory receptor (Gr) genes, Gr5a, Gr61a, and Gr64a-Gr64f, of which three have been proposed to mediate sweet taste [3-6]. However, expression and function of several of these putative sugar Gr genes are not known. Here, we present a comprehensive expression and functional analysis using Gr(LEXA/GAL4) alleles that were generated through homologous recombination. We show that sugar Gr genes are expressed in a combinatorial manner to yield at least eight sets of sweet-sensing neurons. Behavioral investigations show that most sugar Gr mutations affect taste responses to only a small number of sugars and that effective detection of most sugars is dependent on more than one Gr gene. Surprisingly, Gr64a, one of three Gr genes previously proposed to play a major role in sweet taste [3, 4], is not expressed in labellar taste neurons, and Gr64a mutant flies exhibit normal sugar responses elicited from the labellum. Our analysis provides a molecular rationale for distinct tuning profiles of sweet taste neurons, and it favors a model whereby all sugar Grs contribute to sweet taste. Furthermore, expression in olfactory organs and the brain implies novel roles for sugar Gr genes in olfaction and internal nutrient sensing, respectively. Thus, sugar receptors may contribute to feeding behavior via multiple sensory systems.


Neural networks for action representation: a functional magnetic-resonance imaging and dynamic causal modeling study.
Automatic mimicry is based on the tight linkage between motor and perception action representations in which internal models play a key role. Based on the anatomical connection, we hypothesized that the direct effective connectivity from the posterior superior temporal sulcus (pSTS) to the ventral premotor area (PMv) formed an inverse internal model, converting visual representation into a motor plan, and that reverse connectivity formed a forward internal model, converting the motor plan into a sensory outcome of action. To test this hypothesis, we employed dynamic causal-modeling analysis with functional magnetic-resonance imaging (fMRI). Twenty-four normal participants underwent a change-detection task involving two visually-presented balls that were either manually rotated by the investigator's right hand ("Hand") or automatically rotated. The effective connectivity from the pSTS to the PMv was enhanced by hand observation and suppressed by execution, corresponding to the inverse model. Opposite effects were observed from the PMv to the pSTS, suggesting the forward model. Additionally, both execution and hand observation commonly enhanced the effective connectivity from the pSTS to the inferior parietal lobule (IPL), the IPL to the primary sensorimotor cortex (S/M1), the PMv to the IPL, and the PMv to the S/M1. Representation of the hand action therefore was implemented in the motor system including the S/M1. During hand observation, effective connectivity toward the pSTS was suppressed whereas that toward the PMv and S/M1 was enhanced. Thus, the action-representation network acted as a dynamic feedback-control system during action observation.


