CyclinD1, a prominent prognostic marker for endometrial diseases.
Alteration of CyclinD1 was suggested to relate with development of endometrial carcinogenesis before, however CyclinD1 expression is not well defined in endometrial hyperplasia lesions. We checked the relationship between its expression and clinic-pathological variables of endometrial lesions to explore the possibility for CyclinD1 as a potential diagnostic and prognostic marker. Cyclin D1 immunohistochemical analysis (IHC) was used to evaluate 201 fixed, paraffin-embedded endometrial samples which included simple hyperplasia (n = 27), atypical complex hyperplasia (ACH) (n = 41), endometrioid carcinoma (n = 103), endometrial serous carcinoma (ESC) (n = 21) and clear cell carcinoma (CCC) (n = 9). A breast cancer with known CyclinD1 expression was selected as a positive control in each immunohistochemistry run. We also performed follow-up study to estimate patients' prognosis. CyclinD1 was significantly overexpressed in atypical complex hyperplasia (ACH), endometrioid carcinoma and clear cell carcinoma (CCC). The positive signaling of CyclinD1 was showed less than 40% in simple hyperplasia and endometrial serous carcinoma (ESC). The high expression of CyclinD1 was observed in metastasis carcinoma group more significantly than non-metastasis carcinoma group. Kaplan Meier analysis demonstrated that patients with high CyclinD1 expression had an obviously poor prognosis than patients without CyclinD1 staining (p < 0.05). Moreover, according to multivariate Cox regression analysis, CyclinD1 expression, as crucial as metastasis, was a risk marker for overall survival rate. CyclinD1 exhibited a promising potential to predict the prognosis of patients with endometrial carcinoma. However, the statistical analysis demonstrated that CyclinD1 exhibited a poor ability to differentiate neoplastic lesions from non-neoplastic lesions; thus, the application of CyclinD1 only is not so credible for differentiation between benign and malignant lesions. The virtual slides for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/1871063048950173.


From venoms to BBB-shuttles. MiniCTX3: a molecular vector derived from scorpion venom.
The present study aims to develop chlorotoxin (CTX), from Giant Yellow Israeli scorpion venom, as a new BBB-shuttle. Minimised versions of CTX were prepared to reduce its complexity while enhancing its BBB-shuttle capacity and preserving its protease-resistance. MiniCTX3, a monocyclic lactam-bridge peptidomimetic, was capable of transporting nanoparticles across endothelial cell monolayers. Our results reveal animal venoms as an outstanding source of new families of BBB-shuttles.


The relationship between holding time and the bacterial load on surgical instruments.
The aim of this investigation was to determine the bacterial load on used instruments and to evaluate the relationship between the bacterial load and the holding time prior to cleaning. Thirty six sets were evaluated to establish the average number of bacteria per square centimeter. For the experimental study, three different bacteria were prepared in sheep blood and used to contaminate sterile stainless steel pieces with the surface of 10 cm(2). After incubation at room temperature for 2, 4, 6, 8, 12, 24, 36, and 48 h, colonies were counted and compared to time zero. Bacterial counts were between 10 and 250 CFU/cm(2), depending on the operation site. Bacterial load was found to have increased after 6 h. An increase of 3log10 CFU/cm(2) was measured after 12 h. It is imperative to clean surgical instruments in the first 6 h to ensure effective disinfection and sterility.


Human T cell activation with phytohemagglutinin. The function of IL-6 as an accessory signal.
T cell proliferation induced by the lectin PHA requires the presence of monocytes. Here, we report that IL-6 represents part of the monocyte-derived helper activity for human T cell stimulation with PHA. Resting T cells isolated from peripheral blood or tonsils and completely depleted of accessory cells (monocytes and NK cells) did not proliferate in response to PHA alone. Addition of a cell-free monocyte culture supernatant (not containing IL-2) to the PHA-stimulated cultures resulted in T cell proliferation. We identified IL-6 as an essential helper factor in these monocyte supernatants, because antisera to IL-6 (but not anti-IL-1-beta) completely neutralized the helper effect of the monocyte supernatant. Moreover, addition of purified human IL-6 (but not IL-1-beta) to T cell cultures resulted in proliferation of PHA-stimulated T cells. Although IL-1 could not restore the PHA-induced proliferative response of isolated T cells by itself, it strongly enhanced the helper effect of IL-6. In contrast to intact monocytes, IL-6 did not induce detectable IL-2 production in cultures of PHA-stimulated T cells. Furthermore, an anti-IL-2R mAb (anti-Tac) did not block the proliferative response induced by PHA and IL-6, suggesting that an IL-2-independent pathway of T cell proliferation was involved. In conclusion our results show that human IL-6, previously identified as a B cell hybridoma growth and a B cell differentiation factor, represents part of the monocyte helper function for PHA-induced human T cell activation and proliferation.


Even Worse - Risk Factors and Protective Factors for Transition from Chronic Localized Low Back Pain to Chronic Widespread Pain in General Practice: A Cohort Study.
Prospective cohort study with patients with chronic low back pain (CLBP) at primary care setting. The aim of our study was to identify predictors for transition from localized CLBP to chronic widespread pain in general practice. In contrast to the typically investigated risk factors, this study also focuses intensively on protective factors, which decrease the probability of chronic widespread pain. For this, we investigated the resources resilience and coping strategies, which are suspected as potential protective factors for incident chronic pain syndromes. In primary care, about a quarter of patients with CLBP experience chronic widespread pain (CWP). Patients experiencing localized CLBP were included and evaluated after a 6- and 12-month follow-up period regarding the development of CWP. Potential risk factors (sociodemographic data, pain characteristics, depression, anxiety, somatization), protective factors (resilience, coping strategies), and sample characteristics were assessed at baseline. Predictor identification was done by multivariate logistic regression analysis. The 1-year incidence for the onset of CWP among patients with CLBP was 23.8%. We identified the 3 risk factors, female sex, long duration of back pain, and a high rate of psychosomatic symptoms, for the onset of CWP among patients with CLBP. Coping resources and resilience had no impact on the transition from CLBP to CWP. The results suggest that CWP is no independent entity but rather a particularly negative occurrence on a continuum of chronic pain. Processes of somatization play a major role in the development of this extreme. 2.


In Vivo Performance and Properties of Tamoxifen Metabolites for CreERT2 Control.
Mutant Estrogen Receptor (ERT2) ligand-binding domain fusions with Cre recombinase are a key tool for spatio-temporally controlled genetic recombination with the Cre/lox system. CreERT2 is efficiently activated in a concentration-dependent manner by the Tamoxifen metabolite trans-4-OH-Tamoxifen (trans-4-OHT). Reproducible and efficient Cre/lox experimentation is hindered by the gradual loss of CreERT2 induction potency upon prolonged storage of dissolved trans-4-OHT, which potentially results from gradual trans-to-cis isomerization or degradation. Here, we combined zebrafish CreERT2 recombination experiments and cell culture assays to document the gradual activity loss of trans-4-OHT and describe the alternative Tamoxifen metabolite Endoxifen as more stable alternative compound. Endoxifen retains potent activation upon prolonged storage (3 months), yet consistently induces half the ERT2 domain fusion activity compared to fresh trans-4-OHT. Using 1H-NMR analysis, we reveal that trans-4-OHT isomerization is undetectable upon prolonged storage in either DMSO or Ethanol, ruling out isomer transformation as cause for the gradual loss of trans-4-OHT activity. We further establish that both trans-4-OHT and Endoxifen are insensitive to light exposure under regular laboratory handling conditions. We attribute the gradual loss of trans-4-OHT potency to precipitation over time, and show that heating of aged trans-4-OHT aliquots reinstates their CreERT2 induction potential. Our data establish Endoxifen as potent and reproducible complementary compound to 4-OHT to control ERT2 domain fusion proteins in vivo, and provide a framework for efficient chemically controlled recombination experiments.


Intraobserver and interobserver variability in schemes for estimating volume of brain lesions on MR images in multiple sclerosis.
Our goal was to evaluate the intraobserver and interobserver reproducibility of measurements of brain lesion load in multiple sclerosis (MS) by using two proposed acquisition schemes. Three-millimeter-thick conventional spin-echo (CSE) and fast fluid-attenuated inversion-recovery (FLAIR) sequences were obtained and the lesions segmented using a semiautomated technique based on local thresholding to calculate intraobserver and interobserver reproducibility. These were compared with images obtained from two separate MR units in which 5-mm CSE sequences were obtained and segmented by using the local thresholding technique and also by manual outlining. The intraobserver coefficient of variation was 4.0% (95% confidence interval [CI], 3.0% to 4.5%) for the 5-mm CSE sequence measured with manual outlining, 3.1% (95% CI, 2.5% to 3.2%) and 5.1% (95% CI, 4.1% to 5.6%) for the two sets of 5-mm CSE sequences measured using the local thresholding technique, 5.7% (95% CI, 3.9% to 6.6%) for the 3-mm CSE sequence, and 2.6% (95% CI, 2.1% to 2.7%) for the fast FLAIR sequence. The interobserver coefficient of variation was 7.1% (95% CI, 4.9% to 8.7%) and 8.3% (95% CI, 6.4% to 9.6%) for the two sets of 5-mm CSE sequences, 7.3% (95% CI, 4.7% to 9.1%) for the 3-mm CSE sequence, and 2.9% (95% CI, 2.3% to 3.3%) for the fast FLAIR sequence. The intraobserver and interobserver reproducibility of measurements obtained with the fast FLAIR technique was significantly better than those obtained with the other techniques. Our data indicate that the intraobserver and interobserver variability in quantifying MS lesions can be reduced significantly with the use of fast FLAIR sequences, while no significant improvement is gained by reducing the section thickness from 5 mm to 3 mm.


