Hormetic effect of methylmercury on Caenorhabditis elegans.
Research has demonstrated the toxic effects of methylmercury (MeHg), yet molecular mechanisms underlying its toxicity are not completely understood. Caenorhabditis elegans (C. elegans) offers a unique biological model to explore mechanisms of MeHg toxicity given many advantages associated with its ease of use and genetic power. Since our previous work indicated neurotoxic resistance of C. elegans to MeHg, the present study was designed to examine molecular mechanisms associated with this resistance. We hypothesized MeHg would induce expression of gst, hsp or mtl in vivo since glutathione (GSH), heat shock proteins (HSPs), and metallothioneins (MTs) have shown involvement in MeHg toxicity. Our studies demonstrated a modest, but significant increase in fluorescence in gst-4::GFP and mtl-1::GFP strains at an acute, low L1 MeHg exposure, whereas chronic L4 MeHg exposure induced expression of gst-4::GFP and hsp-4::GFP. Knockout gst-4 animals showed no alterations in lethality sensitivity compared to wildtype animals whereas mtl knockouts displayed increased sensitivity to MeHg exposure. GSH levels were increased by acute MeHg treatment and depleted with chronic exposure. We also demonstrate that MeHg induces hormesis, a phenotype whereby a sublethal exposure to MeHg rendered C. elegans resistant to subsequent exposure to the organometal. The involvement of gst-4, hsp-4, mtl-1, and mtl-2 in hormesis was examined. An increase in gst-4::GFP expression after a low-dose acute exposure to MeHg indicated that gst-4 may be involved in this response. Our results implicate GSH, HSPs, and MTs in protecting C. elegans from MeHg toxicity and show a potential role of gst-4 in MeHg-induced hormesis.


Early Experience and Lessons Learned Using Implanted Hemodynamic Monitoring in Patients With Fontan Circulation.
Data on the use of implanted hemodynamic monitoring (IHM) in patients with Fontan circulation are limited. This study reports our experience using the CardioMEMS HF system in adults with Fontan circulation. This single-center, retrospective study evaluated heart failure hospitalizations, procedural complications, and device-related complications in patients with Fontan circulation referred for IHM placement (2015-2022). The association of pulmonary artery pressure (by most recent catheterization and median IHM pressure within 30 days of placement) with both death and follow-up Model for End-Stage Liver Disease Excluding International Normalized Ratio score were evaluated. Of 18 patients referred for IHM placement, 17 were successful (median age, 30 [range 21-48] years, 6 women). Procedural complications (access site hematomas, pulmonary artery staining) occurred in 3 patients, without device-related procedural complications. In follow-up (median, 35 [range, 6-83] months), 1 patient developed a pulmonary embolism (possibly device-related). Heart failure hospitalizations/year were similar before and after IHM (median, 1 [interquartile range, 0-1.0] versus 0.6 [0-2.3]; P=0.268), though only 46% of heart failure hospitalizations had associated IHM transmissions. IHM pressures were associated with Model for End-Stage Liver Disease Excluding International Normalized Ratio scores (R2=0.588, P<0.001), though catheterization pressures were not (R2=0.140, P=0.139). The long-term mortality rate was 53% in this cohort. On unadjusted survival analysis, IHM pressures ≥18 mm Hg were associated with mortality (log rank P=0.041), which was not reproduced with catheterization pressures (log rank P=0.764). In patients with Fontan circulation, IHM did not reduce heart failure hospitalizations, though patient adherence to transmission was low. Device-related complications were low. IHM pressures may better represent real-life conditions compared with catheterization given associations with mortality and Model for End-Stage Liver Disease Excluding International Normalized Ratio score.


[Reversion of oral anticoagulation in emergency surgery].
Bleeding is the most serious adverse effect of oral anticoagulants. Emergency surgery requires the rapid reversion of coagulation disorder though the use of such products as prothrombin complex, vitamin K, or activated recombinant factor VII. We analyze the cases of 5 patients on oral anticoagulants (vitamin K antagonists) who required emergency surgery in our hospital in the past year. We describe patient characteristics, laboratory findings before and after correction of anticoagulant effects, preoperative treatment of coagulation disorder, surgical interventions, and outcomes.


Distinct epigenomic patterns are associated with haploinsufficiency and predict risk genes of developmental disorders.
Haploinsufficiency is a major mechanism of genetic risk in developmental disorders. Accurate prediction of haploinsufficient genes is essential for prioritizing and interpreting deleterious variants in genetic studies. Current methods based on mutation intolerance in population data suffer from inadequate power for genes with short transcripts. Here we show haploinsufficiency is strongly associated with epigenomic patterns, and develop a computational method (Episcore) to predict haploinsufficiency leveraging epigenomic data from a broad range of tissue and cell types by machine learning methods. Based on data from recent exome sequencing studies on developmental disorders, Episcore achieves better performance in prioritizing likely-gene-disrupting (LGD) de novo variants than current methods. We further show that Episcore is less-biased by gene size, and complementary to mutation intolerance metrics for prioritizing LGD variants. Our approach enables new applications of epigenomic data and facilitates discovery and interpretation of novel risk variants implicated in developmental disorders.


Haptics - touchfeedback technology widening the horizon of medicine.
Haptics, or touchsense haptic technology is a major breakthrough in medical and dental interventions. Haptic perception is the process of recognizing objects through touch. Haptic sensations are created by actuators or motors which generate vibrations to the users and are controlled by embedded software which is integrated into the device. It takes the advantage of a combination of somatosensory pattern of skin and proprioception of hand position. Anatomical and diagnostic knowledge, when it is combined with this touch sense technology, has revolutionized medical education. This amalgamation of the worlds of diagnosis and surgical intervention adds precise robotic touch to the skill of the surgeon. A systematic literature review was done by using MEDLINE, GOOGLE SEARCH AND PubMed. The aim of this article was to introduce the fundamentals of haptic technology, its current applications in medical training and robotic surgeries, limitations of haptics and future aspects of haptics in medicine.


The experience of nutritional care according to patients with head and neck cancer involved with a combined dietitian, specialist nurse and speech pathologist clinic in a regional Australia: a qualitative longitudinal study.
Malnutrition is a co-morbidity of head and neck cancer (HNC) that has negative consequences for patients. Evidence-based guidelines (EBGs) provide recommendations to prevent and manage malnutrition. A clinic that combines the services of a dietitian, specialist oncology nurse and speech pathologist may promote the implementation of nutritional EBGs in regional Australia. This study aimed to explore the nutritional care experience that patients with HNC had in this setting. A qualitative longitudinal study collected data via semi-structured interviews with HNC patients who were treated in one regional cancer care network in Australia. Interviews were conducted at key points in their HNC journey from diagnosis to 4 months post-radiotherapy. Data was analysed using a grounded theory approach. Ten participants completed a total of thirty-six interviews. The findings were grouped into four categories: "preparing for nutritional challenges", "multidisciplinary care directed by patient needs", "the battle to eat", and "incongruence between patient values and nutritional priorities". These findings highlight the nutritional burden associated with HNC and barriers to patients accepting nutritional support from healthcare professionals. Information provided by doctors and nurses prior to treatment may help patients prepare for the nutritional challenges ahead and accept support from dietitians. Furthermore, clinics that promote continuity through treatment and allow dietitians to lead aspects of nutritional care, in collaboration with nurses, speech pathologists and doctors, may also enhance the nutritional care experience. More qualitative research within HNC teams would provide further insight on enhancing the implementation of nutritional EBGs to improve outcomes for these patients.


Regional correlation of PET and CT in senile dementia of the Alzheimer type.
Alzheimer disease is manifested by both widespread and regionally restricted brain changes, some of which have recently been identified in vivo with computed tomography (CT) and positron emission tomography (PET). This is a report of the regional correlation of CT and PET measurements in 19 carefully diagnosed subjects comprising 11 controls and eight patients with senile dementia of the Alzheimer type. Regional CT attenuation values did not discriminate between the two groups, but PET using 18F-2-deoxy-2-fluoro-D-glucose demonstrated significant regional reductions (range, 21%-28%) in glucose utilization in the Alzheimer group. PET measures were also more consistently related to cognitive decline. The correlation between CT structural measures and PET metabolic measures demonstrated consistent relations between widespread PET regions and CT changes in the thalamus, posterior limb of the internal capsule, and temporal lobes. However, CT changes in the frontal white matter, caudate nucleus, and anterior limb of the internal capsule were not related to any regional PET changes. These data support previous findings of temporal lobe involvement in Alzheimer disease and suggest the involvement of structures in the region of the third ventricle.