Screening for reducing morbidity and mortality in malignant melanoma.
Screening for malignant melanoma has the potential to reduce morbidity and mortality from the disease through earlier detection, as prognosis is closely associated with the thickness of the lesion at the time of diagnosis. However, there are also potential harms from screening people without skin lesion concerns, such as overdiagnosis of lesions that would never have caused symptoms if they had remained undetected. Overdiagnosis results in harm through unnecessary treatment and the psychosocial consequences of being labelled with a cancer diagnosis. For any type of screening, the benefits must outweigh the harms. Screening for malignant melanoma is currently practised in many countries, and the incidence of the disease is rising sharply, while mortality remains largely unchanged. To assess the effects on morbidity and mortality of screening for malignant melanoma in the general population. We searched the following databases up to May 2018: the Cochrane Skin Specialised Register, CENTRAL, MEDLINE, Embase, and LILACS. We also searched five trials registries, checked the reference lists of included and other relevant studies for further references to randomised controlled trials (RCTs), used citation tracking (Web of Science) for key articles, and asked trialists about additional studies and study reports. RCTs, including cluster-randomised trials, of screening for malignant melanoma compared with no screening, regardless of screening modality or setting, in any type of population and in any age group where people were not suspected of having malignant melanoma. We excluded studies in people with a genetic disposition for malignant melanoma (e.g. familial atypical mole and melanoma syndrome) and studies performed exclusively in people with previous melanomas. We used standard methodological procedures expected by Cochrane. The primary outcomes of this review were total mortality, overdiagnosis of malignant melanoma, and quality of life/psychosocial consequences. We included two studies with 64,391 participants. The first study was a randomised trial of an intervention developed to increase the rate of performance of thorough skin self-examination. The intervention group received instructional materials, including cues and aids, a 14-minute instruction video, and a brief counselling session, and at three weeks a brief follow-up telephone call from a health educator, aimed at increasing performance of thorough skin self-examination. The control group received a diet intervention with similar follow-up. The trial included 1356 people, who were recruited from 11 primary care practices in the US between 2000 and 2001. Participant mean age was 53.2 years and 41.7% were men. This study did not report on any of our primary outcomes or the following secondary outcomes: mortality specific to malignant melanoma, false-positive rates (skin biopsies/excisions with benign outcome), or false-negative rates (malignant melanomas diagnosed between screening rounds and up to one year after the last round). All participants were asked to complete follow-up telephone interviews at 2, 6, and 12 months after randomisation.The second study was a pilot study for a cluster-RCT of population-based screening for malignant melanoma in Australia. This pilot trial included 63,035 adults aged over 30 years. The three-year programme involved community education, an education and support component for medical practitioners, and the provision of free skin screening services. The mean age of people attending the skin screening clinics (which were held by primary care physicians in workplaces, community venues, and local hospitals, and included day and evening sessions) was 46.5 years, and 51.5% were men. The study included whole communities, targeting participants over 30 years of age, but information on age and gender of the whole study population was not reported. Study duration was three years (1998 to 2001), and outcomes were measured at the screening clinics during these three years. There was no further follow-up for any outcomes. The control group received no programme. The ensuing, planned cluster randomised trial in 560,000 adults was never carried out due to lack of funding. At the time of this review, there are no published or unpublished data on our prespecified outcomes available, and no results for mortality outcomes from the pilot study are to be expected.The risk of bias in these studies was high for performance bias (blinding study personnel and participants) and high or unclear for detection bias (blinding of outcome assessment). Risk of bias in the other domains was either unclear or low. We were unable to assess the certainty of the evidence for our primary outcomes as planned due to lack of data. Adult general population screening for malignant melanoma is not supported or refuted by current evidence from RCTs. It therefore does not fulfil accepted criteria for implementation of population screening programmes. This review did not investigate the effects of screening people with a history of malignant melanoma or in people with a genetic disposition for malignant melanoma (e.g. familial atypical mole and melanoma syndrome). To determine the benefits and harms of screening for malignant melanoma, a rigorously conducted randomised trial is needed, which assesses overall mortality, overdiagnosis, psychosocial consequences, and resource use.


Identification of ZFR, an ancient and highly conserved murine chromosome-associated zinc finger protein.
In a screen for RNA binding proteins expressed during murine spermatogenesis, we cloned a novel, ancient zinc finger protein possessing a region common to a small class of RNA binding proteins. Zfr (zinc finger RNA binding) encodes a protein of 1052 amino acids with three widely spaced Cys2His2 zinc fingers. Outside of the zinc fingers, ZFR shares a region that is highly conserved between several RNA binding proteins containing copies of the double-stranded RNA binding motif. By northern blotting, Zfr is expressed at highest levels within the testis, ovary and brain. Immunohistochemistry and confocal microscopy were used to show that ZFR is highly expressed during meiosis I in males and females and is chromosome associated. Zfr is also expressed in Sertoli cells in the testis and granulosa cells in the ovary where it is localized to the nucleus. Using fluorescent in situ hybridization we mapped Zfr to chromosome 15 region A. ZFR appears to be an ancient protein, as apparent homologs exist in invertebrates (D. melanogaster) nematodes (C. elegans) and humans (H. sapiens).


Single molecule fluorescence resonance energy transfer scanning near-field optical microscopy: potentials and challenges.
A few years ago, single molecule Fluorescence Resonance Energy Transfer Scanning Near-Field Optical Microscope (FRET SNOM) images were demonstrated using CdSe semiconductor nanocrystal-dye molecules as donor-acceptor pairs. Corresponding experiments reveal the necessity to exploit much more photostable fluorescent centers for such an imaging technique to become a practically used tool. Here we report the results of our experiments attempting to use nitrogen vacancy (NV) color centers in nanodiamond (ND) crystals, which are claimed to be extremely photostable, for FRET SNOM. All attempts were unsuccessful, and as a plausible explanation we propose the absence (instability) of NV centers lying close enough to the ND border. We also report improvements in SNOM construction that are necessary for single molecule FRET SNOM imaging. In particular, we present the first topographical images of single strand DNA molecules obtained with fiber-based SNOM. The prospects of using rare earth ions in crystals, which are known to be extremely photostable, for single molecule FRET SNOM at room temperature and quantum informatics at liquid helium temperatures, where FRET is a coherent process, are also discussed.