Limitation of Life-Sustaining Care in the Critically Ill: A Systematic Review of the Literature.
When life-sustaining treatments (LST) are no longer effective or consistent with patient preferences, limitations may be set so that LSTs are withdrawn or withheld from the patient. Many studies have examined the frequency of limitations of LST in intensive care unit (ICU) settings in the past 30 years. This systematic review describes variation and patient characteristics associated with limitations of LST in critically ill patients in all types of ICUs in the United States. A comprehensive search of the literature was performed by a medical librarian between December 2014 and April 2017. A total of 1,882 unique titles and abstracts were reviewed, 113 were selected for article review, and 36 studies were fully reviewed. Patient factors associated with an increased likelihood of limiting LST included white race, older age, female sex, poor preadmission functional status, multiple comorbidities, and worse illness severity score. Based on several large, multicenter studies, there was a trend toward a higher frequency of limitation of LST over time. However, there is large variability between ICUs in the proportion of patients with limitations and on the proportion of deaths preceded by a limitation. Increases in the frequency of limitations of LST over time suggests changing attitudes about aggressive end-of-life-care. Limitations are more common for patients with worse premorbid health and greater ICU illness severity. While some differences in the frequency of limitations of LST may be explained by personal factors such as race, there is unexplained wide variability between units.


Decreased Survival of Medial Pivot Designs Compared with Cruciate-retaining Designs in TKA Without Patellar Resurfacing.
The medial pivot TKA design was introduced in the 1990s. These are fixed-bearing, medial-conforming implants with virtually no translation in the medial part of the knee, in contrast to the flat lateral part of the insert allowing for translation similar to the native knee during flexion and extension. Most primary TKAs performed in Norway and Australia are cruciate-retaining. All of the medial pivot implants in our study are cruciate-sacrificing but without a post-cam mechanism. The medial pivot implant design was developed to more closely mimic native knee motion, in the hope of improving function, and not primarily as a more constrained knee for difficult cases. In the past 10 to 12 years, a second-generation medial-pivot design has emerged, but there are no larger registry studies on the survival of these implants. Both cruciate-retaining and medial pivot designs are reported in the Australian and Norwegian registries, allowing for large-scale, comparative survivorship studies. (1) Is there any difference in survival between the medial pivot design and the three most commonly used cruciate-retaining TKA designs? (2) Is there any difference in survival among the different medial pivot implant designs? (3) What are the main indications for revision of medial pivot TKAs? Registry data from the Australian Orthopaedic Association National Joint Replacement Registry and Norwegian Arthroplasty Register from 2005 until the end of 2017 were used to compare the five different brands of medial pivot TKA designs (total primary TKAs assessed: 6310). In Australia, the study group of medial pivot implants represented 9% (6012 of 72,477) of the total number of cemented/hybrid TKAs without patellar resurfacing; 345 had cementless femoral components. In Norway, the study group represented 1% (298 of 47,820) of the total number of TKAs with cemented tibias without patellar resurfacing; all had cemented femoral components. The control group consisted of the three most commonly used cruciate-retaining TKA designs (n = 70,870; Australia n = 54,554; Norway n = 16,316). All TKAs used a fixed-bearing, cemented tibial component and did not involve patella resurfacing. Kaplan-Meier survival analysis was assessed to estimate survivorship. We compared the groups by calculating the hazard ratios (HR) using Cox regression adjusted for age, gender and preoperative diagnosis with 95% CI. To answer our third question, we calculated the percentage of each revision indication from the total number of revisions in each group, and used a Cox regression analysis to compare revision causes and HRs. Analyses were performed separately by each registry. Accounting for competing risks (Fine and Gray) did not alter our findings []. After controlling for potential confounding variables such as gender, age and preoperative diagnosis, we found an increased revision risk for the medial pivot compared with cruciate-retaining TKA designs in Australia (HR 1.4 [95% CI 1.2 to 1.7]; p < 0.001), but not in Norway (HR 1.5 [95% CI 0.9 to 2.4]; p = 0.1). Two brands of the medial pivot design reported to the AOANJRR showed an increased risk of revision compared with cruciate-retaining designs: the Advance® II MP (HR 1.7 [95% CI 1.2 to 2.6]; p = 0.004) and the GMK® Sphere (HR 2.0 [95% CI 1.5 to 2.6]; p < 0.001), whereas the MRK (HR 0.7 [95% CI 0.4 to 1.5]; p = 0.4), the Evolution® MP (HR 1.4 [95% CI 1.0 to 1.9]; p = 0.06) and the SAIPH® (HR 0.9 [95% CI 0.5 to 1.5]; p = 0.7) showed no difference. The most common reasons for revision of medial pivot implants in Australia were infection (27%), pain alone (19%), patellar erosion (13%), loosening/lysis (12%); in Norway the primary indications were loosening/lysis (28%), instability (28%), malalignment (11%) and pain alone (11%). The medial pivot TKA design as a group had a higher revision rate than cruciate-retaining fixed-bearing controls in TKA performed without patellar component resurfacing. By brand, the Advance II MP and the GMK Sphere had inferior survivorship, whereas the MRK, the SAIPH and the Evolution MP had no differences in survivorship compared with cruciate-retaining controls. In Australia, TKAs with the medial pivot design without patella resurfacing had a higher rate of revisions for instability, malalignment, and patella erosion. In Norway, there was an increased risk of revision for lysis and loosening compared with the cruciate-retaining design. Several of these implants had short follow-up in this study. Further registry studies with longer follow up are therefore necessary. Level III, therapeutic study.


Considerations for improved performance of competition association assays analysed with the Motulsky-Mahan's "kinetics of competitive binding" model.
Target engagement dynamics can influence drugs' pharmacological effects. Kinetic parameters for drug:target interactions are often quantified by evaluating competition association experiments-measuring simultaneous protein binding of labelled tracers and unlabelled test compounds over time-with Motulsky-Mahan's "kinetics of competitive binding" model. Despite recent technical improvements, the current assay formats impose practical limitations to this approach. This study aims at the characterisation, understanding and prevention of these experimental constraints, and associated analytical challenges. Monte Carlo simulations were used to run virtual kinetic and equilibrium tracer binding and competition experiments in both normal and perturbed assay conditions. Data were fitted to standard equations derived from the mass action law (including Motulsky-Mahan's) and to extended versions aiming to cope with frequently observed deviations of the canonical traces. Results were compared to assess the precision and accuracy of these models and identify experimental factors influencing their performance. Key factors influencing the precision and accuracy of the Motulsky-Mahan model are the interplay between compound dissociation rates, measurement time and interval frequency, tracer concentration and binding kinetics and the relative abundance of equilibrium complexes in vehicle controls. Experimental results produced recommendations for better design of tracer characterisation experiments and new strategies to deal with systematic signal decay. Our data advances our comprehension of the Motulsky-Mahan kinetics of competitive binding models and provides experimental design recommendations, data analysis tools, and general guidelines for its practical application to in vitro pharmacology and drug screening.


Departmental and laboratory computing in two hospitals.
This report describes the departmental and laboratory use of integrated, hospital-wide computing systems at Beth Israel and Brigham and Women's hospitals in Boston. The systems have an important role in the admitting, outpatient, and medical records departments; in the clinical departments (blood bank, cardiology, neurophysiology, pathology, radiology, and pharmacy); in the clinical laboratories; and at Brigham and Women's Hospital, in the financial departments. Information that is collected in the computers from these departments and laboratories is available for viewing by clinicians at terminals located throughout each hospital and is used in compiling charges for each patient's account. The programs are heavily used. During a one-week study period, 742 departmental and laboratory workers at Beth Israel Hospital filed or edited information in patients' computerized records 137,526 times. During the same week, 984 departmental and laboratory workers at Brigham and Women's Hospital filed or edited information 293,367 times. After the computing systems were introduced, the time required to collect unpaid bills decreased substantially at both hospitals.


Evolutionary conserved Tyr169 stabilizes the β2-α2 loop of the prion protein.
Experimental evidence indicates that the primary structure of the β2-α2 loop region (residues 165-175) in mammalian prion proteins (PrP) influences the conversion from the cellular species (PrP(C)) to the β-sheet-rich aggregate. Here, we captured the transition of the β2-α2 loop from 310-helical turn to β turn by unbiased molecular dynamics simulations of the single-point mutant Y169G. Multiple conformations along the spontaneous transition of the mutant were then used as starting point for sampling of the free-energy surface of the wild type and other single-point mutants. Using two different methods for the determination of free energy profiles, we found that the barrier for the 310-helical turn to β turn transition of the wild type is higher by about 2.5 kcal/mol than for the Y169G mutant, which is due to favorable stacking of the aromatic rings of Y169 and F175, and a stable hydrogen bond between the side chains of Y169 and D178. The transition of the β2-α2 loop to β turn increases the solvent-exposure of the hydrophobic stretch 169-YSNQNNF-175. The simulations indicate that the strictly conserved Y169 in mammalian prion proteins stabilizes the 310-helical turn in the β2-α2 loop, thus hindering the conversion to an aggregation-prone conformation.


Arthroscopic rotator cuff repair with and without arthroscopic subacromial decompression: a prospective, randomized study of one-year outcomes.
We performed a prospective, randomized study to determine whether arthroscopic subacromial decompression changes the outcome of rotator cuff repair. We performed a power analysis to ensure statistical validity. Patients scheduled for arthroscopic rotator cuff repair were randomized to cuff repair with arthroscopic subacromial decompression (group 1) or without it (group 2). All other aspects of the surgical and postsurgical treatment were identical. We included patients with full-thickness tears limited to the supraspinatus tendon and a type 2 acromion. We excluded patients with prior surgery, those with larger tears involving two or more tendons, those with a type 1 or 3 acromion, those with workers' compensation claims, and those who had concomitant procedures (labral repair, acromioclavicular joint resection) There were 47 patients in group 1 and 46 in group 2. Minimum follow-up was 1 year (mean, 15.6 +/- 3.3 months). We recorded the American Shoulder and Elbow Surgeons (ASES) shoulder scores preoperatively and postoperatively. There was no statistical difference in postoperative ASES scores between group 1 (91.5 +/- 10.3) and group 2 (89.2 +/- 15.1) (P =.392). The change in ASES score over time did not differ between the two groups (61.1 vs 60.2, P =.363). In conclusion, within the parameters described above, arthroscopic subacromial decompression does not appear to change the functional outcome after arthroscopic repair of the rotator cuff.


Intramolecular symmetry-adapted perturbation theory with a single-determinant wavefunction.
We introduce an intramolecular energy decomposition scheme for analyzing non-covalent interactions within molecules in the spirit of symmetry-adapted perturbation theory (SAPT). The proposed intra-SAPT approach is based upon the Chemical Hamiltonian of Mayer [Int. J. Quantum Chem. 23(2), 341-363 (1983)] and the recently introduced zeroth-order wavefunction [J. F. Gonthier and C. Corminboeuf, J. Chem. Phys. 140(15), 154107 (2014)]. The scheme decomposes the interaction energy between weakly bound fragments located within the same molecule into physically meaningful components, i.e., electrostatic-exchange, induction, and dispersion. Here, we discuss the key steps of the approach and demonstrate that a single-determinant wavefunction can already deliver a detailed and insightful description of a wide range of intramolecular non-covalent phenomena such as hydrogen bonds, dihydrogen contacts, and π - π stacking interactions. Intra-SAPT is also used to shed the light on competing intra- and intermolecular interactions.


