The Bet Tzedek legal services model: how a legal services model addresses elder abuse and neglect.
Bet Tzedek, Hebrew for the "House of Justice," provides free legal assistance to older adults in Los Angeles County. Their civil attorneys work alongside prosecutors and service providers for the elderly as members of multidisciplinary teams to assist older adults with complicated elder abuse and neglect cases. Case examples demonstrate how civil attorneys collaborate with the Los Angeles County Elder Abuse Forensic Center to address financial abuse, real estate fraud, and self-neglect issues. Cooperation among the courts, Bet Tzedek, and other county agencies has resulted in more user-friendly processes to expedite filing of conservatorships and elder abuse restraining orders.


[Treatment of intact myometrial cells with oxytocin inhibits Mg2+, ATP-dependent accumulation of Ca2+ in endoplasmic reticulum].
Uterotonic peptide hormone oxytocin has been studied for its effect on ATP-dependent accumulation of Ca2+ nonsensitive to the effect of ruthenium red (10 microM) that is potentiated by Ca2+-precipitating anion - oxalate. That was studied in experiments made on the suspension of myometrium cells from estrogenized rats processed by digitonin (0.1 mg/ml) with the aim permeabilization of plasmatic membrane. The preliminary processing of intact myocytes by oxytocin solution (final concentration 100 mM) to permeabilization of plasmatic membrane causes partial inhibition (stimulated by oxalate by 25-30% (0-10 mM) of Mg2+, ATP-dependent accumulation of Ca2+ in nonmitochondrion intracellular calcium depot of perforates smooth-muscled cells. The value of the component of power-dependent accumulation of Ca2+, that is inhibited by oxytocin, increased with both the incubation time and oxalate concentration. Thapsigargin and cyclopiasonium acid, selective inhibitors of calcium pump of endo(sarco)plasmatic reticulum, when used in saturation concentration (50 mM and 10 mM, respectively), completely inhibit the component of Mg2+, ATP-dependent accumulation of Ca2+ in nonmitochondrial intracellular calcium depot that is stimulated with oxalate (< 10 mM) and inhibited with oxytocin (100 mM). It is concluded that oxytocin partially inhibits Mg2+, ATP-dependent calcium pump of myometrium cell endoplasmic reticulum.


Three dimensional high-performance micro-supercapacitors with switchable high power density and high energy density.
In the field of microscale energy storage, the fabrication of micro-supercapacitors (MSCs) with high power density and high energy density has always been a focus of research. In this work, laser-induced porous graphene and chemically deposited manganese dioxide nanoparticles are used as electrode materials, and a switchable MSC with two energy storage principles is obtained by designing symmetric interdigitated and square electrode structures. The aim is to overcome the preparation challenge of supercapacitors with high energy density and high power density by switching between two modes. In this MSC, the energy density of the high energy density mode (5.89 μW h cm-2) is 3.36 times that of the high power density mode (1.75 μW h cm-2), while the power density of the high power density mode (43.06 μW cm-2) is 1.44 times that of the high energy density mode (29.96 μW cm-2). In addition, under the drive of five serially connected MSCs, 27 LED lights can be continuously lit for 5 minutes. Therefore, this work provides a facile and novel method for the development of MSCs with high power density and high energy density, suggesting a great practical application value in the development of MSCs.


Single-Particle Enzyme Activity Assay with Spectral-Resolved Dark-Field Optical Microscopy.
In a clinical assay, enzymes are essential biomarkers for human disease diagnosis. In this work, a spectral-resolved single-particle detection (SPD) method is introduced to quantify alkaline phosphatase (ALP) activity in human serum with a supraparticle (SP) based on MnO2-modified gold nanoparticle (denoted as GNP@MnO2 SP) as the probe. In the presence of ALP, 2-phospho-l-ascorbic acid trisodium salt can be hydrolyzed into l-ascorbic acid, which serves as a good reduction agent to trigger the decomposition of the MnO2 shell on the GNP surface. Given that a trace amount of ALP exists, noticeable scattering color change can be detected at the single-particle level due to the sensitive localized surface plasmon resonance (LSPR) effect from GNPs. With spectral-resolved dark-field optical microscopy, a linear dynamic range of 0.06 to 2.48 mU/mL ( R2 = 0.99) and a very low limit of detection of 5.8 μU/mL for the ALP assay are readily achieved, which is more sensitive over the methods based on ensemble sample measurement. As a consequence, this strategy opens a new avenue for the design of an ultrasensitive detection method for disease-correlated biomarker diagnosis in the future.


Assessment of spatial normalization of PET ligand images using ligand-specific templates.
Recent advances allow robust computation of parametric maps of ligand-receptor binding from PET data sets. Parametric maps may be statistically analyzed at the voxel level, given suitable techniques for both the spatial normalization of image data into a standard space and the application of appropriate statistical tests. The purpose of this study was to spatially normalize parametric maps of [carbonyl-11C]WAY-100635 and [11C]raclopride binding using SPM 96 and ligand-specific templates. Ligand-specific templates were created from integral images taken from healthy subjects. For this, a MRI-based spatial normalization was used: T1-weighted MRI scans were coregistered to the PET integral images, and the spatial normalization of the MRI to the SPM 96 T1 MRI template was applied to the integral images. These integral images were meaned and smoothed to form [carbonyl-11C]WAY-100635 and [11C]raclopride templates. Reliability of spatial normalization using the ligand template method and the previous MRI-based spatial normalization was investigated by using a second set of integral images taken from a different cohort: Landmark coordinates were defined on all spatially normalized integral images. Mean coordinates were found in order to produce an overall (average) landmark for each location. For each image, at each location, the distance from the landmark coordinates to the overall landmark were found. A multivariate analysis of variance was used to examine the effects of observer variance, landmark location, and the method used. Visually acceptable templates were created. While observer variance was not significant, the landmark x method interaction was significant. The ligand template method had significantly smaller distances: Among the landmark locations with this method, the mean distances between individual image landmarks and overall image landmarks ranged from 1. 1 to 4.9 mm. The ligand template method provides a reliable approach for spatial normalization of PET ligand images.


Haptic wearables as sensory replacement, sensory augmentation and trainer - a review.
Sensory impairments decrease quality of life and can slow or hinder rehabilitation. Small, computationally powerful electronics have enabled the recent development of wearable systems aimed to improve function for individuals with sensory impairments. The purpose of this review is to synthesize current haptic wearable research for clinical applications involving sensory impairments. We define haptic wearables as untethered, ungrounded body worn devices that interact with skin directly or through clothing and can be used in natural environments outside a laboratory. Results of this review are categorized by degree of sensory impairment. Total impairment, such as in an amputee, blind, or deaf individual, involves haptics acting as sensory replacement; partial impairment, as is common in rehabilitation, involves haptics as sensory augmentation; and no impairment involves haptics as trainer. This review found that wearable haptic devices improved function for a variety of clinical applications including: rehabilitation, prosthetics, vestibular loss, osteoarthritis, vision loss and hearing loss. Future haptic wearables development should focus on clinical needs, intuitive and multimodal haptic displays, low energy demands, and biomechanical compliance for long-term usage.


Lipoxins in the Nervous System: Brighter Prospects for Neuroprotection.
Lipoxins (LXs) are generated from arachidonic acid and are involved in the resolution of inflammation and confer protection in a variety of pathological processes. In the nervous system, LXs exert an array of protective effects against neurological diseases, including ischemic or hemorrhagic stroke, neonatal hypoxia-ischemia encephalopathy, brain and spinal cord injury, Alzheimer's disease, multiple sclerosis, and neuropathic pain. Lipoxin administration is a potential therapeutic strategy in neurological diseases due to its notable efficiency and unique superiority regarding safety. Here, we provide an overview of LXs in terms of their synthesis, signaling pathways and neuroprotective evidence. Overall, we believe that, along with advances in lipoxin-related drug design, LXs will bring brighter prospects for neuroprotection.


1-O-Acetyl-beta-D-galactopyranose: a novel substrate for the transglycosylation reaction catalyzed by the beta-galactosidase from Penicillium sp.
1-O-Acetyl-beta-D-galactopyranose (AcGal), a new substrate for beta-galactosidase, was synthesized in a stereoselective manner by the trichloroacetimidate procedure. Kinetic parameters (K(M) and k(cat)) for the hydrolysis of 1-O-acetyl-beta-D-galactopyranose catalyzed by the beta-D-galactosidase from Penicillium sp. were compared with similar characteristics for a number of natural and synthetic substrates. The value for k(cat) in the hydrolysis of AcGal was three orders of magnitude greater than for other known substrates. The beta-galactosidase hydrolyzes AcGal with retention of anomeric configuration. The transglycosylation activity of the beta-D-galactosidase in the reaction of AcGal and methyl beta-D-galactopyranoside (1) as substrates was investigated by 1H NMR spectroscopy and HPLC techniques. The transglycosylation product using AcGal as a substrate was beta-D-galactopyranosyl-(1-->6)-1-O-acetyl-beta-D-galactopyranose (with a yield of approximately 70%). In the case of 1 as a substrate, the main transglycosylation product was methyl beta-D-galactopyranosyl-(1-->6)-beta-D-galactopyranoside. Methyl beta-D-galactopyranosyl-(1-->3)-beta-D-galactopyranoside was found to be minor product in the latter reaction.


Legionella pneumophila-Derived Outer Membrane Vesicles Promote Bacterial Replication in Macrophages.
The formation and release of outer membrane vesicles (OMVs) is a phenomenon of Gram-negative bacteria. This includes Legionella pneumophila (L. pneumophila), a causative agent of severe pneumonia. Upon its transmission into the lung, L. pneumophila primarily infects and replicates within macrophages. Here, we analyzed the influence of L. pneumophila OMVs on macrophages. To this end, differentiated THP-1 cells were incubated with increasing doses of Legionella OMVs, leading to a TLR2-dependent classical activation of macrophages with the release of pro-inflammatory cytokines. Inhibition of TLR2 and NF-κB signaling reduced the induction of pro-inflammatory cytokines. Furthermore, treatment of THP-1 cells with OMVs prior to infection reduced replication of L. pneumophila in THP-1 cells. Blocking of TLR2 activation or heat denaturation of OMVs restored bacterial replication in the first 24 h of infection. With prolonged infection-time, OMV pre-treated macrophages became more permissive for bacterial replication than untreated cells and showed increased numbers of Legionella-containing vacuoles and reduced pro-inflammatory cytokine induction. Additionally, miRNA-146a was found to be transcriptionally induced by OMVs and to facilitate bacterial replication. Accordingly, IRAK-1, one of miRNA-146a's targets, showed prolonged activation-dependent degradation, which rendered THP-1 cells more permissive for Legionella replication. In conclusion, L. pneumophila OMVs are initially potent pro-inflammatory stimulators of macrophages, acting via TLR2, IRAK-1, and NF-κB, while at later time points, OMVs facilitate L. pneumophila replication by miR-146a-dependent IRAK-1 suppression. OMVs might thereby promote spreading of L. pneumophila in the host.