Non-arteritic anterior ischaemic optic neuropathy and presumed sleep apnoea syndrome screened by the Sleep Apnea scale of the Sleep Disorders Questionnaire (SA-SDQ).
Two recent studies reported over 70% of the patients with non-arteritic anterior ischaemic optic neuropathy (NAION) had sleep apnoea syndrome (SAS) diagnosed by overnight polysomnography. The current study used the Sleep Apnea scale of the Sleep Disorders Questionnaire (SA-SDQ) to evaluate this association. A matched case-control study was conducted among 73 cases of NAION matched on age and gender to 73 controls without a history of NAION. Information regarding demographics, medical conditions, health behaviours and SAS was obtained via a telephone questionnaire that included the SA-SDQ. Conditional logistic regression was used to calculate odds ratios (OR) and the 95% confidence intervals (CI) for the association between NAION and the SA-SDQ. Cases were significantly more likely to report symptoms and characteristics consistent with SAS than controls (OR 2.62; 95% CI 1.03 to 6.60) when adjusted for medical and health behaviour characteristics. The results of this study suggest that patients with SAS are at increased risk of NAION. Additional research in a larger population is needed to confirm the observed results and validate the use of the SA-SDQ in patients with NAION.


Cardiovascular events in pregnancy with hypertrophic cardiomyopathy.
The influence of the physiological circulatory changes during pregnancy on hypertrophic cardiomyopathy (HCM) is unclear. There have been no comprehensive studies of pregnant women with HCM in the Japanese population. A total of 27 pregnancies (23 women with HCM) were retrospectively reviewed. A total of 18 cardiovascular events occurred in 13 of the 27 pregnancies (48%), and 13 of these events (76%) were related to arrhythmia. The cardiovascular events tended to occur in the early stage of pregnancy (≈30 gestational weeks) or postpartum. The events related to arrhythmia mainly occurred in the early stage of pregnancy or at approximately 30 gestational weeks. Four pregnancies were terminated because of cardiovascular events. Cardiovascular events occurred in 8 of 9 pregnancies in women on medication before pregnancy (88%), 7 of 10 pregnancies with high CARPREG score (70%), and in 9 of 12 pregnancies with high ZAHARA score (75%). Cardiovascular events occurred in more than half of the pregnant women complicated with HCM, and the arrhythmia is the most common cardiovascular event. Medication in the pre-pregnancy period, and CARPREG or ZAHARA score ≥1 were identified as risk factors of cardiac events during pregnancy or postpartum.


A novel role of human holliday junction resolvase GEN1 in the maintenance of centrosome integrity.
The maintenance of genomic stability requires accurate genome replication, repair of DNA damage, and the precise segregation of chromosomes in mitosis. GEN1 possesses Holliday junction resolvase activity in vitro and presumably functions in homology driven repair of DNA double strand breaks. However, little is currently known about the cellular functions of human GEN1. In the present study we demonstrate that GEN1 is a novel centrosome associated protein and we characterize the various phenotypes associated with GEN1 deficiency. We identify an N-terminal centrosome localization signal in GEN1, which is required and sufficient for centrosome localization. We report that GEN1 depletion results in aberrant centrosome numbers associated with the formation of multiple spindle poles in mitosis, an increased number of cells with multi-nuclei, increased apoptosis and an elevated level of spontaneous DNA damage. We find homologous recombination severely impaired in GEN1 deficient cells, suggesting that GEN1 functions as a Holliday junction resolvase in vivo as well as in vitro. Complementation of GEN1 depleted cells with various GEN1 constructs revealed that centrosome association but not catalytic activity of GEN1 is required for preventing centrosome hyper-amplification, formation of multiple mitotic spindles, and multi-nucleation. Our findings provide novel insight into the biological functions of GEN1 by uncovering an important role of GEN1 in the regulation of centrosome integrity.