Sliced graphene foam films for dual-functional wearable strain sensors and switches.
The demand for wearable sensors is growing in many emerging fields, such as health monitoring, human-machine interfaces, robotics and personalized medicine. Here, we report the integration of skin-mountable, flexible, stretchable, dual-functional sensors and switches together with silicon-based electronics to create a novel healthcare system. We employ a facile approach to design highly stretchable graphene foam (GF)/PDMS composite films with tunable sensitivities and switching capabilities by simply controlling the thickness of GF. The GF/PDMS composite films deliver a high gauge factor of 24 at a 10% strain, tunable stretchability up to 70% and an excellent on/off switching ratio on the order of 1000. The highly reversible switching capability of the composite films is realized by identifying abnormal resistance changes at strains beyond a threshold value. To bridge the gap between signal transmission, wireless communication and post-processing in wearable devices, the sensors are combined with electronics, allowing data transmission to a smartphone using a custom-developed application consisting of a user-friendly interface. The novel approaches reported here offer a wide range of practical applications, including medical diagnosis, health monitoring and patient healthcare.


Effectiveness of home-based rehabilitation program in minimizing disability and secondary falls after a hip fracture: Protocol for a randomized controlled trial.
Hip fractures are a major health problem globally and are associated with increased morbidity, mortality, and substantial economic costs. Successful operative treatment of hip fracture patients is necessary for the optimization of post-op mobility and functional recovery of the patient. Rehabilitation after surgical stabilization of a hip fracture is crucial in order to restore pre-fracture function and to avoid long-term institutionalization. In particular ongoing exercise which targets balance can prevent up to 40% of falls. Therefore, we have designed a post-discharge home-based physical rehabilitation intervention program to minimize disability and falls in this high-risk elderly population. The study will be an open label, simple randomized controlled trial at a single hospital. The two arms will be equally allocated on a 1:1 ratio into intervention and control groups. The control arm will receive the usual standard postoperative rehabilitation. The intervention group will receive an extended home-based rehabilitation program twice a week continued for 3 months (12 weeks) after discharge. The Primary outcome of the study is occurrence of falls. Falls will be measured at 3, 6, 12, and 24 months by research-assistant follow-up telephone calls for both the groups. Mobility-related disability will be measured with a self-reported test at every routine follow-up for up to two years using a performance-based short battery tool. Negative binomial regression model will be used to compare number of falls in both the groups by computing incidence ratio rates. Approval for the conduction of this study has been taken from the Ethical Review Committee (ERC) of the institution. Evidences which will be obtained from this study will facilitate to propose changes in existing guidelines and policies for treating fall and hip fracture patients.Trial registrationThis trial is registered on clinicaltrials.gov ID: NCT04108793.


Antigen-specific expansion of human regulatory T cells as a major tolerance mechanism against mucosal fungi.
Foxp3(+) regulatory T cells (Treg) have a central role for keeping the balance between pro- and anti-inflammatory immune responses against chronically encountered antigens at mucosal sites. However, their antigen specificity especially in humans is largely unknown. Here we used a sensitive enrichment technology for antigen-reactive T cells to directly compare the conventional vs. regulatory CD4(+) T-cell response directed against two ubiquitous mucosal fungi, Aspergillus fumigatus and Candida albicans. In healthy humans, fungus-specific CD4(+)CD25(+)CD127(-)Foxp3(+) Treg are strongly expanded in peripheral blood and possess phenotypic, epigenetic and functional features of thymus-derived Treg. Intriguingly, for A. fumigatus, the strong Treg response contrasts with minimal conventional T-cell memory, indicating selective Treg expansion as an effective mechanism to prevent inappropriate immune activation in healthy individuals. By contrast, in subjects with A. fumigatus allergies, specific Th2 cells were strongly expanded despite the presence of specific Treg. Taken together, we demonstrate a largely expanded Treg population specific for mucosal fungi as part of the physiological human T-cell repertoire and identify a unique capacity of A. fumigatus to selectively generate Treg responses as a potentially important mechanism for the prevention of allergic reactions.


A method to fabricate disconnected silver nanostructures in 3D.
The standard nanofabrication toolkit includes techniques primarily aimed at creating 2D patterns in dielectric media. Creating metal patterns on a submicron scale requires a combination of nanofabrication tools and several material processing steps. For example, steps to create planar metal structures using ultraviolet photolithography and electron-beam lithography can include sample exposure, sample development, metal deposition, and metal liftoff. To create 3D metal structures, the sequence is repeated multiple times. The complexity and difficulty of stacking and aligning multiple layers limits practical implementations of 3D metal structuring using standard nanofabrication tools. Femtosecond-laser direct-writing has emerged as a pre-eminent technique for 3D nanofabrication.(1,2) Femtosecond lasers are frequently used to create 3D patterns in polymers and glasses.(3-7) However, 3D metal direct-writing remains a challenge. Here, we describe a method to fabricate silver nanostructures embedded inside a polymer matrix using a femtosecond laser centered at 800 nm. The method enables the fabrication of patterns not feasible using other techniques, such as 3D arrays of disconnected silver voxels.(8) Disconnected 3D metal patterns are useful for metamaterials where unit cells are not in contact with each other,(9) such as coupled metal dot(10,11)or coupled metal rod(12,13) resonators. Potential applications include negative index metamaterials, invisibility cloaks, and perfect lenses. In femtosecond-laser direct-writing, the laser wavelength is chosen such that photons are not linearly absorbed in the target medium. When the laser pulse duration is compressed to the femtosecond time scale and the radiation is tightly focused inside the target, the extremely high intensity induces nonlinear absorption. Multiple photons are absorbed simultaneously to cause electronic transitions that lead to material modification within the focused region. Using this approach, one can form structures in the bulk of a material rather than on its surface. Most work on 3D direct metal writing has focused on creating self-supported metal structures.(14-16) The method described here yields sub-micrometer silver structures that do not need to be self-supported because they are embedded inside a matrix. A doped polymer matrix is prepared using a mixture of silver nitrate (AgNO3), polyvinylpyrrolidone (PVP) and water (H2O). Samples are then patterned by irradiation with an 11-MHz femtosecond laser producing 50-fs pulses. During irradiation, photoreduction of silver ions is induced through nonlinear absorption, creating an aggregate of silver nanoparticles in the focal region. Using this approach we create silver patterns embedded in a doped PVP matrix. Adding 3D translation of the sample extends the patterning to three dimensions.


Monitoring pest insect traps by means of low-power image sensor technologies.
Monitoring pest insect populations is currently a key issue in agriculture and forestry protection. At the farm level, human operators typically must perform periodical surveys of the traps disseminated through the field. This is a labor-, time- and cost-consuming activity, in particular for large plantations or large forestry areas, so it would be of great advantage to have an affordable system capable of doing this task automatically in an accurate and a more efficient way. This paper proposes an autonomous monitoring system based on a low-cost image sensor that it is able to capture and send images of the trap contents to a remote control station with the periodicity demanded by the trapping application. Our autonomous monitoring system will be able to cover large areas with very low energy consumption. This issue would be the main key point in our study; since the operational live of the overall monitoring system should be extended to months of continuous operation without any kind of maintenance (i.e., battery replacement). The images delivered by image sensors would be time-stamped and processed in the control station to get the number of individuals found at each trap. All the information would be conveniently stored at the control station, and accessible via Internet by means of available network services at control station (WiFi, WiMax, 3G/4G, etc.).


Analysis of the Salmonella regulatory network suggests involvement of SsrB and H-NS in σ(E)-regulated SPI-2 gene expression.
The extracytoplasmic functioning sigma factor σ(E) is known to play an essential role for Salmonella enterica serovar Typhimurium to survive and proliferate in macrophages and mice. However, its regulatory network is not well-characterized, especially during infection. Here we used microarray to identify genes regulated by σ(E) in Salmonella grown in three conditions: a nutrient-rich condition and two others that mimic early and late intracellular infection. We found that in each condition σ(E) regulated different sets of genes, and notably, several global regulators. When comparing nutrient-rich and infection-like conditions, large changes were observed in the expression of genes involved in Salmonella pathogenesis island (SPI)-1 type-three secretion system (TTSS), SPI-2 TTSS, protein synthesis, and stress responses. In total, the expression of 58% of Salmonella genes was affected by σ(E) in at least one of the three conditions. An important finding is that σ(E) up-regulates SPI-2 genes, which are essential for Salmonella intracellular survival, by up-regulating SPI-2 activator ssrB expression at the early stage of infection and down-regulating SPI-2 repressor hns expression at a later stage. Moreover, σ(E) is capable of countering the silencing of H-NS, releasing the expression of SPI-2 genes. This connection between σ(E) and SPI-2 genes, combined with the global regulatory effect of σ(E), may account for the lethality of rpoE-deficient Salmonella in murine infection.


Estimating the causal influence of body mass index on risk of Parkinson disease: A Mendelian randomisation study.
Both positive and negative associations between higher body mass index (BMI) and Parkinson disease (PD) have been reported in observational studies, but it has been difficult to establish causality because of the possibility of residual confounding or reverse causation. To our knowledge, Mendelian randomisation (MR)-the use of genetic instrumental variables (IVs) to explore causal effects-has not previously been used to test the effect of BMI on PD. Two-sample MR was undertaken using genome-wide association (GWA) study data. The associations between the genetic instruments and BMI were obtained from the GIANT consortium and consisted of the per-allele difference in mean BMI for 77 independent variants that reached genome-wide significance. The per-allele difference in log-odds of PD for each of these variants was estimated from a recent meta-analysis, which included 13,708 cases of PD and 95,282 controls. The inverse-variance weighted method was used to estimate a pooled odds ratio (OR) for the effect of a 5-kg/m2 higher BMI on PD. Evidence of directional pleiotropy averaged across all variants was sought using MR-Egger regression. Frailty simulations were used to assess whether causal associations were affected by mortality selection. A combined genetic IV expected to confer a lifetime exposure of 5-kg/m2 higher BMI was associated with a lower risk of PD (OR 0.82, 95% CI 0.69-0.98). MR-Egger regression gave similar results, suggesting that directional pleiotropy was unlikely to be biasing the result (intercept 0.002; p = 0.654). However, the apparent protective influence of higher BMI could be at least partially induced by survival bias in the PD GWA study, as demonstrated by frailty simulations. Other important limitations of this application of MR include the inability to analyse non-linear associations, to undertake subgroup analyses, and to gain mechanistic insights. In this large study using two-sample MR, we found that variants known to influence BMI had effects on PD in a manner consistent with higher BMI leading to lower risk of PD. The mechanism underlying this apparent protective effect warrants further study.