Contemporary transsphenoidal surgery for pituitary adenomas with emphasis on complications.
The indication and complications of transsphenoidal surgery (TSS) were reported with an emphasis on our experiences. Although new therapies have been developed one after another, TSS is still main stream in the treatment of most pituitary adenomas. One century after the birth of TSS, it has evolved into a more sophisticated and less invasive procedure. The present microscopic TSS will be replaced by endoscopic TSS in the near future. Complications can occur with any surgery, but we can reduce their possibility and degree by enriching experience. In our series, the morbidity rate was 2.7%, but operative death was absent. A better understanding of the reported complications and a careful preoperative design will contribute to a better outcome of TSS.


High performance MoS2-based field-effect transistor enabled by hydrazine doping.
We investigated the n-type doping effect of hydrazine on the electrical characteristics of a molybdenum disulphide (MoS2)-based field-effect transistor (FET). The threshold voltage of the MoS2 FET shifted towards more negative values (from -20 to -70 V) on treating with 100% hydrazine solution with the channel current increasing from 0.5 to 25 μA at zero gate bias. The inverse subthreshold slope decreased sharply on doping, while the ON/OFF ratio increased by a factor of 100. Gate-channel coupling improved with doping, which facilitates the reduction of channel length between the source and drain electrodes without compromising on the transistor performance, making the MoS2-based FET easily scalable.


Toward the complete prediction of the 1H and 13C NMR spectra of complex organic molecules by DFT methods: application to natural substances.
The NMR parameters (1H and 13C chemical shifts and coupling constants) for a series of naturally occurring molecules have been calculated mostly with DFT methods, and their spectra compared with available experimental ones. The comparison includes strychnine as a test case, as well as some examples of recently isolated natural products (corianlactone, daphnipaxinin, boletunone B) featuring unusual and/or crowded structures and, in the case of boletunone B, being the subject of a recent revision. Whenever experimental spectra were obtained in polar solvents, the calculation of NMR parameters was also carried out with the Integral Equation-Formalism Polarizable Continuum Model (IEF-PCM) continuum method. The computed results generally show a good agreement with experiment, as judged not only by statistical parameters but also by visual comparison of line spectra. The origin of the remaining discrepancies is attributed to the incomplete modeling of conformational and specific solvent effects.


Endoscopic Endonasal Repair of the Orbital Medial Wall Fracture Utilising Nasal Septal Cartilage as the Reconstructive Material.
To repair orbital medial wall fractures, otorhinolaryngologists often use the silicone sheet technique by inserting an inverted U-shaped silicone sheet into the middle meatus after the removal of fractured bones, and packing gauze inside the silicone sheet for several days to fix the shape of the medial wall. However, this method does not sufficiently reduce the orbital content. To describe the surgical procedure to repair medial wall fractures using nasal septal cartilage as the reconstructive material. First, endoscopic septoplasty is performed. Although the correction of the septal deviation is performed to secure the operative field, the nasal septal cartilage, except the L-strut is resected simultaneously. After septoplasty, the fractured bones of the lamina papyracea are removed from the herniated orbital contents, and then the orbital contents can be reduced without resistance by pushing laterally. Subsequently, the cartilage is inserted beneath the fracture edges of the orbital medial wall with great care to avoid entrapping the orbital soft tissue and extraocular muscles. The cartilage can be placed appropriately along the medial wall of the orbit. This technique enables to repair most cases of orbital medial wall fractures using the endoscopic endonasal approach alone without alloplastic implants.


Long-term results of tension-free vaginal tape (TVT) for the treatment of female urinary stress incontinence.
Prospective evaluation of outcome and complications over a 5-year period post-treatment of urinary stress incontinence by TVT, and comparison of our results with the reference studies. About 94 patients were treated for urinary stress incontinence only by one TVT procedure (single surgical procedure), between April 1997 and December 1998; 68% of patients presented pure urinary stress incontinence and 32% mixed incontinence. We found also a 25.5% rate of sphincter deficiency (UCP < 20 cm H(2)O) in this cohort. Patients were evaluated after 5 years: 52 complete evaluations (clinical, flow measurement with measurement of post-mictional residue, 24h PAD-test, quality of life questionnaire), 30 complete telephone interviews, 12 lost to follow-up (2 patients deceased). About 87% of the patients had a 5-year follow-up. The success rate was 79.2% overall (84.5% for the pure urinary stress incontinence and 67% for the mixed incontinence cases), and 72.2% for the cases of associated sphincter deficiency. We had only a 13% rate of patients lost to follow-up. More than half of the urinary urgency cases were treated successfully, however with a less satisfactory outcome in cases of bladder instability. The urodynamic exploration appeared to reveal that TVT caused dysuria: 52% of patients had a maximum flowrate below 15 ml/s, but the quality of life was improved, with a 95% rate of satisfaction without functional problems. We observed no late complications such as vaginal erosion or rejection of the prolene; the de novo syndrome was rare, with 8.5% of urinary frequency, 6% of urinary urgency and only 5.7% of invalidating dysuria. We saw no cases of pelvic floor disease after TVT treatment. Our casuistry results are comparable with the reference studies by Scandinavian authors, Rezapour and Ulmsten, confirming the long-term success of the TVT procedure. Concerning the apparently elevated rates of post-TVT dysuria found by urodynamic exploration, a distinction has to be drawn between post-TVT urinary problems (frequent but oligosymptomatic), and true, severe dysuria (rare). However, "dysuria" in the broad sense did not affect the patients' quality of life, and is a reminder of the absolute necessity of meticulous compliance with the correct surgical techniques. Treatment of urinary incontinence by TVT is a reliable, mini-invasive, reproducible technique, almost suitable for outpatients, with no serious complications; it is inexpensive and very successful, including in complicated cases such as sphincter deficiency. All the recent data confirms, with this 5-year follow-up, that the TVT procedure is comparable to the previously gold standard, the Burch colposuspension.


The deep roots of military service in neurological surgery: an academic genealogical analysis of the founding generation.
Throughout human history, advancements in medicine have evolved out of periods of war. The carnage of battlefield injuries provided wartime surgeons an unprecedented opportunity to study anatomy, develop novel techniques, and improve systems of care. As a specialty that was established and evolved during the first half of the 20th century, neurological surgery was heavily influenced by the experiences of its founders during the World Wars I and II. Utilizing the published Neurosurgery Tree, the authors conducted an academic genealogical analysis to systematically define the influence of wartime service on neurosurgery's earliest generations. Through review of the literature and military records, the authors determined that at least 60% of American neurosurgical founders and early leaders served during World Wars I and/or II. Inspired by the call to serve their nation as forces for good, these individuals were heralded as expert clinicians, innovative systems thinkers, and prolific researchers. Importantly, the service of these early leaders helped highlight the viability of neurosurgery as a distinct specialty and provided a framework for early neurosurgical education and expansion. The equipment, techniques, and guidelines that were developed during these wars, such as management of craniocerebral trauma, peripheral nerve repair, and hemostasis, set the foundation for modern neurosurgical practice.


Regulation of WNK1 expression by miR-192 and aldosterone.
WNK1 and WNK4 encode two members of the WNK serine-threonine kinase subfamily. Greater WNK1 expression associates with higher BP. A combination of promoters, enhancers, repressors, and insulators regulate WNK1 expression, but whether microRNAs also modulate WNK1 expression is unknown. Here, computational analysis revealed the presence of a target sequence for miR-192 and miR-215 at the same site in the 3' untranslated region of the ubiquitous L- and the kidney-specific KS-WNK1. We functionally validated this target sequence by transient transfection and reporter assays. Although we observed expression of both miRs along the distal nephron, only miR-192 regulated endogenous WNK1 ex vivo. Furthermore, a potassium load, sodium depletion, and aldosterone infusion each significantly reduced miR-192 expression in the kidney. Taken together, these results suggest a miR-driven mechanism of gene regulation by aldosterone and a role for miR-192 in the regulation of sodium and potassium balance in the kidney.


Implementing an EMR-Based Referral for Smoking Quitline Services with Additional Provider Education, a Cluster-Randomized Trial.
Despite evidence of their effectiveness, free smoking quitlines are underused. The best way to educate providers about and encourage use of quitlines is not established. We examined if electronic medical record (EMR)-integrated best practices alerts (BPAs) with or without additional provider education resulted in increased quitline referrals. Waitlist-controlled, cluster-randomized trial of primary care practices assigned to three arms. Providers in participating sites received a new EMR-based BPA for quitline referral and additional education outreach visits, the BPA alone, or usual care. The study was conducted in 2 phases: phase 1 from April 17 to October 16, 2017, and phase 2 from November 9, 2017, to May 8, 2018. In phase 2, the usual-care sites were randomized to either of the two intervention arms. The unit of randomization was primary care practice site. All in-office, primary care provider visits with smokers were included. The primary outcome was referral to the quitline. Secondary outcomes included patient acceptance and enrollment in quitline services. Twenty-two practice sites were enrolled. Smoking prevalence at sites ranged from 4.4 to 23%. In phase 1, the BPA-plus-education arm had 5636 eligible encounters and 405 referrals (referral rate 7.2%) while the BPA-only arm had 6857 eligible encounters and 623 referrals (referral rate 9.1%). The usual-care arm had 7434 encounters but no referrals. Comparing the BPA-plus arm to the BPA-only arm, the odds ratio of referral was 0.76 (CI 0.3-1.8). In phase 2, the combined BPA-plus-education sites had 8516 eligible encounters and 475 referrals (rate 5.6%). The BPA-only sites had 9134 eligible encounters and 470 referrals (rate 5.2%). The odds ratio comparing the 2 groups in phase 2 was 1.06 (0.5-2.2). An EMR-based BPA can improve the number of referrals to quitline services, though more work is needed to improve providers' use of quitlines and low patient acceptance of services. Trial Registration NIH Clinicaltrials.gov identifier: NCT03229356.