Collaboration to increase colorectal cancer screening among low-income uninsured patients.
Colorectal cancer (CRC) is the second leading cause of cancer-related deaths in the United States. CRC screening allows for prevention through the removal of precancerous lesions and early detection of cancer. Ride for Life Alaska (RFL), a nonprofit organization that raises funds to fight cancer, and the Anchorage Neighborhood Health Center (ANHC), which is Alaska's largest community health center, joined efforts to provide CRC screening and outreach to an ethnically diverse group of low-income underinsured or uninsured patients residing in and around Anchorage, Alaska. RFL and ANHC worked with gastroenterologists, medical practices, and pathology services to contribute pro bono and reduced-fee services for CRC screening. Information to patients was distributed through signs in the clinic, flyers, and the ANHC website. CRC screening was increased in this population. During 2007-2009, there were 2,561 immunochemical fecal occult blood tests given to patients, and 1,558 were completed (61%); 24% were positive. Sixteen gastroenterologists, 4 medical practices, and 2 laboratories provided 111 follow-up colonoscopies and pathology services to patients identified through the CRC screening program who did not have other funding resources available for follow-up care. This program provides a model for leveraging scarce screening resources by drawing on multiple partners to increase CRC screening. Recommendations for those seeking to initiate similar programs are to have memoranda of agreement in place and a clear scope of work for all participating people and organizations to avoid delays in program implementation; hire a screening care coordinator to manage patient care and collaborate with medical practices; and identify program champions who have the energy and persistence to craft such partnerships.


Synthesis of a new class of silsesquioxanes with alkynyl and germyl functionality.
Although there are a variety of functional compounds in the vast family of silsesquioxanes (POSS), the number of protocols describing specific monosubstituted silsesquioxanes is limited. There are even fewer reports on alkynyl and/or germyl functionalities appearing together with the POSS compounds. Here, a series of novel kinds of cubic monosubstituted POSS bearing alkynyl and germyl functionality in one molecule are reported. These compounds were synthesized by effective germylative coupling of terminal alkynes with vinylgermanes in the presence of a Ru-H complex. The structures of the products were determined. Moreover, stoichiometric reactions of a complex possessing Ru-Ge were also performed to propose the general mechanistic scheme of this reaction.


Immunologic response to xenogeneic gp100 DNA in melanoma patients: comparison of particle-mediated epidermal delivery with intramuscular injection.
Prior studies show that i.m. injection of xenogeneic orthologues of melanosomal antigens (tyrosinase, gp100) induces CD8(+) T-cell responses to the syngeneic protein. To further define the optimal vaccination strategy, we conducted a pilot clinical trial comparing i.m. injection with particle-mediated epidermal delivery (PMED). Human leukocyte antigen (HLA)-A*0201(+) disease-free melanoma patients were randomized to the PMED or i.m. arm, receiving eight vaccinations over 4 months. Patients received 4 microg or 2,000 microg per injection, respectively, of mouse gp100 DNA. Peripheral blood mononuclear cells were collected, cultured with gp100 peptides, and analyzed by tetramer and intracellular cytokine staining for responses to HLA-A*0201-restricted gp100 epitopes [gp100(209-217) (ITDQVPFSV) and gp100(280-288) (YLEPGPVTA)]. Twenty-seven patients with stage IIB-IV melanoma were analyzable for immune response. The only common toxicity was grade 1 injection site reaction in nine patients with no intergroup difference, and one dose-limiting toxicity of acute hypersensitivity occurred in a PMED patient with undiagnosed gold allergy. Four of 27 patients produced gp100 tetramer(+)CD8(+) T cells, all carrying the CCR7(lo)CD45RA(lo) effector-memory phenotype. Five of 27 patients generated IFN-gamma(+)CD8(+) T cells, one who was also tetramer-positive. Overall, vaccination induced a response in 30% of patients, which was not significantly associated with study arm or clinical outcome. However, the PMED group showed a trend toward increased IFN-gamma(+)CD8(+) T-cell generation (P = 0.07). A comparable efficacy and safety profile was shown between the i.m. and PMED arms, despite a significantly decreased dose of DNA used for PMED injection.