Comparative sequence analysis of the long repeat regions and adjoining parts of the long unique regions in the genomes of herpes simplex viruses types 1 and 2.
We report the determination of the DNA sequence of the long repeat (RL) region and adjacent parts of the long unique (UL) region in the genome of herpes simplex virus type 2 (HSV-2) strain HG52. The DNA sequences and genetic content of the extremities of HSV-2 UL were found to be closely similar to those determined previously for HSV-1. The 5658 bp sequenced at the left end of HSV-2 UL contained coding regions for genes UL1 to UL4 plus part of UL5. The 4355 bp sequenced at the right end of UL contained coding regions for part of gene UL53, and the whole of genes UL54 to UL56. Comparison of the HSV-1 and HSV-2 UL56 sequences led to a correction in the published HSV-1 UL56 reading frame. The HSV-2 RL region, including one copy of the a sequence, was determined to be 9263 bp, with a base composition of 75.4% G+C and with many repetitive sequence elements. In HSV-2 RL, sequences were identified corresponding to HSV-1 genes encoding the immediate early IE110 (ICP0) transcriptional regulator and the ICP34.5 neurovirulence factor; the former HSV-2 gene was proposed to contain two introns, and the latter one intron. Downstream of the HSV-2 immediate early gene, the RL sequence encoding the latency-associated transcripts (LATs) was found to be dissimilar to that in HSV-1; the probable LAT promoter regions, however, showed similarities to HSV-1. Properties of the LAT sequences in both HSV-1 and HSV-2 were consistent with LATs being generated as an intron excised from a longer transcript.


Asian pollution climatically modulates mid-latitude cyclones following hierarchical modelling and observational analysis.
Increasing levels of anthropogenic aerosols in Asia have raised considerable concern regarding its potential impact on the global atmosphere, but the magnitude of the associated climate forcing remains to be quantified. Here, using a novel hierarchical modelling approach and observational analysis, we demonstrate modulated mid-latitude cyclones by Asian pollution over the past three decades. Regional and seasonal simulations using a cloud-resolving model show that Asian pollution invigorates winter cyclones over the northwest Pacific, increasing precipitation by 7% and net cloud radiative forcing by 1.0 W m(-2) at the top of the atmosphere and by 1.7 W m(-2) at the Earth's surface. A global climate model incorporating the diabatic heating anomalies from Asian pollution produces a 9% enhanced transient eddy meridional heat flux and reconciles a decadal variation of mid-latitude cyclones derived from the Reanalysis data. Our results unambiguously reveal a large impact of the Asian pollutant outflows on the global general circulation and climate.


The effect of increases in the Syrian refugee population in Turkey on public maternal and child health outcomes.
Our purpose in this study is to explore the effect that increases in the Syrian refugee population could have on public maternal and child health outcomes. Data collected from official records from 81 Turkish provinces in 2018, and then, subjected to mediating analysis using a path analytic approach to examine the interrelationships between the Syrian population, adolescent marriage, and economic integration factors on refugee maternal and child health outcomes. An increase in the Syrian population would lead to a rise in adolescent birth rates and under-5 mortality under the mediating effect of an increase in adolescent female marriage.


An innovative nanoformulation utilizing tumor microenvironment-responsive PEG-polyglutamic coating and dynamic charge adjustment for specific targeting of ER stress inducer, microRNA, and immunoadjuvant in pancreatic cancer: In vitro investigations.
Pancreatic ductal adenocarcinoma (PDAC) is a significant obstacle to lowering global cancer deaths. CB-5083, a novel valosin-containing protein (VCP)/p97 inhibitor that disrupts proteasomal degradation and induces endoplasmic reticulum stress (ERS) accumulation, was evaluated as an inducer of immunogenic cell death (ICD) in PDAC treatment. Furthermore, miR-142 enhances checkpoint blockade and promotes M1 repolarization, while Toll-like receptor 7/8 agonist resiquimod (R) acts as an immunoadjuvant to amplify the immune response to miR-142. This research signifies the first integration of CB, miR-142, and R in solid lipid nanoparticles (SLNs) modified with peptides targeting PD-L1, EGFR, and ER, which were shelled by the PEG-polyglutamic (PGA) coating that detaches in response to the acidic pH values in the tumor microenvironment (TME). The modified SLNs exhibited pH-sensitive cytotoxicity against Panc-02 cells, preserving normal cells and preventing hemolysis. The innovative approach simultaneously modulated pathways, including VCP/Bip/K48-Ub/ATF6, IRE1α/XBPs/LC3II, PD-L1/TGF-β/IL-10/CD206/MSR1/Arg1, and TNF-α/IFN-γ/IL-6/iNOS/COX-2. Combined treatment blocked VCP, arrested the cell cycle, inhibited EMT, triggered ERS-mediated autophagy/apoptosis, and stimulated robust ICD via the release of damage-associated molecular patterns. This adaptable nanoformulation, displaying pH-sensitive PEG-PGA de-coating and precisely targeting EGFR, PD-L1, and ER, serves to hinder EMT and immune evasion, subsequently amplifying ICD in PDAC cells and the TME.


Protective Headgear Attenuates Forces on the Inner Table and Pressure in the Brain Parenchyma During Blast and Impact: An Experimental Study Using a Simulant-Based Surrogate Model of the Human Head.
Military personnel sustain head and brain injuries as a result of ballistic, blast, and blunt impact threats. Combat helmets are meant to protect the heads of these personnel during injury events. Studies show peak kinematics and kinetics are attenuated using protective headgear during impacts; however, there is limited experimental biomechanical literature that examines whether or not helmets mitigate peak mechanics delivered to the head and brain during blast. While the mechanical links between blast and brain injury are not universally agreed upon, one hypothesis is that blast energy can be transmitted through the head and into the brain. These transmissions can lead to rapid skull flexure and elevated pressures in the cranial vault, and, therefore, may be relevant in determining injury likelihood. Therefore, it could be argued that assessing a helmet for the ability to mitigate mechanics may be an appropriate paradigm for assessing the potential protective benefits of helmets against blast. In this work, we use a surrogate model of the head and brain to assess whether or not helmets and eye protection can alter mechanical measures during both head-level face-on blast and high forehead blunt impact events. Measurements near the forehead suggest head protection can attenuate brain parenchyma pressures by as much as 49% during blast and 52% during impact, and forces on the inner table of the skull by as much as 80% during blast and 84% during impact, relative to an unprotected head.


[Manifestations of colonic dysbacteriosis in patients with drug-resistant respiratory tuberculosis].
The bacteriological and clinical manifestations of colonic dysbacteriosis were studied in 65 patients with drug-resistant respiratory tuberculosis before treatment with reserve drugs. Colonic microbiological disturbances corresponding to first-, second-, and third-degree dysbacteriosis were detected in 6 (9%), 14 (21%), and 42 (65%), respectively; no evidence of dysbacteriosis was found in 3 (5%) patients. There were characteristic clinical symptoms of dysbacteriosis (tympanism in 80% of the patients, signs of dyspepsia in 73.8%, sensation of abdominal discomfort in 69.2%, stool pattern changes in 65%). The magnitude of dysbiotic disorders was found to be associated with the severity of the tuberculous process (the extent and presence of lung tissue destruction) and the duration of previous standard chemotherapy. Intoxication and severe colonic microenvironmental disturbances were ascertained to have a mutual impact. Gastrointestinal comorbidity in remission did not affect the severity of colonic dysbacteriosis in patients with drug-resistant pulmonary tuberculosis.