Diagnostic results before and after introduction of autofluorescence bronchoscopy in patients suspected of having lung cancer detected by sputum cytology in lung cancer mass screening.
For the purpose of early detection, we have conducted population-based mass screening for lung cancer by sputum cytology since 1982. Although detection of lung cancer in its early stage is important for a good prognosis, it is often difficult to localize lesions in roentgenographically occult cancer. To clarify the role of autofluorescence bronchoscopy in localizing tumors in patients with roentgenographically occult cancer, we analyzed our diagnostic results. Fifty patients who had been detected by sputum cytology were screened by the light-induced fluorescence endoscope (LIFE)-Lung System from November 1997 to April 1999. We compared the results according to the screening methods: conventional bronchoscopy alone versus LIFE with conventional white-light bronchoscopy (November 1997 to April 1999). Twenty-eight cancerous lesions and 39 borderline lesions were detected by LIFE. Of the 39 borderline lesions, nine were detected only by LIFE. Multicentric lesions including cancer or dysplasia were also detected in 21 of the 50 patients by LIFE. The sensitivity by white-light bronchoscopy alone was 85.3%, whereas that of the LIFE-Lung System with white-light bronchoscopy was 94.1% (P=0.078). There were no cancerous lesions in the area observed as normal by LIFE. We also compared the diagnostic results of two localization methods: brushing of all bronchi (September 1986 to December 1990) and the LIFE-Lung System (November 1997 to April 1999). Although this was a historical comparison, the number of detected borderline lesions increased, which led to a high detection rate in patients with suspected-positive sputum (P=0.0006) by the LIFE-Lung System. In conclusion, the LIFE-Lung System is a safe and non-invasive system for detecting small intraepithelial lesions of the tracheobronchial tree. Autofluorescence bronchoscopy is more efficacious for localizing intraepithelial lesions and places fewer burdens on the patient than brushing of all bronchi.


Treating From the Inside Out: Relevance of Fecal Microbiota Transplantation to Counteract Gut Damage in GVHD and HIV Infection.
The gastrointestinal (GI) tract is a complex and well-balanced milieu of anatomic and immunological barriers. The epithelial surface of the GI tract is colonized by trillions of microorganisms, known as the gut microbiota, which is considered an "organ" with distinctive endocrine and immunoregulatory functions. Dysregulation of the gut microbiota composition, termed dysbiosis, has been associated with epithelial damage and translocation of microbial products into the circulating blood. Dysbiosis, increased gut permeability and chronic inflammation play a major role on the clinical outcome of inflammatory bowel diseases, graft-vs.-host disease (GVHD) and HIV infection. In this review, we focus on GVHD and HIV infection, conditions sharing gut immune damage leading to dysbiosis. The degree of dysbiosis and level of epithelial gut damage predict poor clinical outcome in both conditions. Emerging interventions are therefore warranted to promote gut microbiota homeostasis and improve intestinal barrier function. Interventions such as anti-inflammatory medications, and probiotics have toxicity and/or limited transitory effects, justifying innovative approaches. Fecal microbiota transplantation (FMT) is one such approach where fecal microorganisms are transferred from healthy donors into the GI tract of the recipient to restore microbiota composition in patients with Clostridium difficile-induced colitis or inflammatory bowel diseases. Preliminary findings point toward a beneficial effect of FMT to improve GVHD and HIV-related outcomes through the engraftment of beneficial donor bacteria, notably those producing anti-inflammatory metabolites. Herein, we critically review the potential for FMT in alleviating dysbiosis and gut damage in patients with GVHD or HIV-infection. Understanding the underlying mechanism by which FMT restores gut function will pave the way toward novel scalable and targeted interventions.


Systematic Standardized and Individualized Assessment of Masticatory Cycles Using Electromagnetic 3D Articulography and Computer Scripts.
Masticatory movements are studied for decades in odontology; a better understanding of them could improve dental treatments. The aim of this study was to describe an innovative, accurate, and systematic method of analyzing masticatory cycles, generating comparable quantitative data. The masticatory cycles of 5 volunteers (Class I, 19 ± 1.7 years) without articular or dental occlusion problems were evaluated using 3D electromagnetic articulography supported by MATLAB software. The method allows the trajectory morphology of the set of chewing cycles to be analyzed from different views and angles. It was also possible to individualize the trajectory of each cycle providing accurate quantitative data, such as number of cycles, cycle areas in frontal view, and the ratio between each cycle area and the frontal mandibular border movement area. There was a moderate negative correlation (-0.61) between the area and the number of cycles: the greater the cycle area, the smaller the number of repetitions. Finally it was possible to evaluate the area of the cycles through time, which did not reveal a standardized behavior. The proposed method provided reproducible, intelligible, and accurate quantitative and graphical data, suggesting that it is promising and may be applied in different clinical situations and treatments.


Molecular evolution of P transposable elements in the genus Drosophila. I. The saltans and willistoni species groups.
A phylogenetic survey using the polymerase chain reaction (PCR) has identified four major P element subfamilies in the saltans and willistoni species groups of Drosophila. One subfamily, containing about half of the sequences studied, consists of elements that are very similar to the canonical (and active) P element from D. melanogaster. Within this subfamily, nucleotide sequence differentiation among different copies from the same species and among elements from different species is relatively low. This observation suggests that the canonical elements are relatively recent additions to the genome or, less likely, are evolving slowly relative to the other subfamilies. Elements belonging to the three noncanonical lineages are distinct from the canonical elements and from one another. Furthermore, there is considerably more sequence variation, on the average, within the noncanonical subfamilies compared to the canonical elements. Horizontal transfer and the coexistence of multiple, independently evolving element subfamilies in the same genome may explain the distribution of P elements in the saltans and willistoni species groups. Such explanations are not mutually exclusive, and each may be involved to varying degrees in the maintenance of P elements in natural populations of Drosophila.


Magnetic resonance imaging of rheumatoid arthritis: the evolution of clinical applications through clinical trials.
Powerful techniques are being developed for evaluating rheumatoid arthritis with magnetic resonance imaging (MRI). Much of this development is being driven by the pharmaceutical and biotechnology industries searching for novel therapies for this disease. Accordingly, the imaging tools that ultimately will be used to direct patients to specific therapies and then to monitor treatment effectiveness and safety are currently being refined and validated in rigorous multicenter and multinational clinical trials aimed at gaining regulatory approval of these new therapies. As these trials approach completion, rheumatologists can anticipate an increased demand for expertise and experience in evaluating disease progression and treatment response with these techniques and the emergence of MRI systems specifically designed for this market. The following discussion reviews this novel pathway for evolving imaging techniques for clinical use through clinical drug trials, lists the most promising MRI markers available today for evaluating joint destruction in rheumatoid arthritis, and speculates on how these techniques will find their way into clinical practice.


Epsilon-Near-Zero Grids for On-chip Quantum Networks.
Realization of an on-chip quantum network is a major goal in the field of integrated quantum photonics. A typical network scalable on-chip demands optical integration of single photon sources, optical circuitry and detectors for routing and processing of quantum information. Current solutions either notoriously experience considerable decoherence or suffer from extended footprint dimensions limiting their on-chip scaling. Here we propose and numerically demonstrate a robust on-chip network based on an epsilon-near-zero (ENZ) material, whose dielectric function has the real part close to zero. We show that ENZ materials strongly protect quantum information against decoherence and losses during its propagation in the dense network. As an example, we model a feasible implementation of an ENZ network and demonstrate that information can be reliably sent across a titanium nitride grid with a coherence length of 434 nm, operating at room temperature, which is more than 40 times larger than state-of-the-art plasmonic analogs. Our results facilitate practical realization of large multi-node quantum photonic networks and circuits on-a-chip.


The carbon monoxide derivative of human hemoglobin carrying the double mutation LeuB10-->Tyr and HisE7-->Gln on alpha and beta chains probed by infrared spectroscopy.
The fine structural properties of the distal heme pocket have been probed by infrared spectroscopy of ferrous carbon monoxy human hemoglobin mutants carrying the mutations LeuB10-->Tyr and HisE7-->Gln on the alpha, beta, and both chains, respectively. The stretching frequency of iron-bound carbon monoxide occurs as a single broad band around 1943 cm(-1) in both the alpha and the beta mutated chains. Such a frequency value indicates that no direct hydrogen bonding exists between the bound CO molecule and the TyrB10 phenolic oxygen, at variance with other naturally occurring TyrB10, GlnE7 nonvertebrate hemoglobins. The rates of carbon monoxide release have been determined for the first time by a Fourier transform infrared spectroscopy stopped-flow technique that allowed us to single out the heterogeneity in the kinetics of CO release in the alpha and beta chains for the mutated proteins and for native HbA. The rates of CO release are 15- to 20-fold faster for the mutated alpha or beta chains with respect to the native ones consistent with the lack of distal stabilization on the iron-bound CO molecule. The present results demonstrate that residues in key topological positions (namely E7 and B10) for the distal steric control of the iron-bound ligand are not interchangeable among hemoglobins from different species.


Outcomes and quality of life after platelet-rich plasma therapy in patients with recalcitrant hindfoot and ankle diseases: a preliminary report of 12 patients.
The present study aimed to determine the outcomes and quality of life after platelet-rich plasma therapy in patients with chronic recalcitrant diseases of the hindfoot and ankle and to identify the crucial clinical variables. The records of 12 adult patients with diseases of the hindfoot and ankle were included in the present study. These patients had been treated with platelet-rich plasma from September 2010 to April 2011 after 3 to 6 months or more of conservative treatment had been unsuccessful. They had attended the follow-up visits, were consecutively enrolled, and retrospectively studied. A total of 3 mL of autologous platelet-rich plasma was injected under fluoroscopic or ultrasound guidance into the affected areas. All patients had been evaluated using visual analog scale foot and ankle scoring before treatment and at set intervals after treatment. According to their scores at the final follow-up visit (mean 16 months), the patients were allocated to the satisfactory (score ≥ 80; n = 8) and unsatisfactory (score < 80; n = 4) groups. The health-related quality of life was assessed using the Medical Outcomes Study short-form, 36-item survey at the final follow-up visit, because the study was retrospective, and the information was not available before treatment. The mean visual analog score at the final follow-up visit (79.71 ± 17.81) was significantly greater than the mean pretreatment score (57.89 ± 20.77; p = .002). Four patients (33%) had unsatisfactory results. The mean short-form, 36-item score for the satisfactory group (85.23 ± 11.30) was significantly greater than that (57.33 ± 12.91) of the unsatisfactory group (p = .003). No definitive factors influencing the outcome of this treatment were found. The substantial number of patients with an unsatisfactory outcome indicates that platelet-rich plasma injection might be an option but might not be a mainstay of nonoperative treatment of problematic conditions of the hindfoot and ankle. The actual benefit of this treatment, including the factors influencing its outcome, are still inconclusive.