The impact of an ambulance vehicle preparation service on the presence of bacteria: a service evaluation.
Around 300,000 patients a year in England acquire a healthcare-associated infection (HAI) while being cared for by the NHS. The contribution from NHS Ambulance Services is not known, but previous studies have identified the presence of pathogenic bacteria such as Methicillin-resistant Staphylococcus aureus (MRSA) and Enterococcus, including resistant strains in some cases, inside ambulances. To improve ambulance cleanliness, Yorkshire Ambulance Service NHS Trust (YAS) piloted an Ambulance Vehicle Preparation Service (AVPS) at two ambulance stations, where staff were tasked with ensuring every ambulance at these stations was cleaned every 24 hours. Adenosine triphosphate (ATP) bioluminescence testing was conducted on 16 ambulances at the two pilot AVPS stations and on 18 ambulances at four 'business as usual' (BAU) ambulance stations using a Hygiena SystemSURE luminometer. Swabs were obtained from 10 pre-selected locations inside each ambulance. Between November 2016 and August 2018, a total of 690 swabs were obtained and recorded from 34 ambulances. Overall, median relative light unit (RLU) values for both groups were < 100, with only the BAU group having an upper quartile value > 100. However, when stratified by swabbing area, three areas had a median RLU of > 100 in the BAU group: suction unit handle, steering wheel and airway seat shelf. In addition, the upper quartile RLU values for the grab rail above the stretcher and the passenger seat in the BAU group were also > 100. No swab areas had a median RLU > 100 in the AVPS group. A dedicated AVPS results in better cleaning of ambulance vehicles than the existing cleaning system utilising operational crews. The areas most likely to be contaminated are the suction unit handle, steering wheel, airway seat shelf and grab rails. The position of equipment and the materials that equipment are constructed from should have infection prevention and control (IPC) as a consideration.


Superficially invasive cervical squamous cell carcinoma metastatic to ovarian endometriotic cyst wall, a case report and brief review of the literature.
Although cases of cervical squamous cell carcinoma metastatic to the ovary have been previously documented, we report the first case of superficially invasive squamous cell carcinoma metastatic to the ovary. A 45-year-old woman with a two-year history of ovarian endometriosis confirmed by ultrasound underwent oophorectomy. On microscopic examination, a focus of malignant stratified epithelium, initially interpreted as transitional cell carcinoma, was identified within the endometriotic cyst wall. Examination of the hysterectomy specimen revealed superficially invasive squamous carcinoma of the cervix. In addition, two triploid, CD45-negative cells were detected during the analysis of the peripheral blood for circulating tumor cells (CTC). High-risk HPV was detected on the sections of endometriosis containing cancerous area by using hybrid capture 2 assay, supporting the diagnosis of metastatic squamous cell carcinoma originating from the uterine cervix. This is the first report of superficially invasive squamous cell carcinoma metastatic to the ovary. Such finding could be misdiagnosed as primary ovarian transitional cell carcinoma, squamous cell carcinoma originating from metaplastic epithelium within endometriosis, or squamous cell carcinoma arising in a teratoma.


The value of inflammatory markers to diagnose and monitor diabetic foot osteomyelitis.
In this study, we assessed the effectiveness of inflammatory markers to diagnose and monitor the treatment of osteomyelitis in the diabetic foot. We evaluated 35 consecutive patients admitted to our hospital with infected foot ulcers. Patients were divided in two groups based on the results of bone culture and histopathology: osteomyelitis and no osteomyelitis. The erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), procalcitonin (PCT), interleukin-6 (IL-6), interleukin-8 (IL-8), tumor necrosis factor alpha (TNFα), monocyte chemotactic protein-1 (MCP-1) and macrophage inflammatory protein-1 alpha (MIP1α) were measured at baseline after 3 and 6 weeks of standard therapy. PCT levels in the osteomyelitis group were significantly higher at baseline than in the group with no osteomyelitis (P = 0·049). There were no significant differences between the two groups in the levels of the other markers. CRP, ESR, PCT and IL-6 levels significantly declined in the group with osteomyelitis after starting therapy, while MCP-1 increased (P = 0·002). TNFα and MIP1α levels were below range in 80 out of 97 samples and therefore not reported. Our results suggest that PCT might be useful to distinguish osteomyelitis in infected foot ulcers. CRP, ESR, PCT and IL-6 are valuable when monitoring the effect of therapy.


Corrections and clarifications.
In the news article "FDA puts the brakes on xenotransplants" by Rachel Nowak (5 May, p. 630) the description of the response of Columbia Presbyterian Medical Center in New York City to one of its investigators' proposal to test baboon-to-human heart transplants was incorrect. In fact, Columbia convened one panel of outside experts to advise its internal committees on the risks of xenotransplantation, and has since retained two outside experts as consultants to Robert Michler, the principle investigator for the xenotransplant trials. The institution also recommended that a group of experts on emerging infections be convened to examine the risk of viral transmission associated with xenotransplantation (Letters, 21 Apr., p. 349).


Choline kinase beta is required for normal endochondral bone formation.
Choline kinase has three isoforms encoded by the genes Chka and Chkb. Inactivation of Chka in mice results in embryonic lethality, whereas Chkb(-/-) mice display neonatal forelimb bone deformations. To understand the mechanisms underlying the bone deformations, we compared the biology and biochemistry of bone formation from embryonic to young adult wild-type (WT) and Chkb(-/-) mice. The deformations are specific to the radius and ulna during the late embryonic stage. The radius and ulna of Chkb(-/-) mice display expanded hypertrophic zones, unorganized proliferative columns in their growth plates, and delayed formation of primary ossification centers. The differentiation of chondrocytes of Chkb(-/-) mice was impaired, as was chondrocyte proliferation and expression of matrix metalloproteinases 9 and 13. In chondrocytes from Chkb(-/-) mice, phosphatidylcholine was slightly lower than in WT mice whereas the amount of phosphocholine was decreased by approximately 75%. In addition, the radius and ulna from Chkb(-/-) mice contained fewer osteoclasts along the cartilage/bone interface. Chkb has a critical role in the normal embryogenic formation of the radius and ulna in mice. Our data indicate that choline kinase beta plays an important role in endochondral bone formation by modulating growth plate physiology.


Existing creatinine-based equations overestimate glomerular filtration rate in Indians.
Accurate estimation of glomerular filtration rate (GFR) is important for diagnosis and risk stratification in chronic kidney disease and for selection of living donors. Ethnic differences have required correction factors in the originally developed creatinine-based GFR estimation equations for populations around the world. Existing equations have not been validated in the vegetarian Indian population. We examined the performance of creatinine and cystatin-based GFR estimating equations in Indians. GFR was measured by urinary clearance of inulin. Serum creatinine was measured using IDMS-traceable Jaffe's and enzymatic assays, and cystatin C by colloidal gold immunoassay. Dietary protein intake was calculated by measuring urinary nitrogen appearance. Bias, precision and accuracy were calculated for the eGFR equations. A total of 130 participants (63 healthy kidney donors and 67 with CKD) were studied. About 50% were vegetarians, and the remainder ate meat 3.8 times every month. The average creatinine excretion were 14.7 mg/kg/day (95% CI: 13.5 to 15.9 mg/kg/day) and 12.4 mg/kg/day (95% CI: 11.2 to 13.6 mg/kg/day) in males and females, respectively. The average daily protein intake was 46.1 g/day (95% CI: 43.2 to 48.8 g/day). The mean mGFR in the study population was 51.66 ± 31.68 ml/min/1.73m2. All creatinine-based eGFR equations overestimated GFR (p < 0.01 for each creatinine based eGFR equation). However, eGFR by CKD-EPICys was not significantly different from mGFR (p = 0.38). The CKD-EPICys exhibited lowest bias [mean bias: -3.53 ± 14.70 ml/min/1.73m2 (95% CI: -0.608 to -0.98)] and highest accuracy (P30: 74.6%). The GFR in the healthy population was 79.44 ± 20.19 (range: 41.90-134.50) ml/min/1.73m2. Existing creatinine-based GFR estimating equations overestimate GFR in Indians. An appropriately powered study is needed to develop either a correction factor or a new equation for accurate assessment of kidney function in the Indian population.


Coupling to polymeric scaffolds stabilizes biofunctional peptides for intracellular applications.
Here, we demonstrate that coupling to N-hydroxypropyl methacrylamide (HPMA) copolymer greatly enhances the activity of apoptosis-inducing peptides inside cells. Peptides corresponding to the BH3 domain of Bid were coupled to a thioester-activated HPMA (28.5 kDa) via native chemical ligation in a simple one-pot synthesis. Peptides and polymer conjugates were introduced into cells either by electroporation or by conjugation to the cell-penetrating peptide nona-arginine. The molecular basis of the increased activity is elucidated in detail. Loading efficiency and intracellular residence time were assessed by confocal microscopy. Fluorescence correlation spectroscopy was used as a separation-free analytical technique to determine proteolytic degradation in crude cell lysates. HPMA conjugation strongly increased the half-life of the peptides in crude cell lysates and inside cells, revealing proteolytic protection as the basis for higher activity.


Potential exposure to Australian bat lyssavirus, Queensland, 1996-1999.
Two human deaths caused by Australian bat lyssavirus (ABL) infection have been reported since 1996. Information was obtained from 205 persons (mostly adults from south Brisbane and the South Coast of Queensland), who reported potential ABL exposure to the Brisbane Southside Public Health Unit from November 1,1996, to January 31, 1999. Volunteer animal handlers accounted for 39% of potential exposures, their family members for 12%, professional animal handlers for 14%, community members who intentionally handled bats for 31%, and community members with contacts initiated by bats for 4%. The prevalence of Lyssavirus detected by fluorescent antibody test in 366 sick, injured, or orphaned bats from the area was 6%. Sequelae of exposure, including the requirement for expensive postexposure prophylaxis, may be reduced by educating bat handlers and the public of the risks involved in handling Australian bats.