[Inhibition of thrombocyte aggregation after heart valve replacement].
After implantation of a mechanical cardiac valve, lifelong, full anticoagulation has been the tradition. After implantation of a biological valve, anticoagulation during 3 months subsequent to the operation is customary. This review evaluates the role of platelet inhibition after cardiac valve replacement. Platelet aggregation is inhibited effectively by aspirin in a daily dose of 100-160 mg. At this dose, episodes of severe bleeding are not significantly more frequent than during placebo, whereas patients on full anticoagulation bleed at a rate of 2% per year. After implantation of a mechanical cardiac valve, sole platelet inhibition is inferior to full anticoagulation. With a lower rate of bleeding, aspirin appears to prevent thromboembolic episodes caused by mechanical bileaflet prostheses (e.g., St. Jude) in the aortic position, and is almost as efficient as full anticoagulation. In Germany, patients with a porcine bioprosthesis, as currently in use, in the aortic position, frequently receive 100 mg aspiring per day. For patients with porcine bioprostheses in the mitral plus eventually in the aortic position in stable sinus rhythm, 100 mg aspirin per day is preferred to anticoagulation. For children with mechanical aortic valves, aspirin (2 mg/kg/day) needs to be considered an effective and convenient alternative to anticoagulation. Combining anticoagulation with 100 mg aspirin per day after implantation of a left-sided mechanical cardiac valve is pathophysiologically sound, but used to be considered as rendering the patients too bleeding-prone. Recently, total mortality and morbidity definitely have been demonstrated to be reduced by combined anticoagulation and platelet inhibition as compared to sole anticoagulation. Combining platelet inhibition with moderate anticoagulation (INR 2.0-3.0) was superior to combining it with full anticoagulation in terms of safety (INR > 3.0). Thus, current evidence favors combining moderate anticoagulation with 100 mg aspirin per day after left-sided mechanical cardiac valve replacement.


Interventions to support people with dementia and their caregivers during the transition from home care to nursing home care: A systematic review.
During the transition of people with dementia from home to nursing home family caregivers often feel burdened. We aimed to 1) identify interventions which support people with dementia and their caregivers in the transition from home care to nursing home care, 2) synthesize the evidence for efficacy of these interventions, and 3) examine whether the identified interventions have been systematically developed, evaluated and implemented according to the Medical Research Council guidance on complex interventions. A systematic review of randomised controlled trials was conducted according to the recommendations specified in the Cochrane Handbook for Intervention Reviews. The review protocol was registered in PROSPERO (2015: CRD42015019839). Reporting follows the Preferred Reporting Items for Systematic Reviews and Meta-Analyses: the PRISMA statement. MEDLINE, CENTRAL, PsycINFO, CINAHL, OTseeker, and PEDro, were searched. Other sources included Google Scholar, and ALOIS. Two reviewers independently assessed the eligibility of the articles. Data extraction was performed by one reviewer and verified independently by another. The Cochrane Risk of Bias tool was used for critical appraisal. Development and evaluation of the identified interventions were assessed, taking the Medical Research Council guidance into account. Review findings were synthesized narratively. The search yielded 1278 records. Five studies were included, all conducted in the United States (4 RCTs and 1 cRCT with a total of 695 participants). The psychosocial interventions were individual and family counseling via telephone or ad hoc all of which addressed only informal caregivers. The intervention components, content and mode of delivery differed widely with inconsistent results. Significant intervention effects were found for the reduction of caregivers' depressive symptoms, burden, feeling of guilt, emotional distress, overload, and interactions with staff. Other outcomes, i.e. stress, placement adaptation, role overload, and role captivity, were not statistically significantly affected. The assessment for bias risk across studies varied from moderate to low. Only two studies tested the feasibility of the intervention before full scale evaluation, none evaluated the implementation process according to the Medical Research Council framework. We identified only a few studies with heterogeneous outcomes; evidence regarding the effectiveness of psychosocial interventions is thus insufficient. Further research is needed focusing on the development and evaluation of complex psychosocial interventions and more well-designed RCTs with larger sample sizes based on a rigorous methodology. Reporting on feasibility and implementation processes of interventions should be guaranteed, since it is crucial to evaluate transferability across care settings.


Ultrafast laser-pulse transmission and imaging through biological tissues.
The transmission of 100-fs ultrafast laser pulses through biological tissues was measured by using femtosecond and picosecond time-resolved detection techniques. The broadening of transmitted pulses was found to increase as the thickness of the biological tissue increases. The absence of a distinct ballistic pulse transmitted through a relatively thin tissue is in sharp contrast with the pulse transmission through a random medium of discrete scatterers. Because of the continuous variation of the dielectric constant in tissue, the photons undergo scattering through the tissue, travel in various small zigzag least optical paths, and form a broadened early-arriving portion of the transmitted pulse. Even in the absence of a well-defined ballistic pulse, we can image an opaque object hidden inside a tissue as thick as 6.5 mm with submillimeter resolution by selecting the early-arriving portion of the transmitted pulse.


Hemophilus influenzae pneumonia in adults: report of five cases caused by ampicillin-resistant strains.
Recently there has been increased recognition of Hemophilus influenzae as a cause of pneumonia in adults. Although ampicillin-resistant strains of Hemophilus influenzae have been a major problem in pediatric practice, such strains have not previously been noted to be a significant problem in the treatment of adult pneumonia. We report 5 cases of pneumonia caused by beta-lactamase-producing strains of Hemophilus influenzae. These organisms were susceptible to chloramphenicol but resistant to ampicillin. Cure was achieved by treatment with chloramphenicol after the initial treatment with ampicillin had failed. The ability of a microbiology laboratory to isolate and to test routinely for ampicillin-resistant strains is an important factor in the successful treatment of Hemophilus influenzae infections.


A new mathematical approach to finding global solutions of the magnetic structure determination problem.
Determination of magnetic structure is an important analytical procedure utilized in various fields ranging from fundamental condensed-matter physics and chemistry to advanced manufacturing. It is typically performed using a neutron diffraction technique; however, finding global solutions of the magnetic structure optimization problem represents a significant challenge. Generally, it is not possible to mathematically prove that the obtained magnetic structure is a truly global solution and that no solution exists when no acceptable structure is found. In this study, the global optimization technique called semidefinite relaxation of quadratic optimization, which has attracted much interest in the field of applied mathematics, is proposed to use as a new analytical method for the determination of magnetic structure, followed by the application of polarized neutron diffraction data. This mathematical approach allows avoiding spurious local solutions, decreasing the amount of time required to find a tentative solution and finding multiple solutions when they exist.


Hydrogen-Borrowing Alcohol Bioamination with Coimmobilized Dehydrogenases.
The amination of alcohols is an important transformation in chemistry. The redox-neutral (i.e., hydrogen-borrowing) asymmetric amination of alcohols is enabled by the combination of an alcohol dehydrogenase (ADH) with an amine dehydrogenase (AmDH). In this work, we enhanced the efficiency of hydrogen-borrowing biocatalytic amination by co-immobilizing both dehydrogenases on controlled porosity glass FeIII ion-affinity beads. The recyclability of the dual-enzyme system was demonstrated (5 cycles) with total turnover numbers of >4000 and >1000 for ADH and AmDH, respectively. A set of (S)-configured alcohol substrates was aminated with up to 95% conversion and >99%ee (R). Preparative-scale amination of (S)-phenylpropan-2-ol resulted in 90% conversion and 80% yield of the product in 24 h.