Chronic recurrent subarachnoid hemorrhage from a trigeminal nerve malignant peripheral nerve sheath tumor: case report.
Clinically significant intratumoral or peritumoral bleeding from trigeminal nerve tumors is very rare. We report the case of a 59-year-old man who presented with recurrent subarachnoid hemorrhage from a left trigeminal nerve malignant peripheral nerve sheath tumor. He presented with decreased consciousness, left facial hypesthesia, and left facial weakness. Trigeminal neuralgia was present for 18 months. Cranial computed tomographic and magnetic resonance imaging scans revealed a left parapontine mass with cystic changes and intratumoral bleeding. Furthermore, signs of hemosiderosis of the subarachnoid space were noted. Lumbar puncture revealed fresh bleeding. Angiography detected no aneurysm or other causes of bleeding. The patient became fully alert within hours, the facial weakness improved within a few days. There was no evidence of vasospasm or persisting hydrocephalus. He underwent left-sided suboccipital craniotomy for macroscopically total tumor removal. The patient underwent total tumor removal via a left suboccipital approach. Intraoperatively, evidence of recurrent intratumoral bleeding was noted. Histological examination revealed a malignant peripheral nerve sheath tumor (World Health Organization Grade III). Postoperatively, the hypesthesia improved significantly. The patient was transferred to radiotherapy for external beam radiation. This is the first report regarding a malignant peripheral nerve sheath tumor of the trigeminal nerve that caused clinically significant subarachnoid hemorrhage caused by intratumoral bleeding.


Interrelationship of magnesium and estrogen in cardiovascular and bone disorders, eclampsia, migraine and premenstrual syndrome.
The anticonvulsive and antihypertensive values of magnesium (Mg) in eclampsia, and its antiarrhythmic applications in a variety of cardiac diseases, have caused Mg to be considered only for parenteral administration by many physicians. In contrast, nutritionists have long recognized Mg as an essential nutrient, because severe deficiencies elicit neuromuscular manifestations similar to those justifying its use in eclampsia. More recently, this element has been used to favorably influence latent tetany with and without thrombotic complications, to delay preterm birth, to influence premenstrual syndrome, and to ameliorate migraine headaches. Most of these disorders exclusively or largely afflict women. The lesions of arteries and heart caused by experimental Mg deficiency have been well documented and may contribute to human cardiovascular disease. Estrogen's enhancement of Mg utilization and uptake by soft tissues and bone may explain resistance of young women to heart disease and osteoporosis, as well as increased prevalence of these diseases when estrogen secretion ceases. However, estrogen-induced shifts of Mg can be deleterious when estrogen levels are high and Mg intake is suboptimal. The resultant lowering of blood Mg can increase the Ca/Mg ratio, thus favoring coagulation. With Ca supplementation in the face of commonly low Mg intake, risk of thrombosis increases.


Linker Molecules Convert Commercial Fluorophores into Tailored Functional Probes during Biolabelling.
Many life-science techniques and assays rely on selective labeling of biological target structures with commercial fluorophores that have specific yet invariant properties. Consequently, a fluorophore (or dye) is only useful for a limited range of applications, e.g., as a label for cellular compartments, super-resolution imaging, DNA sequencing or for a specific biomedical assay. Modifications of fluorophores with the goal to alter their bioconjugation chemistry, photophysical or functional properties typically require complex synthesis schemes. We here introduce a general strategy that allows to customize these properties during biolabelling with the goal to introduce the fluorophore in the last step of biolabelling. For this, we present the design and synthesis of 'linker' compounds, that bridge biotarget, fluorophore and a functional moiety via well-established labeling protocols. Linker molecules were synthesized via the Ugi four-component reaction (Ugi-4CR) which facilitates a modular design of linkers with diverse functional properties and bioconjugation- and fluorophore attachment moieties. To demonstrate the possibilities of different linkers experimentally, we characterized the ability of commercial fluorophores from the classes of cyanines, rhodamines, carbopyronines and silicon-rhodamines to become functional labels on different biological targets in vitro and in vivo via thiol-maleimide chemistry. With our strategy, we showed that the same commercial dye can become a photostable self-healing dye or a sensor for bivalent ions subject to the linker used. Finally, we quantified the photophysical performance of different self-healing linker-fluorophore conjugates and demonstrated their applications in super-resolution imaging and single-molecule spectroscopy.


Earlier Provision of Gastric Bypass Surgery in Canada Enhances Surgical Benefit and Leads to Cost and Comorbidity Reduction.
Background: Effective provision of bariatric surgery for patients with obesity may be impeded by concerns of payers regarding costs or perceptions of patients who drop out of surgical programs after referral. Estimates of the cost and comorbidity impact of these inefficiencies in gastric bypass surgery in Canada are lacking but would aid in informing healthcare investment and resource allocation. Objectives: To estimate total and relative public payer costs for surgery and comorbidities (diabetes, hypertension, and dyslipidemia) in a bariatric surgery population. Methods: A decision analytic model for a 100-patient cohort in Canada (91% female, mean body mass index 49.2 kg/m2, 50% diabetes, 66% hypertension, 59% dyslipidemia). Costs include surgery, surgical complications, and comorbidities over the 10-year post-referral period. Results are calculated as medians and 95% credibility intervals (CrIs) for a pathway with surgery at 1 year ("improved") compared with surgery at 3.5 years ("standard"). Sensitivity analyses were performed to test independent contributions to results of shorter wait time, better post-surgical weight loss, and randomly sampled cohort demographics. Results: Compared to standard care, the improved path was associated with reduction in patient-years of treatment for each of the three comorbidities, corresponding to a reduction of $1.1 (0.68-1.6) million, or 34% (26-41%) of total costs. Comorbidity treatment costs were 9.0- and 4.7-fold greater than surgical costs for the standard and improved pathways, respectively. Relative to non-surgical bariatric care, earlier surgery was associated with earlier return on surgical investment and 2-fold reduction in risk of prevalence of each comorbidity compared to delayed surgery. Conclusions: Comorbidity costs represent a greater burden to payers than the costs of gastric bypass surgery. Investments may be worthwhile to reduce wait times and dropout rates and improve post-surgical weight loss outcomes to save overall costs and reduce patient comorbidity prevalence.


Timely referral to the nephrologist: essential to optimizing patient outcomes.
Annual mortality on renal replacement therapy is about 10% in Western Europe and reaches 20% in the United States. The reasons responsible for this excess mortality include among others advanced age, high prevalence of diabetes and comorbid conditions, susceptibility to infections, and cancer. An additional cause that should be considered is late referral to overall renal care and for renal replacement therapy. It has been demonstrated recently that early referral may provide many advantages for the patient, such as prevention of organ damage secondary to uremia and even delay the onset of end-stage renal disease. These benefits prompted numerous recommendations for timely referral, both for dialysis and for long-term renal follow-up. Despite available guidelines for nephrology referral the current practice is still suboptimal, resulting in delayed initiation of dialysis and clinical outcomes that are not ideal. There is an urgent need in the renal community to change the current practice of referral. Beyond the benefits for patients, society may also expect potential cost effectiveness from early renal care.


Prevention of post-operative thrombosis in peripheral arteriopathies. Pentoxifylline vs. conventional antiaggregants: a six-month randomized follow-up study.
The patency rate and reocclusion incidence were studied in 97 patients following vascular surgery necessitated by occlusion in the aortoiliac or femoropopliteal regions or both. Forty-nine patients were treated orally with a combination of acetylsalicylic acid and dipyridamole (1050 mg + 150 mg/day) and 48 patients with the haemorheologically active agent pentoxifylline (1200 mg/day, Trental 400 tds.) for uniform periods of 6 months. Reocclusion occurred in 10 patients receiving ASAD and in 5 patients receiving pentoxifylline. Adverse reactions were recorded in 12 patients with ASAD (discontinuation in 11) and in 3 patients with pentoxifylline (discontinuation in 2). Patency and tolerability rate were significantly superior in those patients treated with pentoxifylline, suggesting that the combination of haemorheological effects with antihaemostaseological and antithrombotic properties offered by pentoxifylline may be of benefit in the prevention of reocclusion after vascular surgery.


Upper airway inflammation assessed by nasal lavage in compost workers: A relation with bio-aerosol exposure.
Exposure to microbial agents in the composting industry may cause work related airway inflammation. Nasal lavage (NAL) has been proposed as a noninvasive method to assess such effects in population studies. Pre- and post-shift NAL were performed in the workers of a compost plant visited in 1995 (n = 14) and 1996 (n=15), of whom only four participated in both surveys. Total cells, cytokines and other inflammation markers were measured in NAL fluid, and pre-shift levels and post/pre concentration ratios were compared with NAL results obtained in the same periods in 10 and 9 controls, respectively, and with levels of airborne exposure to microbial agents endotoxin and beta(1,3)-glucan as measured in personal air samples. Job-title specific exposure levels in the first survey ranged from 75 to 527 EU/m(3) for endotoxin and from 0.54 to 4.85 microg/m(3) for beta(1,3)-glucan. In the second survey these values were lower, 29-285 EU/m(3) and 0.36-4.44 microg/m(3), respectively. In the first survey pre-shift NAL concentrations of total cells, MPO, IL-8, NO and albumin were significantly (1.1-4.8 fold) higher in compost workers than in controls. Post/pre ratios for various markers were significantly (1.2-3.2 fold) higher in compost workers in both surveys. NAL cells were mainly neutrophils, while eosinophils were only incidentally observed. A weak relation with exposure was found for pre-shift levels of MPO, uric acid and urea in the first survey. Occupational exposure of compost workers may cause acute and possibly (sub-)chronic inflammatory reactions in the upper airways, presumably induced by non-allergenic pro-inflammatory agents like endotoxins and beta(1, 3)-glucans.