The Clinica del Lavoro of Milan provided several contributions to industrial hygiene and occupational toxicology during the twentieth century. Describe the first years of the laboratory of industrial hygiene of Milan through three figures who played a leading role: Enrico Carlo Vigliani, Nicola Zurlo and Gianmario Cavagna. Scientific literature of the period 1948-1970 was investigated, also interviewing first-hand witnesses of that period. Enrico Vigliani was the first European scholar to understand the importance of a laboratory of industrial hygiene within his institution. Thanks to the support of private (Montecatini) and public (INAIL) institutions he succeeded in creating a laboratory in 1948. Nicola Zurlo, who directed this structure in the first thirty years, conducted innovative studies on chronic mercury intoxication, lead intoxication and silicosis, designing and creating instruments for capturing and analyzing atmospheric dust and protection devices. He conducted analysis of the health effects of organophosphorus insecticides and started to study the air pollution. Zurlo also provided an epistemological and methodological content to the discipline. Gianmario Cavagna, one of the first Italian toxicologists, contributed to the discovery of the origin of fevers caused by the inhalation of metal fumes and to the studies on the pathogenesis of byssinosis, hypothesizing a role of bacterial endotoxins in the genesis of this disease. The contributions provided by these three protagonists to industrial hygiene and occupational toxicology were relevant and made in those years the Clinica del Lavoro of Milan as a landmark, not only in Italy but also abroad.


Determination of the nano-scaled contact area of staphylococcal cells.
Bacterial adhesion is a crucial step during the development of infections as well as the formation of biofilms. Hence, fundamental research of bacterial adhesion mechanisms is of utmost importance. So far, less is known about the size of the contact area between bacterial cells and a surface. This gap will be filled by this study using a single-cell force spectroscopy-based method to investigate the contact area between a single bacterial cell of Staphylococcus aureus and a solid substrate. The technique relies on the strong influence of the hydrophobic interaction on bacterial adhesion: by incrementally crossing a very sharp hydrophobic/hydrophilic interface while performing force-distance curves with a single bacterial probe, the bacterial contact area can be determined. Assuming circular contact areas, their radii - determined in our experiments - are in the range from tens of nanometers to a few hundred nanometers. The contact area can be slightly enlarged by a larger load force, yet does not resemble a Hertzian contact, rather, the enlargement is a property of the individual bacterial cell. Additionally, Staphylococcus carnosus has been probed, which is less adherent than S. aureus, yet both bacteria exhibit a similar contact area size. This corroborates the notion that the adhesive strength of bacteria is not a matter of contact area, but rather a matter of which and how many molecules of the bacterial species' cell wall form the contact. Moreover, our method of determining the contact area can be applied to other microorganisms and the results might also be useful for studies using nanoparticles covered with soft, macromolecular coatings.


Blood-Stage Parasitaemia and Age Determine Plasmodium falciparum and P. vivax Gametocytaemia in Papua New Guinea.
A better understanding of human-to-mosquito transmission is crucial to control malaria. In order to assess factors associated with gametocyte carriage, 2083 samples were collected in a cross-sectional survey in Papua New Guinea. Plasmodium species were detected by light microscopy and qPCR and gametocytes by detection of pfs25 and pvs25 mRNA transcripts by reverse-transcriptase PCR (qRT-PCR). The parasite prevalence by PCR was 18.5% for Plasmodium falciparum and 13.0% for P. vivax. 52.5% of all infections were submicroscopic. Gametocytes were detected in 60% of P. falciparum-positive and 51% of P. vivax-positive samples. Each 10-fold increase in parasite density led to a 1.8-fold and 3.3-fold increase in the odds of carrying P. falciparum and P. vivax gametocytes. Thus the proportion of gametocyte positive and gametocyte densities was highest in young children carrying high asexual parasite densities and in symptomatic individuals. Dilution series of gametocytes allowed absolute quantification of gametocyte densities by qRT-PCR and showed that pvs25 expression is 10-20 fold lower than pfs25 expression. Between 2006 and 2010 parasite prevalence in the study site has decreased by half. 90% of the remaining infections were asymptomatic and likely constitute an important reservoir of transmission. However, mean gametocyte densities were low (approx. 1-2 gametocyte/μL) and it remains to be determined to what extent low-density gametocyte positive individuals are infective to mosquitos.


Biotransformation of ferulic acid to 4-vinylguaiacol by Enterobacter soli and E. aerogenes.
We investigated the conversion of ferulic acid to 4-vinylguaiacol (4-VG), vanillin, vanillyl alcohol, and vanillic acid by five Enterobacter strains. These high-value chemicals are usually synthesized by chemical methods but biological synthesis adds market value. Ferulic acid, a relatively inexpensive component of agricultural crops, is plentiful in corn hulls, cereal bran, and sugar-beet pulp. Two Enterobacter strains, E. soli, and E. aerogenes, accumulated 550-600 ppm amounts of 4-VG when grown in media containing 1,000 ppm ferulic acid; no accumulations were observed with the other strains. Decreasing the amount of ferulic acid present in the media increased the conversion efficiency. When ferulic acid was supplied in 500, 250, or 125 ppm amounts E. aerogenes converted ~72 % of the ferulic acid present to 4-VG while E. soli converted ~100 % of the ferulic acid to 4-VG when supplied with 250 or 125 ppm amounts of ferulic acid. Also, lowering the pH improved the conversion efficiency. At pH 5.0 E. aerogenes converted ~84 % and E. soli converted ~100 % of 1,000 ppm ferulic acid to 4-VG. Only small, 1-5 ppm, accumulations of vanillin, vanillyl alcohol, and vanillic acid were observed. E. soli has a putative phenolic acid decarboxylase (PAD) that is 168 amino acids long and is similar to PADs in other enterobacteriales; this protein is likely involved in the bioconversion of ferulic acid to 4-VG. E. soli or E. aerogenes might be useful as a means of biotransforming ferulic acid to 4-VG.


High-dimensional analysis of evolutionary autonomous agents.
This article presents a new approach to the important challenge of localizing function in a neurocontroller. The approach is based on the basic functional contribution analysis (FCA) presented earlier, which assigns contribution values to the elements of the network, such that the ability to predict the network's performance in response to multi-unit lesions is maximized. These contribution values quantify the importance of each element to the tasks the agent performs. Here we present a generalization of the basic FCA to high-dimensional analysis, using high-order compound elements. Such elements are composed of conjunctions of simple elements. Their usage enables the explicit expression of sets of neurons or synapses whose contributions are interdependent, a prerequisite for localizing the function of complex neurocontrollers. High-dimensional FCA is shown to significantly improve on the accuracy of the basic analysis, to provide new insights concerning the main subsets of simple elements in the network that interact in a complex nonlinear manner, and to systematically reveal the types of interactions that characterize the evolved neurocontroller.


An autonomous wearable system for diurnal sweat biomarker data acquisition.
To track dynamically varying and physiologically relevant biomarker profiles in sweat, autonomous wearable platforms are required to periodically sample and analyze sweat with minimal or no user intervention. Previously reported sweat sensors are functionally limited to capturing biomarker information at one time-point/period, thereby necessitating repeated user intervention to increase the temporal granularity of biomarker data. Accordingly, we present a compact multi-compartment wearable system, where each compartment can be activated to autonomously induce/modulate sweat secretion (via iontophoretic actuation) and analyze sweat at set time points. This system was developed following a hybrid-flex design and a vertical integration scheme-integrating the required functional modules: miniaturized iontophoresis interfaces, adhesive thin film microfluidic-sensing module, and control/readout electronics. The system was deployed in a human subject study to track the diurnal variation of sweat glucose levels in relation to the daily food intake. The demonstrated autonomous operation for diurnal sweat biomarker data acquisition illustrates the system's suitability for large-scale and longitudinal personal health monitoring applications.


Severity of Giardia infection associated with post-infectious fatigue and abdominal symptoms two years after.
A high rate of post-infectious fatigue and abdominal symptoms two years after a waterborne outbreak of giardiasis in Bergen, Norway in 2004 has previously been reported. The aim of this report was to identify risk factors associated with such manifestations. All laboratory confirmed cases of giardiasis (n = 1262) during the outbreak in Bergen in 2004 received a postal questionnaire two years after. Degree of post-infectious abdominal symptoms and fatigue, as well as previous abdominal problems, was recorded. In the statistical analyses number of treatment courses, treatment refractory infection, delayed education and sick leave were used as indices of protracted and severe Giardia infection. Age, gender, previous abdominal problems and symptoms during infection were also analysed as possible risk factors. Simple and multiple ordinal logistic regression models were used for the analyses. The response rate was 81% (1017/1262), 64% were women and median age was 31 years (range 3-93), compared to 61% women and 30 years (range 2-93) among all 1262 cases. Factors in multiple regression analysis significantly associated with abdominal symptoms two years after infection were: More than one treatment course, treatment refractory infection, delayed education, bloating and female gender. Abdominal problems prior to Giardia infection were not associated with post-infectious abdominal symptoms. More than one treatment course, delayed education, sick leave more than 2 weeks, and malaise at the time of infection, were significantly associated with fatigue in the multiple regression analysis, as were increasing age and previous abdominal problems. Protracted and severe giardiasis seemed to be a risk factor for post-infectious fatigue and abdominal symptoms two years after clearing the Giardia infection.


Mesenteric Ischemia: Concepts of Care for the Bedside Nurse.
Bedside nurses require excellent observational and assessment skills. Being knowledgeable about common diseases that often affect the population is essential; however, it is often more challenging to be informed about life-threatening conditions that rarely develop. Such is the case with mesenteric ischemia, a rare but complicated condition, which is difficult to diagnose, carries a degree of mortality of about 40%-70%, and is more common in people older than 50 years. Mesenteric ischemia requires that nurses be aware of the different etiologies and presentations of this uncommon disease to alert physicians of changes in a patient's condition and detect clues that may assist with an accurate and prompt diagnosis. Being educated about the diagnostic procedures, possible outcomes, and anticipated care and needs of patients affected by this medical complication is imperative to maintain the health status and well-being of these patients and their families.