Detection of enzymatic activities in sodium dodecyl sulfate-polyacrylamide gels: DNA polymerases as model enzymes.
Recent techniques for detecting the catalytic activity of enzymes in sodium dodecyl sulfate (SDS)-polyacrylamide gels have been hampered by lack of reproducibility associated with variability in commercial SDS preparations. Simple expedients which facilitate reproducible detection of DNA polymerase activity and which appear to be widely applicable to detection of other enzymes are reported here. It was observed that reproducibility of a reported procedure for DNA polymerase detection (Spanos, A., Sedgwick, S. G., Yarranton, G. T., Hübscher, U., and Banks, G. R. (1981) Nucl. Acids Res. 9, 1825-1839) depends on the SDS used for electrophoresis, and that sensitivity is markedly reduced if currently available SDS is substituted for the discontinued product specified by Spanos et al. A modified procedure yielding sensitivity with contemporary commercial SDS, which exceeds the sensitivity observed when using the protocol and the SDS originally specified, is described. The modifications employed, which presumably promote renaturation of enzymes, are (1) embedding fibrinogen in gels and (2) washing detergent from gels with aqueous isopropanol after electrophoresis. These expedients permit detection of picogram amounts of Escherichia coli DNA polymerase 1 and its Klenow fragment and nanogram amounts of calf thymus alpha and rat liver (Novikoff hepatoma) beta polymerases. Finally, it is shown that sensitivity of DNA polymerase detection is reduced by lipophilic contaminants in contemporary commercial SDS, and that the expedients employed here mitigate the deleterious effect of these impurities.


The ITAG Spinal Simulator (ISS): A New Simulator for Difficult Lumbar Punctures.
Lumbar punctures present a specific challenge in various medical specialties; appropriate simulators need to take into account the specific technical difficulties related to a realistic patient population, but currently fail to address the variety of anatomical differences seen in practice. We interviewed several leaders in the field of anesthesiology with extensive experience in lumbar puncture procedures, subsequently developing a more realistic training simulator. This novel simulator was built using silicone-based materials and advanced 3D-printing techniques, specifically tailored to be capable of mimicking a variety of patient populations without having to dispose of essential components after each use. Two Anesthesiologists with at least 20 years of experience were asked to perform several spinal tap procedures. Following testing, experts rated the simulator based on its procedural realism, usefulness in improving skill set, and overall simulation efficacy.The gathered validation outcomes based on the questionnaire evaluations completed by experts show excellent preliminary results, with an overall mean score of 4.8 out of 5 (96%). These preliminary results highlight the potential for the simulator's application as a tool to improve medical simulation education and future patient outcomes.


Chemoradiotherapy Increases Intratumor Heterogeneity of HPSE Expression in the Relapsed Glioblastoma Tumors.
Adjuvant chemoradiotherapy is a standard treatment option for glioblastoma multiforme (GBM). Despite intensive care, recurrent tumors developed during the first year are fatal for the patients. Possibly contributing to this effect, among other causes, is that therapy induces changes of polysaccharide heparan sulfate (HS) chains in the cancer cells and/or tumor microenvironment. The aim of this study was to perform a comparative analysis of heparanase (HPSE) expression and HS content in different normal and GBM brain tissues. Immunohistochemical analysis revealed a significant decrease of HPSE protein content in the tumor (12-15-fold) and paratumorous (2.5-3-fold) GBM tissues compared with normal brain tissue, both in cellular and extracellular compartments. The relapsed GBM tumors demonstrated significantly higher intertumor and/or intratumor heterogeneity of HPSE and HS content and distribution compared with the matched primary ones (from the same patient) (n = 8), although overall expression levels did not show significant differences, suggesting local deterioration of HPSE expression with reference to the control system or by the treatment. Double immunofluorescence staining of various glioblastoma cell lines (U87, U343, LN18, LN71, T406) demonstrated a complex pattern of HPSE expression and HS content with a tendency towards a negative association of these parameters. Taken together, the results demonstrate the increase of intratumor heterogeneity of HPSE protein in relapsed GBM tumors and suggest misbalance of HPSE expression regulation by the adjuvant anti-GBM chemoradiotherapy.


United States birth weight reference corrected for implausible gestational age estimates.
To provide an updated US birth weight for gestational age reference corrected for likely errors in last menstrual period (LMP)-based gestational age dating, as well as means and SDs, to enable calculation of continuous and categorical measures of birth weight for gestational age. From the 2009-2010 US live birth files, we abstracted singleton births between 22 and 44 weeks of gestation with at least 1 nonmissing estimate of gestational age (ie, LMP or obstetric/clinical) and birth weight. Using an algorithm based on birth weight and the concordance between these gestational age estimates, implausible LMP-based gestational age estimates were either excluded or corrected by using the obstetric/clinical estimate. Gestational age- and sex-specific birth weight means, SDs, and smoothed percentiles (3rd, 5th, 10th, 90th, 95th, 97th) were calculated, and the 10th and 90th percentiles were compared with published population-based references. A total of 7 818 201 (99% of eligible) births were included. The LMP-based estimate of gestational age comprised 85% of the dataset, and the obstetric/clinical estimate comprised the remaining 15%. Cut points derived from the current reference identified ∼10% of births as ≤10th and ≥90th percentiles at all gestational weeks, whereas cut points derived from previous US-based references captured variable proportions of infants at these thresholds within the preterm and postterm gestational age ranges. This updated US-based birth weight for gestational age reference corrects for likely errors in gestational age dating and allows for the calculation of categorical and continuous measures of birth size.


Acyclovir. Pharmacology and clinical experience.
Acyclovir is a selective antiherpes agent. Its selective activity is by exploitation of differences between the herpes-specific and host cell enzymes. The major route of elimination of acyclovir is via the kidney by glomerular filtration and renal tubular secretion. In immunocompromised patients, acyclovir is effective for the prophylaxis and therapy of herpes simplex virus infections and for the treatment of herpes zoster and varicella. In nonimmunocompromised patients, acyclovir is beneficial in the therapy of herpes keratoconjunctivitis and in the therapy of genital herpes, with maximal benefit seen in those patients who have primary genital herpes. There is some benefit from acyclovir administration in patients with herpes zoster who are otherwise normal. The toxic reactions to acyclovir consist of phlebitis, local irritation, and occasionally, reversible renal dysfunction. Dose reduction is recommended in patients with impaired renal function.


Donor human milk for preterm infants: current evidence and research directions.
The Committee on Nutrition of the European Society for Pediatric Gastroenterology, Hepatology, and Nutrition aims to document the existing evidence of the benefits and common concerns deriving from the use of donor human milk (DHM) in preterm infants. The comment also outlines gaps in knowledge and gives recommendations for practice and suggestions for future research directions. Protection against necrotizing enterocolitis is the major clinical benefit deriving from the use of DHM when compared with formula. Limited data also suggest unfortified DHM to be associated with improved feeding tolerance and with reduced cardiovascular risk factors during adolescence. Presence of a human milk bank (HMB) does not decrease breast-feeding rates at discharge, but decreases the use of formula during the first weeks of life. This commentary emphasizes that fresh own mother's milk (OMM) is the first choice in preterm infant feeding and strong efforts should be made to promote lactation. When OMM is not available, DHM is the recommended alternative. When neither OMM nor DHM is available, preterm formula should be used. DHM should be provided from an established HMB, which follows specific safety guidelines. Storage and processing of human milk reduces some biological components, which may diminish its health benefits. From a nutritional point of view, DHM, like HM, does not meet the requirements of preterm infants, necessitating a specific fortification regimen to optimize growth. Future research should focus on the improvement of milk processing in HMB, particularly of heat treatment; on the optimization of HM fortification; and on further evaluation of the potential clinical benefits of processed and fortified DHM.


Fluoride exposure confers NRF2 activation in hepatocyte through both canonical and non-canonical signaling pathways.
Due to the high abundance in the Earth's crust and industrial application, fluoride is widely present in our living environment. However, excessive fluoride exposure causes toxicity in different organs. As the most important detoxification and excretion organ, liver is more easily involved in fluoride toxicity than other organs, and oxidative stress is considered as the key mechanism related with fluoride hepatotoxicity. In this study, we mainly investigated the role of nuclear factor erythroid-derived 2-like 2 (NRF2, a core transcription factor in oxidative stress) in fluoride exposure-induced hepatotoxicity as well as the related mechanism. Herein, liver cells (BNL CL.2) were treated with fluoride in different concentrations. The hepatotoxicity and NRF2 signaling pathway were analyzed respectively. Our results indicated that excessive fluoride (over 1 mM) resulted in obvious toxicity in hepatocyte and activated NRF2 and NRF2 target genes. The increased ROS generation after fluoride exposure suppressed KEAP1-induced NRF2 ubiquitylation and degradation. Meanwhile, fluoride exposure also led to blockage of autophagic flux and upregulation of p62, which contributed to activation of NRF2 via competitive binding with KEAP1. Both pharmaceutical activation and genetic activation of NRF2 accelerated fluoride exposure-induced hepatotoxicity. Thus, the upregulation of NRF2 in hepatocyte after fluoride exposure can be regarded as a cellular self-defense, and NRF2-KEAP1 system could be a novel molecular target against fluoride exposure-induced hepatotoxicity.