Use of cellulases and recombinant cellulose binding domains for refining TCF kraft pulp.
The modular endoglucanase Cel9B from Paenibacillus barcinonensis is a highly efficient biocatalyst, which expedites pulp refining and reduces the associated energy costs as a result. In this work, we set out to identify the specific structural domain or domains responsible for the action of this enzyme on cellulose fibre surfaces with a view to facilitating the development of new cellulases for optimum biorefining. Using the recombinant enzymes GH9-CBD3c, Fn3-CBD3b, and CBD3b, which are truncated forms of Cel9B, allowed us to assess the individual effects of the catalytic, cellulose binding, and fibronectin-like domains of the enzyme on the refining of TCF kraft pulp from Eucalyptus globulus. Based on the physico-mechanical properties obtained, the truncated form containing the catalytic domain (GH9-CBD3c) has a strong effect on fibre morphology. Comparing its effect with that of the whole cellulase (Cel9B) revealed that the truncated enzyme contributes to increasing paper strength through improved tensile strength and burst strength and also that the truncated form is more effective than the whole enzyme in improving tear resistance. Therefore, the catalytic domain of Cel9B has biorefining action on pulp. Although cellulose binding domains (CBDs) are less efficient toward pulp refining, evidence obtained in this work suggests that CBD3b alters fibre surfaces and influences paper properties as a result.


The activity of the EcoRV restriction endonuclease is influenced by flanking DNA sequences both inside and outside the DNA-protein complex.
The EcoRV restriction endonuclease cleaves DNA not only at its recognition sequence but also at most other sequences that differ from the recognition site by one base pair. Compared to the reaction at the recognition site, the reactions at noncognate sites are slow but 1 out of the 12 noncognate sites on the plasmid pAT153 is cleaved more than 50 times faster than any other. The increase in the reaction rate at the preferred noncognate site, relative to other sites, was caused by the DNA sequences in the 4 base pairs from either side of the site. For enhanced activity by EcoRV, particular bases were needed immediately adjacent to the site, inside the DNA-protein complex. At these loci, the protein interacts with the phosphate groups in the DNA and the flanking sequence may control the activity of the enzyme by determining the conformation of the DNA, thus aligning the phosphate contacts. But the preferential cleavage also depended on sequences further away from the site, at loci outside the complex. At external positions, beyond the reach of the protein, the EcoRV enzyme required flanking sequences that give rise to flexibility in DNA conformation. These may facilitate the distortion of the DNA required for catalysis by EcoRV.


Long-Term Antithrombotic Therapy and Clinical Outcomes in Patients with Acute Coronary Syndrome and Renal Impairment: Insights from EPICOR and EPICOR Asia.
Information is lacking on long-term management of patients with acute coronary syndrome (ACS) and chronic kidney disease (CKD) (estimated glomerular filtration rate [eGFR] < 60 mL/min/1.73 m2). Our objectives were to describe antithrombotic management patterns and outcomes in patients with ACS with varying renal function from the EPICOR (long-tErm follow-uP of antithrombotic management patterns In acute CORonary syndrome patients; NCT01171404) and EPICOR Asia (NCT01361386) studies. EPICOR and EPICOR Asia were prospective observational studies of patients who survived hospitalization for ACS and were enrolled at discharge in 28 countries across Europe, Latin America, and Asia. The studies were conducted from 2010 to 2013 and from 2011 to 2014, respectively. This analysis evaluated patient characteristics and oral antithrombotic management patterns and outcomes up to 2 years post-discharge according to admission eGFR: ≥ 90, 60-89, 30-59, or < 30 mL/min/1.73 m2. Among 22,380 patients with available data, eGFR < 60 mL/min/1.73 m2 was observed in 16.7%. Patients with poorer renal function were older, were at greater cardiovascular risk, and had more prior cardiovascular disease and bleeding. Patients with CKD underwent fewer cardiovascular interventions and had more in-hospital cardiovascular and bleeding events. Dual antiplatelet therapy was less likely at discharge in patients with eGFR < 30 (82.3%) than in those with ≥ 90 (91.3%) mL/min/1.73 m2 and declined more sharply during follow-up in patients with low eGFR (p < 0.0001). An adjusted proportional hazards model showed that patients with lower eGFR levels had a higher risk of cardiovascular events and bleeding. The presence of CKD in patients with ACS was associated with less aggressive cardiovascular management and an increased risk of cardiovascular events.


Biological therapy for psoriatic arthritis in clinical practice: outcomes up to 2 years.
To evaluate the performance of biological drugs in psoriatic arthritis (PsA) in a routine care setting, using the Finnish national register of biological treatment (ROB-FIN). Patients with PsA who started therapy with infliximab or etanercept between June 2000 and February 2006 (n = 127) were followed for up to 24 months. Response was evaluated using American College of Rheumatology response criteria including individual measures. Significantly diminished values for swollen and tender joints, patient's global and pain assessments, doctor's global assessment of disease activity, erythrocyte sedimentation rate, C-reactive protein, and Health Assessment Questionnaire score were observed within 3 months after commencement of both infliximab and etanercept. Values remained significantly lower throughout the 24 months of followup. ACR20 response at 3 months was 79% (n = 22/28) for infliximab and 76% (n = 34/45) for etanercept. The first biological drug was discontinued in 16% due to lack of effectiveness and in 6% due to adverse events. Anti-tumor necrosis factor-α therapy, often combined with conventional disease-modifying antirheumatic drugs, appeared to have limited toxicity and persistent effectiveness for up to 2 years in a cohort of Finnish patients with severe peripheral PsA.


Effects of an integrated medicines management program on medication appropriateness in hospitalized patients.
The effects of an integrated medicines management (IMM) program on medication appropriateness are discussed. Patients enrolled in an IMM project within a hospital were randomly allocated to either a control or an intervention group. Patients in the control group received standard pharmaceutical care, while the intervention patients received the IMM service. The appropriateness of the medications prescribed was estimated on admission and discharge using the Medicines Appropriateness Index (MAI), which ranged from 0 (no prescribing problems) to 18 (most prescribing problems). For each medication scored, the difference between admission and discharge MAI scores for individual patients was calculated. Scoring was completed on 1711 medications prescribed for 117 patients. There was a statistically significant difference (p = 0.03) in the mean MAI scores on discharge between the intervention and control groups (5.69 versus 9.97, respectively). There was also a statistically significant difference (p = 0.00003) in the mean MAI scores on admission and discharge (17.48 versus 5.69, respectively) for the intervention group. The intervention group had a mean +/- S.D. difference of -11.78 +/- 14.64, while the control group had a mean +/- S.D. difference of -3.19 +/- 11.80. The difference was statistically significant (p = 0.0011) between the two groups. This study demonstrated significant improvements in the appropriateness of medications on discharge for patients receiving an IMM service compared with patients who did not receive this service.


Multivariate proteomic analysis of the cerebrospinal fluid of patients with peripheral neuropathic pain and healthy controls - a hypothesis-generating pilot study.
Pain medicine lacks objective biomarkers to guide diagnosis and treatment. Combining two-dimensional gel proteomics with multivariate data analysis by projection, we exploratively analyzed the cerebrospinal fluid of eleven patients with severe peripheral neuropathic pain due to trauma and/or surgery refractory to conventional treatment and eleven healthy controls. Using orthogonal partial least squares discriminant analysis, we identified a panel of 36 proteins highly discriminating between the two groups. Due to a possible confounding effect of age, a new model with age as outcome variable was computed for patients (n=11), and four out of 36 protein spots were excluded due to a probable influence of age. Of the 32 remaining proteins, the following seven had the highest discriminatory power between the two groups: an isoform of angiotensinogen (upregulated in patients), two isoforms of alpha-1-antitrypsin (downregulated in patients), three isoforms of haptoglobin (upregulated in patients), and one isoform of pigment epithelium-derived factor (downregulated in patients). It has recently been hypothesized that the renin-angiotensin system may play a role in the pathophysiology of neuropathic pain, and a clinical trial of an angiotensin II receptor antagonist was recently published. It is noteworthy that when searching for neuropathic pain biomarkers with a purely explorative methodology, it was indeed a renin-angiotensin system protein that had the highest discriminatory power between patients and controls in the present study. The results from this hypothesis-generating pilot study have to be confirmed in larger, hypothesis-driven studies with age-matched controls, but the present study illustrates the fruitfulness of combining proteomics with multivariate data analysis in hypothesis-generating pain biomarker studies in humans.


Prescribing pattern of proton pump inhibitors among patients admitted to medical ward at King Abdulaziz University Hospital, Jeddah, Saudi Arabia: A retrospective study.
To determine the prescription pattern of proton pump inhibitors (PPIs) at a tertiary hospital in Saudi Arabia. This study was a retrospective cohort design using medical records at the medical ward of King Abdulaziz University Hospital (KAUH), Jeddah, Saudi Arabia. Data were collected from medical records of all patients admitted to the ward from October 2019 to March 2020. A total of 1135 medical records were retrieved, of which 709 (62.5%) were prescribed PPIs. The frequency of PPIs prescribed was highest in March 2020 (21.7%) relative to the remaining months. Omeprazole was the most prescribed PPI (84.5%), compared to pantoprazole (15.5%). The PPIs were commonly administered through oral (56.7 %), compared to intravenous route (43.3%). The PPIs were prescribed to most patients (58.4%) on the first day of admission, and 6.5% were prescribed for indications other than those approved by the Food and Drug Administration. This study demonstrates that PPIs are frequently prescribed among most patients at the medical ward of KAUH, Kingdom of Saudi Arabia. We found that the PPIs were prescribed to many patients inappropriately (not based on clinical guidelines). These results suggest the need for educational interventions among physicians on rational prescription.


[The usage of allograft nasal septal cartilage in partial laryngectomy].
Based on the experiment of five dogs, 22 cases of laryngeal carcinoma using allograft nasal septal cartilage as a framework for laryngeal defect after partial larygectomy are reported. After 5 years follow-up, the functions of phonation and respiration of 17 cases were satisfying and their phonatic loudness was raised remarkably. The method was used mainly for patients with T1 glottic carcinoma involving anterion commissure and selective T2, T3 glottic carcinoma. The advantages of allograft nasal septal cartilage in partial laryngectomy are discussed in comparison with auto nasal mucoseptal graft.


Varying regional topology within knee articular chondrocytes under simulated in vivo conditions.
Topographical cartilage variation across the knee joint has been previously reported, but there is only limited information on such gene expression profiles. Articular chondrocytes from eight different topographical regions of bovine knee joints were seeded within three-dimensional scaffolds and further cultured under static conditions (unloaded control group) or subjected to an artificial joint environment within a bioreactor (loaded group). Constructs were analyzed for glycosaminoglycan (GAG), DNA, and expression of Collagen-1,-2,-10, Aggrecan, COMP, Sox9, PRG-4, PTHrp, and MMP-1,-3,-13 mRNA after 2 weeks of in vitro culture. Exclusively among loaded constructs the overall GAG production was significantly different between regions. Patella chondrocytes had overall highest, and cells from the femoral notch had overall lowest GAG/DNA under loaded conditions. Gene expression was significantly different between regions for all targets except for Sox9, PRG-4, and PTHrp among controls and with the exception of Aggrecan, Sox9, and PTHrp among loaded samples. Under mechanical stimulation Collagen-1,-2 and Aggrecan was highest at the patello-femoral joint, whereas it was lowest at typical cartilage biopsy regions. There is a clear topographical variation among distinct regions across the knee joint for gene and matrix expression profiles under static and foremost under dynamic conditions.