A 6-month observational study of changes in objectively measured physical activity during weight loss in dogs.
To evaluate long-term changes in physical activity and sedentary behaviour during weight loss in dogs. Sixteen overweight and obese dogs undergoing a 6-month calorie-controlled weight-loss programme wore Actigraph GT3X accelerometers (Actigraph, FL) for three consecutive days each month for the duration of the programme. Total volume of physical activity and time spent in sedentary behaviour, light-moderate intensity physical activity and vigorous intensity physical activity were extracted from the accelerometer data and compared from baseline to month 6. Valid accelerometry data were returned for 14 of 16 dogs. Mean percentage weight loss over 6 months was 15% of initial bodyweight. There was no marked increase in any of the physical activity outcomes or reduction in sedentary behaviour. Substantial weight loss was not associated with a spontaneous increase in physical activity or reduction in sedentary behaviour. Although the dogs in this study lost a substantial amount of bodyweight without a measured increase in physical activity, dog owners should still be encouraged to facilitate increased physical activity in their dogs owing to the wide range of benefits associated with physical activity other than weight loss.


Cross-regulation of viral kinases with cyclin A secures shutoff of host DNA synthesis.
Herpesviruses encode conserved protein kinases (CHPKs) to stimulate phosphorylation-sensitive processes during infection. How CHPKs bind to cellular factors and how this impacts their regulatory functions is poorly understood. Here, we use quantitative proteomics to determine cellular interaction partners of human herpesvirus (HHV) CHPKs. We find that CHPKs can target key regulators of transcription and replication. The interaction with Cyclin A and associated factors is identified as a signature of β-herpesvirus kinases. Cyclin A is recruited via RXL motifs that overlap with nuclear localization signals (NLS) in the non-catalytic N termini. This architecture is conserved in HHV6, HHV7 and rodent cytomegaloviruses. Cyclin A binding competes with NLS function, enabling dynamic changes in CHPK localization and substrate phosphorylation. The cytomegalovirus kinase M97 sequesters Cyclin A in the cytosol, which is essential for viral inhibition of cellular replication. Our data highlight a fine-tuned and physiologically important interplay between a cellular cyclin and viral kinases.


External regulation of controlled polymerizations.
Polymer chemists, through advances in controlled polymerization techniques and reliable post-functionalization methods, now have the tools to create materials of almost infinite variety and architecture. Many relevant challenges in materials science, however, require not only functional polymers but also on-demand access to the properties and performance they provide. The power of such temporal and spatial control of polymerization can be found in nature, where the production of proteins, nucleic acids, and polysaccharides helps regulate multicomponent systems and maintain homeostasis. Here we review existing strategies for temporal control of polymerizations through external stimuli including chemical reagents, applied voltage, light, and mechanical force. Recent work illustrates the considerable potential for this emerging field and provides a coherent vision and set of criteria for pursuing future strategies for regulating controlled polymerizations.


Comparative neurohistological observations on the intrinsic innervation of pancreatic artery (blood vessel) in the representative of bird and mammal.
The present investigation was undertaken for the study of the comparative neurohistological observations of pancreatic-Artery (blood vessel) in Gallus domesticus (white leghorn) and Suncus murinus (Indian musk shrew) by cholinesterase technique. In Gallus, the innervation of arteries (blood vessel) was by a good number of non-myelinated nerevs, which formed the periarterial plexus in participation with the postganglionic fibres at one end and the fibres of the nerve cells of the other end. In Suncus, the periarterial and AChE-positive ganglia were arranged in chain-like fashion on the periphery of the artery. Periarterial plexus was formed by tortuous, myelinated nerves and the nerve fibres of the ganglia.


Single-Protein Collapse Determines Phase Equilibria of a Biological Condensate.
Recent advances in microscopy of living cells have established membraneless organelles as critical elements of diverse biological processes. The body of experimental work suggests that formation of such organelles is driven by liquid-liquid phase separation, a physical process that has been studied extensively for both simple liquids and mixtures of polymers. Here, we combine molecular dynamics simulations with polymer theory to show that the thermodynamic behavior of one particular biomolecular condensate-fused in sarcoma (FUS)-can be quantitatively accounted for at the level of the chain collapse theory. First, we show that a particle-based molecular dynamics model can reproduce known phase separation properties of a FUS condensate, including its critical concentration and susceptibility to mutations. Next, we obtain a polymer physics representation of a FUS condensate by examining the behavior of a single FUS protein as a function of temperature. We use the chain collapse theory to determine the thermodynamic properties of the condensate and to characterize changes in the single-chain conformation at the onset of phase separation. Altogether, our findings suggest that the phase behavior of FUS condensates can be explained by the properties of individual FUS proteins and that the change in the FUS conformation is the main force driving for the phase separation.


Statistics of specular points at a randomly rough surface.
Statistical parameters are obtained for an ensemble of specular points at a randomly rough Gaussian statistically isotropic surface at normal incidence. The joint probability density functions (PDFs) of specular point heights and total curvatures are derived separately for maxima, minima, and saddle points. The joint PDFs of brightness and surface elevations of specular points of different types are obtained analytically in an explicit form.


Insights into reference point indentation involving human cortical bone: sensitivity to tissue anisotropy and mechanical behavior.
Reference point indentation (RPI) is a microindentation technique involving 20 cycles of loading in "force-control" that can directly assess a patient׳s bone tissue properties. Even though preliminary clinical studies indicate a capability for fracture discrimination, little is known about what mechanical behavior the various RPI properties characterize and how these properties relate to traditional mechanical properties of bone. To address this, the present study investigated the sensitivity of RPI properties to anatomical location and tissue organization as well as examined to what extent RPI measurements explain the intrinsic mechanical properties of human cortical bone. Multiple indents with a target force of 10N were done in 2 orthogonal directions (longitudinal and transverse) per quadrant (anterior, medial, posterior, and lateral) of the femoral mid-shaft acquired from 26 donors (25-101 years old). Additional RPI measurements were acquired for 3 orthogonal directions (medial only). Independent of age, most RPI properties did not vary among these locations, but they did exhibit transverse isotropy such that resistance to indentation is greater in the longitudinal (axial) direction than in the transverse direction (radial or circumferential). Next, beam specimens (~2mm×5mm×40mm) were extracted from the medial cortex of femoral mid-shafts, acquired from 34 donors (21-99 years old). After monotonically loading the specimens in three-point bending to failure, RPI properties were acquired from an adjacent region outside the span. Indent direction was orthogonal to the bending axis. A significant inverse relationship was found between resistance to indentation and the apparent-level mechanical properties. Indentation distance increase (IDI) and a linear combination of IDI and the loading slope, averaged over cycles 3 through 20, provided the best explanation of the variance in ultimate stress (r(2)=0.25, p=0.003) and toughness (r(2)=0.35, p=0.004), respectively. With a transverse isotropic behavior akin to tissue hardness and modulus as determined by micro- and nano-indentation and a significant association with toughness, RPI properties are likely influenced by both elastic and plastic behavior of bone tissue.


Sexual Identity and Behavior Among U.S. High School Students, 2005-2015.
Sexual orientation is a multidimensional construct which is increasingly recognized as an important demographic characteristic in population health research. For this study, weighted Youth Risk Behavior Survey data were pooled across 47 jurisdictions biennially from 2005 to 2015, resulting in a national sample of 98 jurisdiction-years (344,815 students). Respondents were a median of 15.5 years, 49.9% male, and 48.8% White. Sexual identity and behavior trends from 2005 to 2015 were assessed with logistic regression analysis. Overall, 13.9% of females and 7.0% of males identified as lesbian, gay, bisexual (LGB), or not sure, while 9.1% of females and 4.2% of males indicated both same-and-different-sex behavior or same-sex behavior. In total, 17.0% of female and 8.5% of male youth reported non-heterosexual (LGB or not sure) sexual identity, same-sex sexual behavior, or both. LGB youth were approximately twice as likely as other youth to report lifetime sexual behavior. White and Asian youth were less likely to report non-heterosexual identity and/or have engaged in same-sex sexual behaviors than youth of other races/ethnicities. Prevalence of non-heterosexual identities increased over time for both sexes, but only female youth reported significantly more same-sex behavior over time. This is the first study to simultaneously assess adolescent sexual identity and behavior over time within a national dataset. These findings are critical for understanding the sexual health needs of adolescents and for informing sexual health policy and practice.


Neoadjuvant imatinib for borderline resectable GIST.
A 36-year-old woman presented to the emergency department with black stools and syncope. Her hemoglobin was 7.0 and her red blood cells were microcytic. Upper endoscopy did not identify a clear source of bleeding, but a bulge in the third portion of the duodenum was noted. A CT scan showed a large extraintestinal mass, and follow-up esophagogastroduodenoscopy/endoscopic ultrasound with biopsy revealed a spindle cell neoplasm, consistent with gastrointestinal stromal tumor (GIST). Because of the size of the lesion and association with the superior mesenteric vein and common bile duct, she was referred to medical oncology for consideration of neoadjuvant imatinib. Neoadjuvant tyrosine kinase inhibitor therapy for GISTs is emerging as a viable treatment strategy for borderline resectable tumors, although the dose, duration, and optimal imaging modalities have not been clearly established. Recent pathologic and radiographic data have provided insight into the mechanism and kinetics of this approach. This case report presents a patient for whom surgery was facilitated using neoadjuvant imatinib.


Sleep-disordered breathing in preschool children is associated with behavioral, but not cognitive, impairments.
Sleep-disordered breathing (SDB) has been associated with impaired cognitive and behavioral function in school children; however, there have been limited studies in preschool children when the incidence of the disorder peaks. Thus, the aim of this study was to compare cognitive and behavioral functions of preschool children with SDB to those of non-snoring control children. A clinical sample of 3-5 year-old children (primary snoring [PS], n=60; mild obstructive sleep apnea syndrome [OSAS], n=32; moderate/severe [MS] OSAS, n=24) and a community sample of non-snoring control children (n=37) were studied with overnight polysomnography. Cognitive performance and behavioral information were collected. Children with PS and mild OSAS had poorer behavior than controls on numerous measures (p<.05-p<.001), and on some measures they had poorer behavior than the MS OSAS group (p<.05). In contrast, all groups performed similarly on cognitive assessment. Outcomes related more to sleep than respiratory measures. SDB of any severity was associated with poorer behavior but not cognitive performance. The lack of significant cognitive impairment in this age group may have identified a "window of opportunity" where early treatment may prevent deficits arising later in childhood.