Evaluation of left ventricular function in coronary bypass surgery.
To evaluate non-invasive indexes measuring systolic and diastolic ventricular function. Eleven coronary artery bypass grafting (CABG) patients were investigated in order to assess the ability of preoperative ejection fraction (EF) and end diastolic pressure (EDP) to predict left ventricular function determined non-invasively at surgery. End-systolic elastance (Ees) was assessed perioperatively using transoesophageal echocardiographic area estimation and arterial pressure monitoring during preload variations (caval balloon). Diastolic function was evaluated using three different echo/Doppler indexes. EF correlated positively to Ees (r = 0.69, p = 0.03). No correlations were found between EDP and the perioperative diastolic indexes. Ees fell from pre-bypass to post-bypass (from 9.0 +/- 2.7 to 4.7 +/- 1.7 mmHg/cm2, mean +/- SD, p < 0.001), but no alterations in diastolic parameters occurred. A positive correlation was found between preoperative EF and Ees at surgery. The semi-invasive Ees detected a systolic "stunning" after cardiopulmonary bypass and is promising as a surveillance tool for left ventricular perioperative function and treatment. No correlations between preoperative EDP and non-invasive diastolic indexes were found, and assessment of perioperative diastolic function needs further refinement.


Immunohistological classification of ionocytes in the external gills of larval Japanese black salamander, Hynobius nigrescens Stejneger.
In this cytological and immunohistological study, we clarified the localization of the membrane transporters Na(+) , K(+) -ATPase (NKA), vacuolar-type H(+) -ATPase (VHA), and epithelial sodium channel (ENaC) and distinguished ionocyte subtypes in the gill of the Japanese salamander (Hynobius nigrescens). In larvae (IY stages 43-65), NKA immunoreactivity was observed on the basolateral plasma membrane in more than 60% cells and less than 20% cells in the primary filaments and secondary lamellae of the external gills, respectively. VHA immunoreactivity was observed on the apical membrane of some epithelial cells in the secondary lamellae of the external gills. High ENaCα immunoreactivity was widely observed on the apical cell membrane of a population of squamous cells, presumably pavement cells (PVCs), and mitochondria-rich cells (MRCs), in the primary filaments and secondary lamellae of the external gills. Using double immunofluorescence microscopy, epithelial cell types involved in ionic regulation were characterized and divided into three ionocyte types: NKA-, NKA- and ENaC-, and VHA-positive cells. VHA-immunoreactive cells as well as NKA-positive cells were observed during IY stages 43-65 of the salamander larvae. During late stages of metamorphosis, NKA, VHA, and ENaCα immunoreactivities in the external gills decreased and finally disappeared during the completion of metamorphosis (IY stage 68). PVCs and MRCs in the external gills are probably involved in acid-base balance regulation and osmoregulation in urodele amphibian larvae. The results are discussed in relation to the ionocytes previously reported in fish gills and the frog skin epithelium.


Phantom-derived estimation of effective dose equivalent from X rays with and without a lead apron.
Organ dose equivalents were measured in a humanoid phantom in order to estimate effective dose equivalent (H(E)) and effective dose (E) from low-energy x rays and in the presence or absence of a protective lead apron. Plane-parallel irradiation conditions were approximated using direct x-ray beams of 76 and 104 kVp and resulting dosimetry data was adjusted to model exposures conditions in fluoroscopy settings. Values of H(E) and E estimated under-shielded conditions were compared to the results of several recent studies that used combinations of measured and calculated dosimetry to model exposures to radiologists. While the estimates of H(E) and E without the lead apron were within 0.2 to 20% of expected values, estimates based on personal monitors worn at the (phantom) waist (underneath the apron) underestimated either H(E) or E while monitors placed at the neck (above the apron) significantly overestimated both quantities. Also, the experimentally determined H(E) and E were 1.4 to 3.3 times greater than might be estimated using recently reported "two-monitor" algorithms for the estimation of effective dose quantities. The results suggest that accurate estimation of either H(E) or E from personal monitors under conditions of partial body exposures remains problematic and is likely to require the use of multiple monitors.


Exploring the nature of therapeutic massage bodywork practice.
Research on therapeutic massage bodywork (TMB) continues to expand, but few studies consider how research or knowledge translation may be affected by the lack of uniformly standardized competencies for most TMB therapies, by practitioner variability from training in different forms of TMB, or from the effects of experience on practice. This study explores and describes how TMB practitioners practice, for the purpose of improving TMB training, practice, and research. PARTICIPANTS SETTING: 19 TMB practitioners trained in multiple TMB therapies, in Alberta, Canada. Qualitative descriptive sub-analysis of interviews from a comprehensive project on the training and practice of TMB, focused on the delivery of TMB therapies in practice. TWO BROAD THEMES EMERGED FROM THE DATA: (1) every treatment is individualized, and (2) each practitioner's practice of TMB therapies evolves. Individualization involves adapting treatment to the needs of the patient in the moment, based on deliberate and unconscious responses to verbal and nonverbal cues. Individualization starts with initial assessment and continues throughout the treatment encounter. Expertise is depicted as more nuanced and skilful individualization and treatment, evolved through experience, ongoing training, and spontaneous technique exploration. Practitioners consider such individualization and development of experience desirable. Furthermore, ongoing training and experience result in therapy application unique to each practitioner. Most practitioners believed they could not apply a TMB therapy without influence from other TMB therapies they had learned. There are ramifications for research design, knowledge translation, and education. Few practitioners are likely able to administer treatments in the same way, and most would not like to practice without being able to individualize treatment. TMB clinical studies need to employ research methods that accommodate the complexity of clinical practice. TMB education should facilitate the maturation of practice skills and self-reflection, including the mindful integration of multiple TMB therapies.


A metal ion-coordinated DNA probe for sensitive fluorescence detection of metallothionein via a dual nucleic acid amplification strategy.
Sensitively monitoring metallothionein (MT), a heavy metal-binding protein with substantial cysteine content, is of significance for evaluating heavy metal poisoning in both humans and animals. Based on a new metal ion-coordinated DNA probe and the heavy metal ion binding capability of MT, as well as the substantial signal enhancement of the hybridization chain reaction (HCR) and rolling circle amplification (RCA), we demonstrate a highly sensitive fluorescence MT detection assay. MT binds the metal ions in the hairpin structured, metal ion-coordinated DNA probe to switch its hairpin structure into ssDNA, which triggers subsequent RCA reactions and HCRs to open plenty of fluorescently quenched signal hairpins to exhibit drastically amplified fluorescence recovery for assaying MT down to 0.58 nM within a dynamic range of 1-320 nM. In addition, the investigation of low contents of MT in diluted human serum by such an assay has also been verified, indicating its promising application potential for diagnosing heavy metal poisoning.