NPI-2358 is a tubulin-depolymerizing agent: in-vitro evidence for activity as a tumor vascular-disrupting agent.
The diketopiperazine NPI-2358 is a synthetic analog of NPI-2350, a natural product isolated from Aspergillus sp., which depolymerizes microtubules in A549 human lung carcinoma cells. Although structurally different from the colchicine-binding site agents reported to date, NPI-2358 binds to the colchicine-binding site of tubulin. NPI-2358 has potent in-vitro anti-tumor activity against various human tumor cell lines and maintains activity against tumor cell lines with various multidrug-resistant (MDR) profiles. In addition, when evaluated in proliferating human umbilical vein endothelial cells (HUVECs), concentrations as low as 10 nmol/l NPI-2358 induced tubulin depolymerization within 30 min. Furthermore, NPI-2358 dose dependently increases HUVEC monolayer permeability--an in-vitro model of tumor vascular collapse. NPI-2358 was compared with three tubulin-depolymerizing agents with vascular-disrupting activity: colchicine, vincristine and combretastatin A-4 (CA4). Results showed that the activity of NPI-2358 in HUVECs was more potent than either colchicine or vincristine; the profile of CA4 approached that of NPI-2358. Altogether, our data show that NPI-2358 is a potent anti-tumor agent which is active in MDR tumor cell lines, and is able to rapidly induce tubulin depolymerization and monolayer permeability in HUVECs. These data warrant further evaluation of NPI-2358 as a vascular-disrupting agent in vivo. Currently, NPI-2358 is in preclinical development for the treatment of cancer.


Sexual risk behaviours and barriers to HIV testing among clients of female sex workers in Guatemala: a qualitative study.
Few interventions have targeted clients of female sex workers in Central America, despite their potential role in HIV/STI prevention. Semi-structured interviews were conducted with 30 clients of female sex workers on attitudes towards prevention of HIV/STIs, barriers to condom use and behaviour towards HIV/STI testing and treatment in Escuintla, Guatemala. Despite high knowledge of condoms as an HIV/STI preventive measure, the decision to use them was often based on the client's social judgment of the woman's sexual conduct. Regular clients reported lower condom use. Clients' risk perception diminished with the awareness of the public HIV/STI clinic addressed to female sex workers. Most preferred private clinics to increase confidentiality and were reluctant to take the HIV test for fear of a positive result. Outreach programmes offering HIV/STI counselling and testing to clients of female sex workers could increase their test uptake and health-seeking behaviour and reduce potential transmission to the general population.


Development of a novel non-radioactive cell-based method for the screening of SGLT1 and SGLT2 inhibitors using 1-NBDG.
Sodium-coupled glucose co-transporters SGLT1 and SGLT2 play important roles in intestinal absorption and renal reabsorption of glucose, respectively. Blocking SGLT2 is a novel mechanism for lowering the blood glucose level by inhibiting renal glucose reabsorption and selective SGLT2 inhibitors are under development for treatment of type 2 diabetes. Furthermore, it has been reported that perturbation of SGLT1 is associated with cardiomyopathy and cancer. Therefore, both SGLT1 and SGLT2 are potential therapeutic targets. Here we report the development of a non-radioactive cell-based method for the screening of SGLT inhibitors using COS-7 cells transiently expressing human SGLT1 (hSGLT1), CHO-K1 cells stably expressing human SGLT2 (hSGLT2), and a novel fluorescent d-glucose analogue 1-NBDG as a substrate. Our data indicate that 1-NBDG can be a good replacement for the currently used isotope-labeled SGLT substrate, (14)C-AMG. The Michaelis constant of 1-NBDG transport (0.55 mM) is similar to that of d-glucose (0.51 mM) and AMG (0.40 mM) transport through hSGLT1. The IC50 values of a SGLT inhibitor phlorizin for hSGLT1 obtained using 1-NBDG and (14)C-AMG were identical (0.11 μM) in our cell-based system. The IC50 values of dapagliflozin, a well-known selective SGLT2 inhibitor, for hSGLT2 and hSGLT1 determined using 1-NBDG were 1.86 nM and 880 nM, respectively, which are comparable to the published results obtained using (14)C-AMG. Compared to (14)C-AMG, the use of 1-NBDG is cost-effective, convenient and potentially more sensitive. Taken together, a non-radioactive system using 1-NBDG has been validated as a rapid and reliable method for the screening of SGLT1 and SGLT2 inhibitors.


Detection and determination of aromatic amines as products of reductive splitting from selected azo dyes.
The current environment-friendly regulations concerning textile products ban the marketing of textiles dyed with azo dyes capable of reductively splitting carcinogenic aromatic amines. The study analyzes seven azo dyes whose chemical structure determines various quantities of splitting aromatic amines, such as benzidine. For tests, seven commercially available azo dyes with aromatic amines in their structure were selected. These included two hazardous dyes: Acid Red 85 and Direct Blue 6, both capable of reductively splitting carcinogenic benzidine. Of the remaining five azo dyes, three--Ponceau SS, Sudan II, and Disperse Yellow 7--are capable of splitting p-phenylenediamine and aniline, while Mordant Orange 1 and Disperse Orange 3 can split only p-phenylenediamine. For Acid Red 85 and Direct Blue 6, the quantity of benzidine split from them was analyzed, depending on the conditions of the reduction process (e.g., in the HPLC method, 104 g/kg of dye for reduction in NaOH, and 41 g/kg of dye for reduction in acetate buffer). The spectrophotometric method proved useful for preliminary analysis of amine content in examined samples. Spectrophotometric analysis may be used to determine the total content of amines counted as aniline. A full qualitative and quantitative analysis of amines released from azo dyes is possible using high-performance liquid chromatography (HPLC).


Increased production of hydroxyl radical by pericentral microsomes compared to periportal microsomes after pyrazole induction of cytochrome P4502E1.
Cytochrome P4502E1 is localized in the pericentral (PC) zone of the liver acinus to a greater extent than in the periportal (PP) zone. After pyrazole treatment, PC microsomes were more active in oxidizing typical substrates of CYP2E1 than PP microsomes and had an increased content of CYP2E1. The ability of PC and PP microsomes from pyrazole-treated rats to interact with iron and generate reactive oxygen species such as the hydroxyl radical (.OH) was evaluated. A sensitive DNA strand cleavage assay was used to detect .OH; supercoiled plasmid DNA is compact but is converted by .OH-induced single strand breaks to the relaxed open circular state. Microsomes from PC hepatocytes of pyrazole-treated rats were several fold more reactive than PP microsomes in promoting NADPH-dependent DNA strand cleavage with a variety of iron catalysts, including ferric-ATP, ferric-histidine, ferric-citrate, ferric ammonium sulfate, and ferric-EDTA. DNA strand cleavage was inhibited by superoxide dismutase, catalase, and .OH scavengers such as DMSO and ethanol. Rates of H2O2 production were higher with the PC microsomes. These results indicate that rates of .OH production are higher with PC microsomes than PP microsomes after pyrazole treatment to induce cytochrome P4502E1 and suggest the possibility that elevated production of reactive oxygen species may play a role in ethanol toxicity to the PC zone of the liver acinus.


Underutilization of influenza vaccine in Jerusalem.
Influenza is an annual winter disease which causes considerable morbidity in the general population and may be lethal in defined high-risk groups. A killed vaccine, recommended for the high-risk groups as well as for several occupational categories, is moderately protective against the disease, but vaccine sales suggest that use is low. In a hospital-based convenience survey of 295 persons belonging to groups for whom influenza vaccine is recommended, 78 (26.4%) reported having received the vaccine in 1989, and 74 (25.1%) in 1990. All vaccine recipients belonged to the 244 subjects eligible because of advanced age and/or chronic illness, giving vaccination rates in this group of 32.0% in 1989 and 30.3% in 1990. None of the 51 hospital doctors and nurses interviewed had received vaccine. Among the 244 eligible because of age or illness, vaccine receipt in 1990 was related to older age and number of risk factors, but even in subjects with three separate indications for receipt of vaccine, the vaccination rate was only 48%. Among persons advised by anyone to have influenza vaccine, 62.7% received vaccine in at least one of the two years, compared to a rate of 16.4% among subjects never advised to be vaccinated. Vaccine recommendation by a medical professional increased the vaccination rate to 69.3%. Influenza vaccine uptake in vaccine eligibles in Jerusalem is low, about 30% in the sick and elderly and considerably lower in medical personnel. Since influenza vaccination can save lives, it would be worthwhile to convince the medical profession to enthusiastically recommend this vaccine to high risk individuals and to explore novel ways of increasing uptake in the high-high risk groups such as persons in hospital or those residing in institutions.


[Sun addiction, its death blow and the multitude of sunscreens].
Any mismatch between the innate cutaneous pigmentation and lifetime behaviour regarding sun exposure is a serious challenge for the skin. Sunbaths and sun parlours combine their deleterious effects. The risks for inflammatory reactions (sunburns and other photoreactions), chronic photoageing, skin field photocancerogenesis, some equivocal tumours (actinic keratosis, actinic porokeratosis,...) and overt skin cancers (basal cell carcinoma, squamous cell carcinoma, melanoma) are common conditions. For the public at large and medias, sunscreens aim at preventing these issues. Clearly, each of these goals is not fully achieved.


Deficiency in expression of the signaling protein Sin/Efs leads to T-lymphocyte activation and mucosal inflammation.
Our studies have concentrated on elucidating the role of the signaling protein Sin in T-lymphocyte function. We have previously shown that Sin overexpression inhibits T-lymphocyte development and activation. Here we show that Sin-deficient mice exhibit exaggerated immune responses characterized by enhanced cytokine secretion and T-cell-dependent antibody production. Excessive T-cell responses in young mice correlate with spontaneous development of inflammatory lesions in different organs of aged Sin(-/-) mice, particularly the small intestine. The intestinal inflammation is characterized by T- and B-cell infiltrates in the lamina propria, which correlate with crypt enlargement and marked villus expansion and/or damage. Similar to the human intestinal inflammatory disorder Crohn's disease (CD), and in contrast to most mouse models of mucosal inflammation, inflammatory lesions in the gastrointestinal tract of Sin(-/-) mice are restricted to the small bowel. Taken together, these results suggest that Sin regulates immune system and T-lymphocyte function and that immune system dysfunction in the absence of Sin may underlie the pathogenesis of tissue-specific inflammation and enteropathies such as CD.