Liver function tests and fibrosis scores in a rural population in Africa: a cross-sectional study to estimate the burden of disease and associated risk factors.
Liver disease is a major cause of morbidity and mortality in sub-Saharan Africa, but its prevalence, distribution and aetiology have not been well characterised. We therefore set out to examine liver function tests (LFTs) and liver fibrosis scores in a rural African population. We undertook a cross-sectional survey of LFTs. We classified abnormal LFTs based on reference ranges set in America and in Africa. We derived fibrosis scores (aspartate aminotransferase (AST) to Platelet Ratio Index (APRI), fibrosis-4, gamma-glutamyl transferase (GGT) to platelet ratio (GPR), red cell distribution width to platelet ratio and S-index). We collected information about alcohol intake, and infection with HIV, hepatitis B virus (HBV) and hepatitis C virus (HCV). We studied a population cohort in South-Western Uganda. Data were available for 8099 adults (median age 30 years; 56% female). The prevalence of HBV, HCV and HIV infection was 3%, 0.2% and 8%, respectively. The prevalence of abnormal LFTs was higher based on the American reference range compared with the African reference range (eg, for AST 13% vs 3%, respectively). Elevated AST/ALT ratio was significantly associated with self-reported alcohol consumption (p<0.001), and the overall prevalence of AST/ALT ratio >2 was 11% (suggesting alcoholic hepatitis). The highest prevalence of fibrosis was predicted by the GPR score, with 24% of the population falling above the threshold for fibrosis. There was an association between the presence of HIV or HBV and raised GPR (p=0.005) and S-index (p<0.001). By multivariate analysis, elevated LFTs and fibrosis scores were most consistently associated with older age, male sex, being under-weight, HIV or HBV infection and alcohol consumption. Further work is required to determine normal reference ranges for LFTs in this setting, to evaluate the specificity and sensitivity of fibrosis scores and to determine the aetiology of liver disease.


Technical feasibility and life cycle assessment of an industrial waste as stabilizing product for unpaved roads, and influence of packaging.
The use of industrial solid wastes with a high content of SiO2 and Al2O3, called "precursors", is often studied in the construction industry when combined with NaOH as "activator". The precursor and activator system is generally proposed as a binder material with similar characteristics to Portland cement. In this work, we technically and environmentally evaluated such a system elaborated with an industrial waste: coal ash with caustic soda in solid state. This product, mixed with the soil, acts as a stabilizer to increase the capacity of load support, allowing the improvement of the conditions of performance in low volume traffic roads. An experimental design applied to the stabilizing product showed the incidence of different factors on the load carrying capacity response: packaging material, type of seal, baling moisture and storage humidity. The application of the stabilizer product was found to increase the resistance of the ground over a 500%. Finally, the environmental aspects were evaluated through a simplified Life Cycle Assessment methodology (LCA), the scope of the study was restricted to cradle to gate, collecting data up to the packaged stabilizing product. The results showed that the highest impacts were caused, for most impact categories, by NaOH production, and transport was relevant as well.


Capturing the multiscale dynamics of membrane protein complexes with all-atom, mixed-resolution, and coarse-grained models.
The structures and dynamics of protein complexes are often challenging to model in heterogeneous environments such as biological membranes. Herein, we meet this fundamental challenge at attainable cost with all-atom, mixed-resolution, and coarse-grained models of vital membrane proteins. We systematically simulated five complex models formed by two distinct G protein-coupled receptors (GPCRs) in the lipid-bilayer membrane on the ns-to-μs timescales. These models, which suggest the swinging motion of an intracellular loop, for the first time, provide the molecular details for the regulatory role of such a loop. For the models at different resolutions, we observed consistent structural stability but various levels of speed-ups in protein dynamics. The mixed-resolution and coarse-grained models show two and four times faster protein diffusion than the all-atom models, in addition to a 4- and 400-fold speed-up in the simulation performance. Furthermore, by elucidating the strengths and challenges of combining all-atom models with reduced resolution models, this study can serve as a guide to simulating other complex systems in heterogeneous environments efficiently.


The meaning of middle-aged female spouses' lived experience of the relationship with a partner who has suffered a stroke, during the first year postdischarge.
Stroke consequences present a great long-term challenge to the spouses of the stroke sufferer. A longitudinal study with a phenomenological hermeneutic approach was used to illuminate the meanings of middle-aged female spouses' lived experience of their relationship with a partner who has suffered a stroke, during the first year postdischarge. Four middle-aged female spouses of stroke sufferers participated in the study. Narrative interviews were conducted 1, 6 and 12 month postdischarge (total of 12 interviews), audio-taped and transcribed verbatim to a text and interpreted by a phenomenological hermeneutic method inspired by Ricoeur. The findings showed a process over time during which spouses come to know, recognize and acknowledge the residual changes in their partners' cognitive and emotional conditions and the impact on their relationship. Spouses showed grief due to the loss of the marital relationship they once had and anxiety that they would not be able to continue in an undesired relationship in the future. Even if the partner is still alive, there is a loss to grieve and to be understood, an important meaning of the transition process in the relationship during the first year after the partner's discharge.


Pediatric anesthesia and neurotoxicity: what the radiologist needs to know.
The use of cross-sectional imaging in the pediatric population continues to rise, particularly the use of MRI. Limiting motion artifact requires cooperative subjects who do not move during imaging, so there has been an increase in the need for pediatric sedation or anesthesia. Over the last decade, concern has increased that exposure to anesthesia might be associated with long-term cognitive deficits. In this review we report current understanding of the effects of anesthesia on the pediatric population, with special focus on long-term developmental and cognitive outcomes, and suggest how radiologists can use new technologies or imaging strategies to mitigate or minimize these potential risks.


Chronic subdural hematoma pathophysiology: a unifying theory for a dynamic process.
Chronic subdural hematoma pathophysiology has been extensively studied and discussed. In the last decades, optic and electron microscope observations have successfully described its histopathology and the ultrastructure of internal membranes. Moreover, recent biochemical studies have identified a number of important pathways involved in its development and evolution. Our aim was to review recent literature regarding histopathology, ultrastructure and biochemichal pathways and supply a unifying theory about chronic subdural hematoma pathophysiology. The starting point of chronic subdural hematoma is a mechanical injury. The evolution of the pathology is due to the exclusive anatomy of the dura-arachnoid interface. This is a mechanically weak layer. Fibroblasts contained in this region produce an inflammatory reaction with neoangiogenesis and fibrinolysis. Biochemical pathways involved in these reactions is complex and could contain a number of pharmacological targets. The hematoma evolves in different stages thus recent outlooks consider chronic subdural hematoma as a dynamic process. One of the key points for a good outcome and a low recurrence rate may be the timing of the surgical treatment in relation of hematoma natural history. Surgery performed during active inflammatory stages may be less effective in terms of clinical outcome and recurrence rate.


Knowledge sharing in the health scenario.
The understanding of certain data often requires the collection of similar data from different places to be analysed and interpreted. Interoperability standards and ontologies, are facilitating data interchange around the world. However, beyond the existing networks and advances for data transfer, data sharing protocols to support multilateral agreements are useful to exploit the knowledge of distributed Data Warehouses. The access to a certain data set in a federated Data Warehouse may be constrained by the requirement to deliver another specific data set. When bilateral agreements between two nodes of a network are not enough to solve the constraints for accessing to a certain data set, multilateral agreements for data exchange are needed. We present the implementation of a Multi-Agent System for multilateral exchange agreements of clinical data, and evaluate how those multilateral agreements increase the percentage of data collected by a single node from the total amount of data available in the network. Different strategies to reduce the number of messages needed to achieve an agreement are also considered. The results show that with this collaborative sharing scenario the percentage of data collected dramaticaly improve from bilateral agreements to multilateral ones, up to reach almost all data available in the network.


High-frequency C-type virus induction by inhibitors of protein synthesis.
When inhibitors of protein synthesis are added to BALB/c mouse cells in culture, induction of naturally integrated C-type RNA virus occurs in a high percentage of cells. The action of protein synthesis inhibitors differs from that of halogenated pyrimidines, another class of virus inducers, in their effects on biologically distinguishable viruses. The use of such inhibitors to study integrated virus expression provides a means for studying gene regulation in mammalian cells.


Subdural electrodes in focal epilepsy surgery at a typical academic epilepsy center.
To assess the use of subdural intracranial EEG (iEEG) on postoperative outcomes at an epilepsy center. Ninety-one epilepsy patients underwent iEEG. Array design was compared with noninvasive EEG with over 1 year of outcome. Patient demographics, implanted brain sites, electrodes, contacts/site, and surgical location were correlated with outcome. Fisher exact test and logistics regression were used to evaluate significance (P ≤ 0.05). Of ninety-one women, 55 (mean age, 32.3 years; range, 11-60) underwent tailored iEEG. Seventy of ninety-one (76.9%) resections (70% temporal) yielded 24/91 (26.4%) seizure free (SF). Strips (57.1%), grids (5.5%), or both (37.4%) for iEEG use was commonly bilateral (58.2%; 65.3% bitemporal) but did not predict outcome (P = NS). A lesion (28/91) did predict a SF outcome (42.9%). The iEEG localized 45.7% of seizures beyond scalp EEG and changed the localization or lateralization in 75.7% of resected patients. Electrode design, localization, lateralization, and site of resection did not correlate with outcome (P = NS). Overall, iEEG use portended a non-SF outcome (P ≤ 0.0001). The use of iEEG selected 46% additional patients for surgery, yet only 26% became SF. A magnetic resonance imaging lesion predicted a SF or seizure-improved outcome. Although iEEG changed the localization and lateralization of scalp ictal EEG in three quarters of patients, its use was a negative predictor for a favorable outcome. Preoperative counseling should emphasize expectations for seizure reduction in patients requiring iEEG.