Probing pH-dependent functional elements in proteins: modification of carboxylic acid pairs in Trichoderma reesei cellobiohydrolase Cel6A.
Two carboxylic acid side chains can, depending on their geometry and environment, share a proton in a hydrogen bond and form a carboxyl-carboxylate pair. In the Trichoderma reesei cellobiohydrolase Cel6A structure, five carboxyl-carboxylate pairs are observed. One of these pairs (D175-D221) is involved in catalysis, and three other pairs are found in, or close to the two surface loops covering the active site tunnel of the catalytic domain. To stabilize Cel6A at alkaline pH values, where deprotonation of the carboxylic acids leads to repulsion of their side chains, we designed two mutant enzymes. In the first mutant, one carboxyl-carboxylate pair (E107-E399) was replaced by a corresponding amide-carboxylate pair (Q107-E399), and in the second mutant, all three carboxyl-carboxylate pairs (E107-E399, D170-E184, and D366-D419) were mutated in a similar manner. The unfolding studies using both intrinsic tryptophan fluorescence and far-ultraviolet circular dichroism spectroscopy at different pH values demonstrate that the unfolding temperature (T(m)) of both mutants has changed, resulting in destabilization of the mutant enzymes at acidic pH and stabilization at alkaline pH. The effect of stabilization seems additive, as a Cel6A triple mutant is the most stable enzyme variant. This increased stability is also reflected in the 2- or 4-fold increased half-life of the two mutants at alkaline pH, while the catalytic rate on cellotetraose (at t = 0) has not changed. Increased operational stability at alkaline pH was also observed on insoluble cellulosic substrates. Local conformational changes are suggested to take place in the active site loops of Cel6A wild-type enzyme at elevated pHs (pH 7), affecting to the end-product spectrum on insoluble cellulose. The triple mutant does not show such pH-dependent behavior. Overall, our results demonstrate that carboxyl-carboxylate pair engineering is a useful tool to alter pH-dependent protein behavior.


Ocular Surface Homeostasis After Scleral Lens Usage.
The aim of this prospective study is to examine the effects of 5 hours of well-fitted, mini-scleral contact lens (mini-SL) wear on the tear film cytokine expression in healthy eyes. Twenty-three healthy participants were included in the study. One eye of each participant was selected at random, and a mini-SL measuring 16.5 mm in diameter was fitted by an experienced contact lens specialist. The contact lens remained in place for 5 hours. Precorneal tear fluid was collected using capillary tubes at three different time points: baseline before SL insertion (T0), after 5 hours of SL wear (T1), and 3 hours after SL removal (T2). The concentration of 40 inflammatory cytokines at the three different time points was determined using multiplex bead assay. Mini-scleral lens wear did not result in significant changes in the cytokine-to-protein ratio after 5 hours of wear on a healthy eye. Although a well-fitted mini-SL reduces the rate at which the precorneal tear film is refreshed, 5 hours of lens wear did not appear to significantly affect the tears cytokine-to-protein ratio, suggesting that scleral lenses have minimal impact on corneal cytokine expression.


Pharmacogenomic implications of the differential distribution of CYP2C9 metabolic phenotypes among Latin American populations.
The CYP2C9 gene encodes the major drug metabolism enzyme CYP2C9. This gene is highly polymorphic, and no-function (CYP2C9*3) plus decreased function (CYP2C9*2, *5, *8 and *11) star alleles (haplotypes) are commonly used to predict CYP2C9 metabolic phenotypes. This study explores the pharmacogenomic implications of the differential distribution of genotype-predicted CYP2C9 phenotypes across Latin American populations. Data from 1,404 individuals from the South American countries Brazil, Colombia and Peru, from Puerto Rico in the Caribbean and from persons with Mexican ancestry living in North America were analysed. The results showed that the distribution of CYP2C9 alleles and diplotypes, and diplotype-predicted CYP2C9 phenotypes vary significantly across the distinct country cohorts, as well as among self-identified White, Brown and Black Brazilians. Differences in average proportions of biogeographical ancestry across the study groups, especially Native American and African ancestry, are the likely explanation for these results. The differential distribution of genotype-predicted CYP2C9 phenotypes has potentially clinically-relevant pharmacogenomic implications, through its influence on the proportion of individuals at high risk for adverse response to medications that are CYP2C9 substrates, the proportion on individuals with CPIC therapeutic recommendations for dosing and choice of nonsteroidal antinflammatory drugs (NSAIDs) and the number of individuals that need to be genotyped in order to prevent adverse effects of NSAIDs. Collectively, these findings are likely to impact the perceived benefits, cost-effectiveness and clinical adoption of pharmacogenomic screening for drugs that are predominantly metabolized by CYP2C9.


Molecular basis of vitamin E action: tocotrienol modulates 12-lipoxygenase, a key mediator of glutamate-induced neurodegeneration.
Vitamin E is a generic term for tocopherols and tocotrienols. This work is based on our striking evidence that, in neuronal cells, nanomolar concentrations of alpha-tocotrienol, but not alpha-tocopherol, block glutamate-induced death by suppressing early activation of c-Src kinase (Sen, C. K., Khanna, S., Roy, S., and Packer, L. (2000) J. Biol. Chem. 275, 13049-13055). This study on HT4 and immature primary cortical neurons suggests a central role of 12-lipoxygenase (12-LOX) in executing glutamate-induced neurodegeneration. BL15, an inhibitor of 12-LOX, prevented glutamate-induced neurotoxicity. Moreover, neurons isolated from 12-LOX-deficient mice were observed to be resistant to glutamate-induced death. In the presence of nanomolar alpha-tocotrienol, neurons were resistant to glutamate-, homocysteine-, and l-buthionine sulfoximine-induced toxicity. Long-term time-lapse imaging studies revealed that neurons and their axo-dendritic network are fairly motile under standard culture conditions. Such motility was arrested in response to glutamate challenge. Tocotrienol-treated primary neurons maintained healthy growth and motility even in the presence of excess glutamate. The study of 12-LOX activity and metabolism revealed that this key mediator of glutamate-induced neurodegeneration is subject to control by the nutrient alpha-tocotrienol. In silico docking studies indicated that alpha-tocotrienol may hinder the access of arachidonic acid to the catalytic site of 12-LOX by binding to the opening of a solvent cavity close to the active site. These findings lend further support to alpha-tocotrienol as a potent neuroprotective form of vitamin E.


Oxprenolol in the treatment of anxiety due to environmental stress.
The author conducted two trial studies to evaluate the use of oxprenolol in the treatment of symptoms resulting from environmental stress. Results indicated that on a three-times-a-day regimen, 80 mg of oxprenolol was superior to 20 mg of oxprenolol and equally as effective as 5 mg of diazepam. The author discusses the benefit of beta-blocking drugs and points to the need for further studies.


Do seasonal patterns of rat snake (Pantherophis obsoletus) and black racer (Coluber constrictor) activity predict avian nest predation?
Avian nest success often varies seasonally and because predation is the primary cause of nest failure, seasonal variation in predator activity has been hypothesized to explain seasonal variation in nest success. Despite the fact that nest predator communities are often diverse, recent evidence from studies of snakes that are nest predators has lent some support to the link between snake activity and nest predation. However, the strength of the relationship has varied among studies. Explaining this variation is difficult, because none of these studies directly identified nest predators, the link between predator activity and nest survival was inferred. To address this knowledge gap, we examined seasonal variation in daily survival rates of 463 bird nests (of 17 bird species) and used cameras to document predator identity at 137 nests. We simultaneously quantified seasonal activity patterns of two local snake species (N = 30 individuals) using manual (2136 snake locations) and automated (89,165 movements detected) radiotelemetry. Rat snakes (Pantherophis obsoletus), the dominant snake predator at the site (~28% of observed nest predations), were most active in late May and early June, a pattern reported elsewhere for this species. When analyzing all monitored nests, we found no link between nest predation and seasonal activity of rat snakes. When analyzing only nests with known predator identities (filmed nests), however, we found that rat snakes were more likely to prey on nests during periods when they were moving the greatest distances. Similarly, analyses of all monitored nests indicated that nest survival was not linked to racer activity patterns, but racer-specific predation (N = 17 nests) of filmed nests was higher when racers were moving the greatest distances. Our results suggest that the activity of predators may be associated with higher predation rates by those predators, but that those effects can be difficult to detect when nest predator communities are diverse and predator identities are not known. Additionally, our results suggest that hand-tracking of snakes provides a reliable indicator of predator activity that may be more indicative of foraging behavior than movement frequency provided by automated telemetry systems.