Positive effects, side effects, and adverse events of clinical holistic medicine. A review of Gerda Boyesen's nonpharmaceutical mind-body medicine (biodynamic body-psychotherapy) at two centers in the United Kingdom and Germany.
To review adverse events of intensive, clinical holistic medicine (CHM) as it is practiced in holistic body-psychotherapy in England and Germany. Gerda Boyesen's "biodynamic body-psychotherapy" (BBP) is an intensive type of holistic mind-body medicine used by Boyesen at two centers. About 13,500 patients were treated during 1985-2005 period and studied for side effects and adverse events. The first author worked closely with Boyesen 1995-2005 with full insight in all aspects of the therapy and provided the data on side-effects. Therapy helped chronic patients with physical, psychological, sexual, psychiatric and existential problems to improve health, ability, and quality of life (NNT (number needed to treat) = 1-3). Effective in the treatment of mentally ill patients (schizophrenia, anxiety, poor mental health, low general ability). For retraumatization, brief reactive psychosis, depression, depersonalization and derealization, implanted memories, side effects from manipulations of the body, suicide/suicide attempts, hospitalization for physical and mental health problem during or 90 days after treatment, NNH (number needed to harm) > 13,500. Intensive, holistic non-drug medicine is helpful for physical, sexual, psychological, psychiatric and existential problems and is completely safe for the patient. The therapeutic value TV = NNH/NNT > 5,000. Altogether about 18,000 patients treated with different subtypes of CHM in four different countries have now been evaluated for effects, side effects and adverse events, with similar results.


Solvent Front Position Extraction with Semi-Automatic Device as a Powerful Sample Preparation Procedure Prior to Quantitative Instrumental Analysis.
The new prototype device is applied to the Solvent Front Position Extraction (SFPE) sample preparation procedure. The mobile phase is deposited onto the chromatographic plate adsorbent layer by the pipette, which is moved, according to programmed movement path, by a 3D printer mechanism. The application of the prototype device to SFPE procedure leads to the increased repeatability of the results and significant reduction of the analysis time in comparison to the classical procedure of chromatogram development. Additionally, the new equipment allows use procedures that are not possible to run using the classic chromatogram development. In this paper, the results of manual and semi-automatic sample preparation with SFPE are compared and the possible application of this prototype device is discussed.


Neurotoxic action of lead: effect on tetrahydrobiopterin metabolism in the rat.
The effects of lead, a known neurotoxin on the metabolism of a vital tetrahydrobiopterin cofactor for the hydroxylation enzymes tyrosine hydroxylase, phenylalanine hydroxylase and tryptophan hydroxylase, have been investigated. Reduced availability of this pteridine has the potential to reduce the level of the neurotransmitters noradrenaline, adrenaline and 5-hydroxytryptophan in the brain. Using the rat as a model, increases in tetrahydrobiopterin concentration and in the activity of dihydropteridine reductase, an enzyme involved in tetrahydrobiopterin metabolism, were observed after exposure to lead via the drinking water. Possible explanations for this increased level of tetrahydrobiopterin relate to alterations in the balance between synthesis and salvage of this co-factor.


Imaging in pulmonary hypertension, part 3: small vessel diseases.
Pulmonary hypertension is a significant cause of morbidity and mortality. Unfortunately, non-specific presentation and lack of awareness of the disease frequently lead to significant delay in diagnosis, often with the onset of right heart failure, when prognosis is poor and therapy is of limited effectiveness. The classification of pulmonary hypertension is a clinical one grouping diseases into categories with similar patho-physiological mechanism and therapeutic options. Pulmonary biopsy can provide a definitive diagnosis but is hazardous in patients with pulmonary hypertension. Imaging has emerged as an invaluable tool in differentiating the aetiology, assessing disease severity and directing further management. One of the most important roles of imaging is to differentiate diseases resulting from obstruction of the large pulmonary arteries from those secondary to diffuse small vessel disease, as these have very different prognosis and are also treated differently. Small vessel diseases causing pulmonary arterial hypertension most commonly result from diffuse remodelling of the pulmonary arterioles. There are multiple causes of arteriolar remodelling which share similar histopathological, clinical and imaging features. In a subgroup of small vessel diseases causing pulmonary hypertension the predominant site of increased vascular resistance is at the level of the capillaries or venules. Correct diagnosis of pulmonary veno-occlusive disease and pulmonary capillary haemangiomatosis is essential since poor prognosis and inadvertent administration of vasodilators (conventional therapy for arteriolar predominant disease) can result in fatal pulmonary oedema. Multimodality imaging plays an important role in suggesting a diagnosis, guiding further investigation and directing treatment.


Ten novel MSH2 and MLH1 germline mutations in families with HNPCC.
Hereditary nonpolyposis colorectal cancer (HNPCC) is one of the most common hereditary cancer-susceptibility syndromes. Germline mutations in mismatch repair genes are associated with the clinical phenotype of HNPCC. We report ten novel germline mutations, three in MSH2 and seven in MLH1. All but one mutation have been found in families fulfilling criteria of the Bethesda guidelines; four of them additionally fulfilled the Amsterdam criteria I or II. Eight mutations were considered pathogenic and predictive diagnostics in healthy family members at risk shall be undertaken; these include five frameshift mutations leading to premature stop codons, in MSH2: c.1672delT (p.S558Xfs) and c.2466_2467delTG (p.C822X) and in MLH1: c.1023delG (p.R341Xfs), c.1127_1128dupAT (p.K377Xfs) and c.1310delC (p.P437Xfs); three mutations leading to splice aberrations, in MSH2: c.1661G>C (r.1511_1661del) and in MLH1: c.677+3A>C (r.589_677del) and c.1990-2A>G predicted to result in a splice site defect. The remaining two mutations are unclassified variants with assumed pathogenicity: one missense mutation in the highly conserved ATPase domain of MLH1 (c.122A>G [p.D41G]) and one in-frame insertion of twelve nucleotides in MLH1 (c.2155_2156insATGTGTTCCACA [p.I719delinsNVFHI]). These two mutations were not found in 102 alleles of healthy control individuals. The corresponding tumors from all patients showed a high level of microsatellite instability (MSI-H). Immunohistochemistry (IHC) revealed complete loss of expression of the affected protein in the tumor cells from all but three patients. The tumors from the patients with the mutations c.1127_1128dupAT and c.1990-2A>G showed a reduction of expression of the MLH1-protein, rather than complete loss. In the tumor from the patient with the missense mutation c.122A>G [p.D41G] a normal expression of the proteins coded by MLH1 and MSH2 was noticed.


Increased Amygdala-Paracentral Lobule/Precuneus Functional Connectivity Associated With Patients With Mood Disorder and Suicidal Behavior.
Mood disorder patients have greater suicide risk than members of the general population, but how suicidal behavior relates to brain functions has not been fully elucidated. This study investigated how functional connectivity (FC) values between the right/left amygdala and the whole brain relate to suicidal behavior in patients with mood disorder. The participants in this study were 100 mood disorder patients with suicidal behavior (SB group), 120 mood disorder patients with non-suicidal behavior (NSB group), and 138 age- and gender-matched healthy controls (HC group). Whole-brain FC values among the three groups were compared using an analysis of covariance (ANCOVA). Compared to the NSB and HC groups, increased FC values in the right amygdala-bilateral paracentral lobule/precuneus circuit were observed in the SB group (Bonferroni-corrected, p < 0.017). The FC values in the NSB group did not differ significantly from those in the HC group (Bonferroni-corrected, p > 0.017). Moreover, there were no significant differences in FC values between mood disorder patients with suicide attempt (SA group) and mood disorder patients with suicidal ideation (SI group), while the FC values between the right amygdala and bilateral paracentral lobule/precuneus in the SA group were higher than the mean in the SI group. These findings suggest that right amygdala-paracentral lobule/precuneus dysfunction has an important role in patients with mood disorder and suicidal behavior.


Association of Aeromonas hydrophila and Vibrio alginolyticus with Larval Mortalities of Scallop (Argopecten purpuratus)
A bacteriological study was carried out in a hatchery of Argopecten purpuratus located in northern Chile which had been affected by severe larval mortalities. The phenotypic characterization of the bacterial strains revealed that Vibrio alginolyticus was the predominant species isolated in the majority of samples taken from the different units of the hatchery (microalgae, swimming larvae, seawater of larval culture tanks, and a reservoir tank of 50-μm filtered seawater). However, the bacterial population of dying larvae was composed of only Aeromonas hydrophila strains which proved to be resistant to most of the chemotherapeutic agents tested. The bioassays conducted to evaluate the effect of these bacteria on larval survival showed that all of the isolated Vibrio and Aeromonas strains possessed a high degree of pathogenicity, since they produced dying larvae on concentrations ranging from 5.5 x 10(4) to 5.5 x 10(2) cells/ml. The possible virulence mechanism of these bacteria is discussed, as well as the potential use of drugs to prevent larval mortalities.


Nerve growth factor rapidly stimulates tyrosine phosphorylation of phospholipase C-gamma 1 by a kinase activity associated with the product of the trk protooncogene.
Nerve growth factor (NGF) promotes the survival and differentiation of specific populations of neurons. The molecular mechanisms by which cells respond to NGF are poorly understood, but two clues have emerged recently. First, NGF rapidly stimulates tyrosine phosphorylation of several unidentified proteins in the NGF-responsive pheochromocytoma cell line PC12 [Maher, P. (1988) Proc. Natl. Acad. Sci. USA 85, 6788-6791]. Second, the protein-tyrosine kinase encoded by the protooncogene trk (p140trk), a member of the receptor class of tyrosine kinases, becomes activated and phosphorylated on tyrosine after NGF treatment of PC12 cells [Kaplan, D. R., Martin-Zanca, D. & Parada, L. F. (1991) Nature (London) 350, 158-160]. We now report that NGF rapidly induces tyrosine phosphorylation of phospholipase C-gamma 1 (PLC-gamma 1), and we present evidence that the responsible tyrosine kinase is either p140trk or a closely associated protein. Treatment of responsive cells with NGF elicited phosphorylation of PLC-gamma 1 on tyrosine and serine. PLC-gamma 1 immunoprecipitated from NGF-stimulated cells was phosphorylated in vitro by coprecipitating protein kinase activity, and the phosphorylations occurred principally on tyrosine. The responsible kinase could be depleted from cellular lysates by antibodies specific for p140trk. This procedure also depleted a 140-kDa protein that normally coprecipitated with PLC-gamma 1 and became phosphorylated on tyrosine in vivo in response to NGF. Analysis of tryptic peptides from PLC-gamma 1 indicated that the residues phosphorylated in vitro by p140trk-associated kinase activity were largely congruent with those phosphorylated in vivo after NGF treatment. Our findings identify PLC-gamma 1 as a likely substrate for the trk-encoded tyrosine kinase, and they provide a link between NGF-dependent activation of p140trk and the stimulation of intracellular second messenger pathways.