Resting Energy Expenditure in Patients with Stroke during the Subacute Phases - Relationships with Stroke Types, Location, Severity of Paresis, and Activities of Daily Living.
The energy demands in patients with stroke during the subacute phases are unclear. However, this information is essential for appropriate clinical and nutritional management. The aims of this study were to determine the resting energy expenditure (REE) during the subacute phases, examine its relationships with stroke types, location, severity of hemiparesis, and activities of daily living (ADL), and evaluate whether estimation of REE from the Harris-Benedict equation (HB) requires the addition of a 'stress factor' to capture possible additional REE imposed by stroke. We measured REE in 95 patients with subacute stroke (53.5 ± 16.6 days post-stroke) with indirect calorimetry, and compared it with predicted values of energy expenditure estimated from the HB (expressed as percentage). Patients were admitted for rehabilitation of their first ischemic or nonsurgical hemorrhagic stroke. The severity of hemiparesis was assessed with the motor items of the Stroke Impairment Assessment Set (SIAS). The ADL was assessed with the Functional Independence Measure (FIM). We compared REE and %HB of the two groups divided by hemiparesis severity and ADL limitation using the Student's t-test, and those of the five groups divided by stroke location. The correlations between REE and the motor items of the SIAS and the FIM score were assessed with the Spearman rank correlation test. A multiple regression analysis for REE was conducted. The average body weight (BW) was 57.1 ± 11.3 kg. The average body mass index (BMI) was 22.5 ± 4.0. The mean REE (%HB) was 1,271 ± 284 kcal/day (106.0 ± 17.3%). REE and %HB of the low ADL group was less than that of the high ADL group (p < 0.05). The REE had a positive correlation with the FIM score (rs = 0.51, p < 0.01). The motor items of the SIAS were not significantly correlated with REE. BW, FIM, and stroke location were independent predictors of REE. Analysis of energy expenditure suggests that stroke patients are not hypermetabolic during the subacute phase. The 'stress factor' in stroke patients during the subacute phase was 1.0-1.1. This finding provides important information for improving nutritional management during the subacute phase in patients with stroke.


Astrocyte-specific regulation of hMeCP2 expression in Drosophila.
Alterations in the expression of Methyl-CpG-binding protein 2 (MeCP2) either by mutations or gene duplication leads to a wide spectrum of neurodevelopmental disorders including Rett Syndrome and MeCP2 duplication disorder. Common features of Rett Syndrome (RTT), MeCP2 duplication disorder, and neuropsychiatric disorders indicate that even moderate changes in MeCP2 protein levels result in functional and structural cell abnormalities. In this study, we investigated two areas of MeCP2 pathophysiology using Drosophila as a model system: the effects of MeCP2 glial gain-of-function activity on circuits controlling sleep behavior, and the cell-type specific regulation of MeCP2 expression. In this study, we first examined the effects of elevated MeCP2 levels on microcircuits by expressing human MeCP2 (hMeCP2) in astrocytes and distinct subsets of amine neurons including dopamine and octopamine (OA) neurons. Depending on the cell-type, hMeCP2 expression reduced sleep levels, altered daytime/nighttime sleep patterns, and generated sleep maintenance deficits. Second, we identified a 498 base pair region of the MeCP2e2 isoform that is targeted for regulation in distinct subsets of astrocytes. Levels of the full-length hMeCP2e2 and mutant RTT R106W protein decreased in astrocytes in a temporally and spatially regulated manner. In contrast, expression of the deletion Δ166 hMeCP2 protein was not altered in the entire astrocyte population. qPCR experiments revealed a reduction in full-length hMeCP2e2 transcript levels suggesting transgenic hMeCP2 expression is regulated at the transcriptional level. Given the phenotypic complexities that are caused by alterations in MeCP2 levels, our results provide insight into distinct cellular mechanisms that control MeCP2 expression and link microcircuit abnormalities with defined behavioral deficits.


How long do smoke alarms function? A cross-sectional follow-up survey of a smoke alarm installation programme.
To assess the functionality of lithium-powered smoke alarms that had been installed through a community-based programme called Operation Installation (OI). A random sample was chosen of homes that had received smoke alarms through OI, 2, 4, 6, 8 and 10 years previously. Sampled homes were visited, and information collected included functional status of smoke alarms. For homes in the 6-, 8- and 10-year sample, smoke alarms were removed and tested for battery and alarm function. 800 homes were included in the survey results; 1884 smoke alarms had been installed through OI. The proportion of homes that had at least one functioning OI smoke alarm ranged from 91.8% for year 2 sample to 19.8% for year 10. Of the originally installed smoke alarms in year 10 sample, 45.5% had been removed and 59% (64/108) of those that were still installed were not functioning. Multivariate analysis showed that the presence of at least one working alarm in the home was associated positively with the number of smoke alarms that were originally installed and whether the original occupant was still living in the home, and negatively with the length of time since the smoke alarm was installed, and whether there was a smoker in the home. Testing of the smoke alarms revealed that most non-functioning alarms had missing or dead batteries. Less than a quarter of the originally installed smoke alarms were still present and functioning by year 10. These findings have important implications for smoke alarm installation programmes.


The Saccharomyces cerevisiae Cdk8 Mediator Represses AQY1 Transcription by Inhibiting Set1p-Dependent Histone Methylation.
In the budding yeast Saccharomyces cerevisiae, nutrient depletion induces massive transcriptional reprogramming that relies upon communication between transcription factors, post-translational histone modifications, and the RNA polymerase II holoenzyme complex. Histone H3Lys4 methylation (H3Lys4 me), regulated by the Set1p-containing COMPASS methyltransferase complex and Jhd2p demethylase, is one of the most well-studied histone modifications. We previously demonstrated that the RNA polymerase II mediator components cyclin C-Cdk8p inhibit locus-specific H3Lys4 3me independently of Jhd2p Here, we identify loci subject to cyclin C- and Jhd2p-dependent histone H3Lys4 3me inhibition using chromatin immunoprecipitation (ChIP)-seq. We further characterized the independent and combined roles of cyclin C and Jhd2p in controlling H3Lys4 3me and transcription in response to fermentable and nonfermentable carbon at multiple loci. These experiments suggest that H3Lys4 3me alone is insufficient to induce transcription. Interestingly, we identified an unexpected role for cyclin C-Cdk8p in repressing AQY1 transcription, an aquaporin whose expression is normally induced during nutrient deprivation. These experiments, combined with previous work in other labs, support a two-step model in which cyclin C-Cdk8p mediate AQY1 transcriptional repression by stimulating transcription factor proteolysis and preventing Set1p recruitment to the AQY1 locus.


CD25+CD4+ regulatory T cells generated by exposure to a model protein antigen prevent allograft rejection: antigen-specific reactivation in vivo is critical for bystander regulation.
The importance of CD25(+)CD4(+) regulatory T (Treg) cells in the control of immune responses is established, but their antigen specificity in vivo remains unclear. Understanding Treg-cell specificity requirements will be important if their potential is to be developed for immunotherapy. Pretreatment of recipient mice with donor alloantigen plus anti-CD4 antibody generates CD25(+)CD4(+) Treg cells with the capacity to prevent skin allograft rejection in adoptive transfer recipients. Here we demonstrate that, although this regulation can be antigen-specific, reactivation with the original tolerizing alloantigen allows the Treg cells to suppress rejection of third-party allografts. Aware of the limitations of alloantigen pretreatment, we asked whether graft-protective Treg cells could be generated against unrelated, nongraft antigens. We demonstrate that bystander regulation also extends to CD25(+)CD4(+) Treg cells generated in vivo by exposure to nominal antigens under anti-CD4 antibody cover. Providing these Treg cells are reexposed to the tolerizing antigens before adoptive transfer, they prevent the rejection of fully allogeneic skin grafts. That this might form the basis of a clinically relevant tolerance induction strategy is demonstrated by the fact that, when combined with subtherapeutic anti-CD8 antibody, Treg cells generated in response to nongraft antigens facilitate the acceptance of cardiac allografts in primary recipients.


Decompressive surgery for treating nerve damage in leprosy.
Leprosy causes nerve damage which may result in nerve function impairment and disability. Decompressive surgery is used for treating nerve damage, although the effect is uncertain. To assess the effects of decompressive surgery on nerve damage in leprosy. We searched the Cochrane Neuromuscular Disease Group Trials Register (November 2007), the Cochrane Central Register of Controlled Trials (The Cochrane Library Issue 4, 2007), MEDLINE (from January 1950 to November 2007), EMBASE (from January 1980 to November 2007), AMED (from January 1985 to November 2007), CINAHL (from January 1982 to November 2007) and LILACS (from January 1982 to November 2007) in November 2007. We checked reference lists of the studies identified, the Current Controlled Trials Register (www.controlled-trials.com), conference proceedings and contacted trial authors. Randomised and quasi-randomised controlled trials of decompressive surgery for nerve damage in leprosy. The primary outcome was improvement in sensory and motor nerve function after one year. Secondary outcomes were improvement in nerve function after two years, change in nerve pain and tenderness, and adverse events. Two authors independently extracted data and assessed trial quality. We contacted trial authors for additional information. We collected adverse effects information from the trials and non-randomised studies. We included two randomised controlled trials involving 88 people. The trials examined the added benefit of surgery over prednisolone for treatment of nerve damage of less than six months duration. After two years follow-up there was no significant difference in nerve function improvement between people treated with surgery plus prednisolone or with prednisolone alone. Adverse effects of decompressive surgery were not adequately described. Decompressive surgery is used for treating nerve damage in leprosy but evidence from randomised controlled trials does not show a significant added benefit of surgery over steroid treatment alone. Well-designed randomised controlled trials are needed to establish the effectiveness of the combination of surgery and medical treatment compared to medical treatment alone.


Activation-induced cytidine deaminase targets DNA at sites of RNA polymerase II stalling by interaction with Spt5.
Activation-induced cytidine deaminase (AID) initiates antibody gene diversification by creating U:G mismatches. However, AID is not specific for antibody genes; Off-target lesions can activate oncogenes or cause chromosome translocations. Despite its importance in these transactions little is known about how AID finds its targets. We performed an shRNA screen to identify factors required for class switch recombination (CSR) of antibody loci. We found that Spt5, a factor associated with stalled RNA polymerase II (Pol II) and single stranded DNA (ssDNA), is required for CSR. Spt5 interacts with AID, it facilitates association between AID and Pol II, and AID recruitment to its Ig and non-Ig targets. ChIP-seq experiments reveal that Spt5 colocalizes with AID and stalled Pol II. Further, Spt5 accumulation at sites of Pol II stalling is predictive of AID-induced mutation. We propose that AID is targeted to sites of Pol II stalling in part via its association with Spt5.