Global freshwater resources: soft-path solutions for the 21st century.
Twentieth-century water policies relied on the construction of massive infrastructure in the form of dams, aqueducts, pipelines, and complex centralized treatment plants to meet human demands. These facilities brought tremendous benefits to billions of people, but they also had serious and often unanticipated social, economical, and ecological costs. Many unsolved water problems remain, and past approaches no longer seem sufficient. A transition is under way to a "soft path" that complements centralized physical infrastructure with lower cost community-scale systems, decentralized and open decision-making, water markets and equitable pricing, application of efficient technology, and environmental protection.


Detection of human rhinovirus C viral genome in blood among children with severe respiratory infections in the Philippines.
Human rhinovirus (HRV) C was recently identified as the third species of HRV using a molecular technique. Infections caused by previously identified HRVs (A and B) are thought to be limited to the respiratory tract; however, pathogenesis of HRVC is still largely unknown. A total of 816 nasopharyngeal swabs from hospitalized children with severe respiratory infections in the Philippines (May 2008-May 2009) were tested for HRV by reverse transcription polymerase chain reaction (RT-PCR), and 243 samples (29.8%) were positive for HRV. Among these patients, serum samples were also tested to determine whether specific HRV species were associated with viremia. Only 30 serum samples (12.3%) were positive for HRV. However, the HRV positive rates were different among HRV species, 3% (4/135) for HRVA, 0% (0/25) for HRVB, and 31% (26/83) for HRVC, and were the highest on 2 days after the onset of symptoms. These results suggest that HRVC may have a different pathogenicity and can more commonly cause viremia than HRVA and HRVB. Serum positive rates for HRV are affected by age, i.e., higher positive rates for those aged 1 year or more. HRVC that were detected from serum exhibited the same level of sequence diversity as those positive only for nasopharyngeal samples in phylogenetic analysis. However, all HRVA which were detected from serum were clustered in a monophyletic clade based on their 5' non-coding region (NCR) sequences, which is closely related with a certain HRVC genotype (A2) in 5'-NCR. This finding suggests that the 5'NCR region may be associated with viremia.


DNA methylation studies on imprinted loci in a male monozygotic twin pair discordant for Beckwith-Wiedemann syndrome.
Beckwith–Wiedemann syndrome (BWS) is one of the most prevalent congenital disorders predominantly caused by epigenetic alterations. Here we present an extensive case study of a monozygotic monochorionic male twin pair discordant for BWS. Our analysis allows to correlate BWS symptoms, like a protruding tongue, indented ears and transient neonatal hypoglycaemia, to an abnormal methylation at the KvDMR1. DNAs extracted from peripheral blood, skin fibroblasts, saliva and buccal swab of both twins, their sister and parents were analysed at 11 differentially methylated regions (DMRs) including all four relevant DMRs of the BWS region. The KvDMR1 was exclusively found to be hypomethylated in all cell types of the affected BWS twin, while the unaffected twin and the relatives showed normal methylation in fibroblasts, buccal swab and saliva DNA. Interestingly, the twins share a common blood-specific hypomethylation phenotype most probably caused by a feto-fetal transfusion between both twins. Because microsatellite analysis furthermore revealed a normal biparental karyotype for chromosome 11, our results point to an exclusive correlation of the observed BWS symptoms to locally restricted epimutations at the KvDMR1 of the maternal chromosome.


Switching the in vitro tRNA usage of HIV-1 by simultaneous adaptation of the PBS and PAS.
Reverse transcription of the HIV-1 RNA genome is primed by the cellular tRNA(lys3) molecule that anneals to a complementary sequence in the viral genome, the primer-binding site (PBS). Additional interactions between the tRNA primer and the viral RNA were proposed to play a role in reverse transcription. We recently identified an 8-nt element in the U5 region upstream of the PBS that is critical for initiation and processive elongation of reverse transcription. This motif was termed the primer activation signal (PAS), and is proposed to interact with the "antiPAS sequence" in the TphiC arm of tRNA(lys3). In this study, we demonstrate that the efficiency of initiation of reverse transcription can be modulated by PAS mutations that strengthen or weaken the interaction with antiPAS. These results provide further evidence for a direct base-pairing interaction between the PAS in the viral RNA and the antiPAS in the tRNA(lys3) molecule. A broad phylogenetic survey indicated that a PAS element is present in all retroviral RNA genomes, suggesting that the process of reverse transcription is regulated by a common mechanism in all retroviridae. It has proven very difficult to change the identity of the tRNA primer for HIV-1 reverse transcription by changing the PBS sequence. Using in vitro reverse transcription assays, we demonstrate that the identity of the priming tRNA species can be switched by simultaneous alteration of the PBS and PAS motifs to accommodate a new tRNA primer. These results indicate that the PAS-antiPAS interaction is important for both primer selection and efficient reverse transcription.


Factors associated with 30-day ventriculoperitoneal shunt failure in pediatric and adult patients.
OBJECTIVE The aim of this study was to provide a comprehensive benchmark of 30-day ventriculoperitoneal (VP) shunt failure rates for a single institution over a 5-year study period for both adult and pediatric patients, to compare this with the results in previously published literature, and to establish factors associated with shunt failure. METHODS A retrospective database search was undertaken to identify all VP shunt operations performed in a single, regional neurosurgical unit during a 5-year period. Data were collected regarding patient age, sex, origin of hydrocephalus, and whether the shunt was a primary or secondary shunt. Operative notes were used to ascertain the type of valve inserted, which components of the shunt were adjusted/replaced (in revision cases), level of seniority of the most senior surgeon who participated in the operation, and number of surgeons involved in the operation. Where appropriate and where available, postoperative imaging was assessed for grade of shunt placement, using a recognized grading system. Univariate and multivariate models were used to establish factors associated with early (30-day) shunt failure. RESULTS Six hundred eighty-three VP shunt operations were performed, of which 321 were pediatric and 362 were adult. The median duration of postoperative follow-up for nonfailed shunts (excluding deaths) was 1263 days (range 525-2226 days). The pediatric 30-day shunt failure rates in the authors' institution were 8.8% for primary shunts and 23.4% for revisions. In adults, the 30-day shunt failure rates are 17.7% for primary shunts and 25.6% for revisions. In pediatric procedures, the number of surgeons involved in the operating theater was significantly associated with shunt failure rate. In adults, the origin of hydrocephalus was a statistically significant variable. Primary shunts lasted longer than revision shunts, irrespective of patient age. CONCLUSIONS A benchmark of 30-day failures is presented and is consistent with current national databases and previously published data by other groups. The number of surgeons involved in shunt operations and the origin of the patient's hydrocephalus should be described in future studies and should be controlled for in any prospective work. The choice of shunt valve was not a significant predictor of shunt failure. Most previous studies on shunts have concentrated on primary shunts, but the high rate of early shunt failure in revision cases (in both adults and children) is perhaps where future research efforts should be concentrated.


Cross-linking between cytochrome c3 and flavodoxin from Desulfovibrio gigas.
Tetraheme cytochrome c3 (13 kDa) and flavodoxin (16 kDa), are small electron transfer proteins that have been used to mimic, in vitro, part of the electron-transfer chain that operates between substract electron donors and respiratory electron acceptors partners in Desulfovibrio species (Palma, N., Moura, I., LeGall, J., Van Beeumen, J., Wampler, J., Moura, J. J. G. (1994) Biochemistry 33, 6394-6407). The electron transfer between these two proteins is believed to occur through the formation of a specific complex where electrostatic interaction is the main driving force (Stewart, D., LeGall, J., Moura, I., Moura, J.J.G., Peck, H.D., Xavier, A.V., Weiner, P.K. and Wampler, J.E. (1988) Biochemistry 27, 2444-2450, Stewart, D., LeGall, J., Moura, I., Moura, J.J.G., Peck, H.D., Xavier, A.V., Weiner, P., Wampler, J. (1989) Eur. J. Biochem. 185, 695-700). In order to obtain structural information of the pre-complex, a covalent complex between the two proteins was prepared. A water-soluble carbodiimide [EDC (1-ethyl-3(3 dimethylaminopropyl) carbodiimide hydrochloride] was used for the cross linking reaction. The reaction was optimized varying a wide number of experimental parameters such as ionic strength, protein and cross linker concentration, and utilization of different cross linkers and reaction time between the crosslinker and proteins.