Direct-acting antivirals after successful treatment of early hepatocellular carcinoma improve survival in HCV-cirrhotic patients.
The effectiveness of direct-acting antivirals (DAAs) against hepatitis C virus (HCV), following successful treatment of early hepatocellular carcinoma (HCC), has been studied extensively. However, the benefit in terms of overall survival (OS) remains to be conclusively demonstrated. The aim of this study was to assess the impact of DAAs on OS, HCC recurrence, and hepatic decompensation. We prospectively enrolled 163 consecutive patients with HCV-related cirrhosis and a first diagnosis of early Barcelona Clinic Liver Cancer stage 0/A HCC, who had achieved a complete radiologic response after curative resection or ablation and were subsequently treated with DAAs. DAA-untreated patients from the ITA.LI.CA. cohort (n = 328) served as controls. After propensity score matching, outcomes of 102 DAA-treated (DAA group) and 102 DAA-untreated patients (No DAA group) were compared. In the DAA group, 7/102 patients (6.9%) died, HCC recurred in 28/102 patients (27.5%) and hepatic decompensation occurred in 6/102 patients (5.9%), after a mean follow-up of 21.4 months. OS was significantly higher in the DAA group compared to the No DAA group (hazard ratio [HR] 0.39; 95% CI0.17-0.91; p = 0.03). HCC recurrence was not significantly different between the DAA and No DAA groups (HR0.70; 95% CI0.44-1.13; p = 0.15). A significant reduction in the rate of hepatic decompensation was observed in the DAA group compared with the No DAA group (HR0.32; 95% CI0.13-0.84; p = 0.02). In the DAA group, sustained virologic response was a significant predictor of OS (HR 0.02; 95% CI 0.00-0.19; p <0.001), HCC recurrence (HR 0.25; 95% CI 0.11-0.57; p <0.001) and hepatic decompensation (HR 0.12; 95% CI 0.02-0.38; p = 0.02). In patients with HCV-related cirrhosis who had been successfully treated for early HCC, DAAs significantly improved OS compared with No DAA treatment. We aimed to determine whether direct-acting antivirals (DAAs) significantly improve overall survival in patients with hepatitis C virus-related compensated cirrhosis and a first diagnosis of hepatocellular carcinoma (HCC) which has been successfully treated with curative resection or ablation. Using propensity-score matched patients, we found that DAAs improved overall survival and reduced the risk of hepatic decompensation. However, the risk of HCC recurrence was not significantly reduced.


Maintaining Clinical Freedom Whilst Achieving Value in Biologics Prescribing: An Integrated Cross-Specialty Consensus of UK Dermatologists, Rheumatologists and Gastroenterologists.
Biologics are now key drugs in the management of immune-mediated inflammatory diseases. However, the increasingly complex biologics environment and growing cost pressures in the UK have led to variability in drug commissioning and inequity of patient access across regions. Our objectives were to provide consensus recommendations for enhancing the current situation in biologic prescribing in the UK by balancing clinical freedom with equitable distribution of biologics given the limited availability of resources. A modified Delphi approach was used to reach integrated, cross-specialty consensus among dermatologists, rheumatologists and gastroenterologists practising within the English National Health Service (NHS). We describe the concepts of clinical freedom and clinical judgement and demonstrate how, together with patient choice, they can be exercised in the context of biologic prescribing in the NHS. We highlight that in England, local variations occur that are at odds with National Institute for Health and Care Excellence (NICE) guidance; these variably limit the degree to which clinicians can exercise clinical freedom and impact on equity of patient access to treatments. We define factors encompassing a drug's value and identify challenges to the measurement and interpretation of this concept, which can raise barriers to the freedom of clinical choice and appropriate prescribing decisions allowing practices of holistic and personalised medicine. Cross-specialty consensus recommendations on ensuring equitable access to biologics in the NHS while protecting appropriate and individualised drug selection for patients are provided. We have also provided strategies for improving physician-commissioner communication to harmonise equity of patient access to biologics across England and improve patient outcomes. Commentary from patient advisory groups indicates that they welcome our exploration that value does not equal cost and agree that there should be an emphasis on shared decision making, which requires the clinician to practice clinical freedom by aligning the patient's needs and preferences with available treatment choices. This consensus highlights the need to strike a balance between clinical freedom and short-term cost restrictions to support equitable resource distribution within the English NHS. Consideration of these recommendations may help to harmonise local, regional and national services and balance equity of patient access to biologic treatments with excellence in the NHS.


TREM-1 multimerization is essential for its activation on monocytes and neutrophils.
The triggering receptor expressed on myeloid cells-1 (TREM-1) is a receptor expressed on innate immune cells. By promoting the amplification of inflammatory signals that are initially triggered by Toll-like receptors (TLRs), TREM-1 has been characterized as a major player in the pathophysiology of acute and chronic inflammatory diseases, such as septic shock, myocardial infarction, atherosclerosis, and inflammatory bowel diseases. However, the molecular events leading to the activation of TREM-1 in innate immune cells remain unknown. Here, we show that TREM-1 is activated by multimerization and that the levels of intracellular Ca2+ release, reactive oxygen species, and cytokine production correlate with the degree of TREM-1 aggregation. TREM-1 activation on primary human monocytes by LPS required a two-step process consisting of upregulation followed by clustering of TREM-1 at the cell surface, in contrast to primary human neutrophils, where LPS induced a rapid cell membrane reorganization of TREM-1, which confirmed that TREM-1 is regulated differently in primary human neutrophils and monocytes. In addition, we show that the ectodomain of TREM-1 is able to homooligomerize in a concentration-dependent manner, which suggests that the clustering of TREM-1 on the membrane promotes its oligomerization. We further show that the adapter protein DAP12 stabilizes TREM-1 surface expression and multimerization. TREM-1 multimerization at the cell surface is also mediated by its endogenous ligand, a conclusion supported by the ability of the TREM-1 inhibitor LR12 to limit TREM-1 multimerization. These results provide evidence for ligand-induced, receptor-mediated dimerization of TREM-1. Collectively, our findings uncover the mechanisms necessary for TREM-1 activation in monocytes and neutrophils.


Sensitization to the lysosomal cell death pathway by oncogene-induced down-regulation of lysosome-associated membrane proteins 1 and 2.
Expression and activity of lysosomal cysteine cathepsins correlate with the metastatic capacity and aggressiveness of tumors. Here, we show that transformation of murine embryonic fibroblasts with v-H-ras or c-src(Y527F) changes the distribution, density, and ultrastructure of the lysosomes, decreases the levels of lysosome-associated membrane proteins (LAMP-1 and LAMP-2) in an extracellular signal-regulated kinase (ERK)- and cathepsin-dependent manner, and sensitizes the cells to lysosomal cell death pathways induced by various anticancer drugs (i.e., cisplatin, etoposide, doxorubicin, and siramesine). Importantly, K-ras and erbb2 elicit a similar ERK-mediated activation of cysteine cathepsins, cathepsin-dependent down-regulation of LAMPs, and increased drug sensitivity in human colon and breast carcinoma cells, respectively. Notably, reconstitution of LAMP levels by ectopic expression or by cathepsin inhibitors protects transformed cells against the lysosomal cell death pathway. Furthermore, knockdown of either lamp1 or lamp2 is sufficient to sensitize the cells to siramesine-induced cell death and photo-oxidation-induced lysosomal destabilization. Thus, the transformation-associated ERK-mediated up-regulation of cysteine cathepsin expression and activity leads to a decrease in the levels of LAMPs, which in turn contributes to the enhanced sensitivity of transformed cells to drugs that trigger lysosomal membrane permeabilization. These data indicate that aggressive cancers with high cysteine cathepsin levels are especially sensitive to lysosomal cell death pathways and encourage the further development of lysosome-targeting compounds for cancer therapy.


Modelling Zika Virus Infection of the Developing Human Brain In Vitro Using Stem Cell Derived Cerebral Organoids.
The recent emergence of Zika virus (ZIKV) in susceptible populations has led to an abrupt increase in microcephaly and other neurodevelopmental conditions in newborn infants. While mosquitos are the main route of viral transmission, it has also been shown to spread via sexual contact and vertical mother-to-fetus transmission. In this latter case of transmission, due to the unique viral tropism of ZIKV, the virus is believed to predominantly target the neural progenitor cells (NPCs) of the developing brain. Here a method for modeling ZIKV infection, and the resulting microcephaly, that occur when human cerebral organoids are exposed to live ZIKV is described. The organoids display high levels of virus within their neural progenitor population, and exhibit severe cell death and microcephaly over time. This three-dimensional cerebral organoid model allows researchers to conduct species-matched experiments to observe and potentially intervene with ZIKV infection of the developing human brain. The model provides improved relevance over standard two-dimensional methods, and contains human-specific cellular architecture and protein expression that are not possible in animal models.


High-efficiency genome editing using a dmc1 promoter-controlled CRISPR/Cas9 system in maize.
Previous studies revealed that the promoters for driving both Cas9 and sgRNAs are quite important for efficient genome editing by CRISPR/Cas9 in plants. Here, we report our results of targeted genome editing using the maize dmc1 gene promoter combined with the U3 promoter for Cas9 and sgRNA, respectively. Three loci in the maize genome were selected for targeting. The T0 plants regenerated were highly efficiently edited at the target sites with homozygous or bi-allelic mutants accounting for about 66%. The mutations in T0 plants could be stably transmitted to the T1 generation, and new mutations could be generated in gametes or zygotes. Whole-genome resequencing indicated that no off-target mutations could be detected in the predicted loci with sequence similarity to the targeted site. Our results show that the dmc1 promoter-controlled (DPC) CRISPR/Cas9 system is highly efficient in maize and provide further evidence that the optimization of the promoters used for the CRISPR/Cas9 system is important for enhancing the efficiency of targeted genome editing in plants. The evolutionary conservation of the dmc1 gene suggests its potential for use in other plant species.