Population Screening for Inherited Predisposition to Breast and Ovarian Cancer.
The discovery of genes underlying inherited predisposition to breast and ovarian cancer has revolutionized the ability to identify women at high risk for these diseases before they become affected. Women who are carriers of deleterious variants in these genes can undertake surveillance and prevention measures that have been shown to reduce morbidity and mortality. However, under current strategies, the vast majority of women carriers remain undetected until they become affected. In this review, we show that universal testing, particularly of the BRCA1 and BRCA2 genes, fulfills classical disease screening criteria. This is especially true for BRCA1 and BRCA2 in Ashkenazi Jews but is translatable to all populations and may include additional genes. Utilizing genetic information for large-scale precision prevention requires a paradigmatic shift in health-care delivery. To address this need, we propose a direct-to-patient model, which is increasingly pertinent for fulfilling the promise of utilizing personal genomic information for disease prevention.


An intramolecular charge transfer and excited state intramolecular proton transfer based fluorescent probe for highly selective detection and imaging of formaldehyde in living cells.
Formaldehyde (FA), as a reactive carbonyl species, is endogenously generated in various biological processes. Abnormal levels of FA could lead to various cellular dysfunction and pathological conditions. Here, we develop a new activatable fluorescent probe for highly selective visualization of FA in living cells. Our probe (Naph-1) is designed using a naphthalene derivative as the fluorophore and hydrazone as a recognition site for FA. Naph-1 is essentially nonemissive. After reacting with FA, the amine moiety is converted into a Schiff base with electron-withdrawing ability and the fluorescence is simultaneously turned on due to synergetic intramolecular charge transfer and favoured excited state intramolecular proton transfer effects. Naph-1 exhibits a large Stokes shift upon reaction with FA. Furthermore, it possesses high selectivity and superior sensitivity toward FA with an estimated limit of detection of 0.35 μM. Moreover, Naph-1 is also successfully applied to image both endogenous and exogenous formaldehyde in living cells. These features demonstrate that Naph-1 holds great potential in the detection and imaging of formaldehyde in biological systems.


Restriction enzyme digestion of hemimethylated DNA.
Hemimethylated duplex DNA of the bacteriophage phi X 174 was synthesized using primed repair synthesis is in vitro with E. coli DNA polymerase I followed by ligation to produce the covalently closed circular duplex (RFI). Single-stranded phi X DNA was used as a template, a synthetic oligonucleotide as primer and 5-methyldeoxycytidine-5'-triphosphate (5mdCTP) was used in place of dCTP. The hemimethylated product was used as substrate for cleavage by various restriction enzymes. Out of the 17 enzymes tested, only 5 (BstN I, Taq I, Hinc II, Hinf I and Hpa I) cleaved the hemimethylated DNA. Two enzymes (Msp I and Hae III) were able to produce nicks on the unmethylated strand of the cleavage site. Msp I, which is known to cleave at CCGG when the internal cytosine residue is methylated, does not cleave when both cytosines are methylated. Another enzyme, Apy I, cleaves at the sequence CCTAGG when the internal cytosine is methylated, but is inactive on hemimethylated DNA in which both cytosines are methylated. Hemimethylated molecules should be useful for studying DNA methylation both in vivo and in vitro.


In vitro bioelectric properties of bronchial epithelium from transplanted lungs in recipients with cystic fibrosis.
BACKGROUND--Bronchial epithelial function after heart-lung transplantation (HLT) for cystic fibrosis (CF) may be affected by the original disease as well as other factors such as prolonged organ ischaemic time, the interruption of bronchial arterial and lymphatic supply, infection, rejection, and cyclosporin. In vitro measurement of the bioelectric properties of the bronchial mucosal lining may be an effective means of characterising the mucosal function of the lung allografts in response to pharmacological agents. METHODS--Bronchial mucosal tissues from explanted native lungs of CF and non-CF patients at transplantation were used to assess the possible application of a mini-Ussing chamber. With this technique, the bioelectric responses of bronchial mucosal biopsies from six patients with CF, one patient with congenital heart disease, four with primary pulmonary hypertension, and one with emphysema, all after HLT, were studied. The bioelectric and pharmacological responses of biopsies of bronchial mucosa from patients after HLT were compared with biopsies from non-CF non-HLT subjects. RESULTS--The altered bioelectric properties of CF tissues could be detected by the mini-Ussing chamber technique. The basal bioelectric values and the responses to amiloride and isoprenaline in CF patients were not different from those in non-CF patients two years after HLT. No significant difference in the basal bioelectric properties and responses to amiloride and isoprenaline was found between HLT recipients and non-CF non-HLT subjects. CONCLUSIONS--The mini-Ussing chamber is an effective means of characterising the typical CF bioelectric defect which was not found in the transplanted lungs of CF patients up to two years after HLT. Furthermore, values were unaltered in comparison with non-transplanted lungs, suggesting that bronchial epithelial function is maintained after HLT.


Septohippocampal cholinergic neurons are regulated trans-synaptically by endorphin and corticotropin neuropeptides.
The content of acetylcholine (ACh) in nerve terminals or the dorsal hippocampus was examined after intraventricular, intraseptal, or intrahippocampal administration of a variety of endorphin/corticotropin neuropeptides. beta-Lipotropin, alpha-endorphin, gamma-endorphin, alpha-melanotropin, beta-melanotropin, adrenocorticotropin-1-39 (ACTH1-39), and ACTH4-10 (1, 3, 10, or 30 micrograms each) did not affect levels of ACh in the hippocampus 30 min after injection into the lateral ventricle. beta-Endorphin, administered intraventricularly (1, 3, 10, or 30 micrograms) or intraseptally (1 microgram), increased levels of ACh, while ACTH1-24, injected similarly, decreased levels of the neurotransmitter. ACh concentrations remained unchanged after direct application of beta-endorphin or ACTH1-24 (1, 3, 10, of 30 micrograms each) into Ammon's horn. Acute unilateral transection of the fimbria/superior fornix resulted in a time-related decrease in hippocampal ACh concentrations. Levels of ACh did not change 1 hr after transection; however, concentrations of hippocampal ACh decreased significantly 1 d or 1 week after deafferentation. ACh levels in the contralateral hippocampus remained unaffected at all times tested. Fimbrial transection blocked fully both endorphin- and corticotropin-induced changes in hippocampal ACh after the neuropeptides were injected into the lateral ventricle or the septal region. Naloxone, which, after subcutaneous (1 mg/kg) or intraventricular (100 micrograms) injection alone, failed to change levels of hippocampal ACh, antagonized the effects of intraventricular or intraseptal beta-endorphin or ACTH1-24 or hippocampal ACh levels. The results suggest a site of endorphin/corticotropin receptor interaction at the level of cholinergic cell bodies in the septal region for regulating the activity of septohippocampal cholinergic neurons.


Homology modeling and antagonist binding site study of the human histamine H2 receptor.
On the basis of the high resolution crystal structures of bovine rhodopsin, human beta2-adrenergic receptor and human A(2a) adenosine receptor, three-dimensional structure of the human histamine H2 receptor (HHR2) was developed by homology modeling. Results of the evaluations suggest that a high quality homology model for HHR2 has been obtained. Three antagonists, cimetidine, ranitidine and nizatidine, were applied to binding site study with this model through molecular docking, molecular dynamics simulations and the molecular mechanics-Poisson-Boltzmann surface area calculations. One aspartic acid, Asp98 in transmembrane domain 7 (TM3), has been identified as major contributors to ligand binding by H-bond interactions. Asn159 in TM4 and Asp186 in TM5 are of great importance in stabilizing HHR2- antagonist complexes. Two hydrophobic sites especially two residues, Val99 in TM3 and Phe254 in TM6, were identified to be essential for their strong hydrophobic interactions with antagonists. The findings reported here are in agreement with available experimental mutagenesis data. This study should be very helpful for a better understanding of the action mode of the antagonist and for guiding further drug design and mutagenesis studies.


[A Turkish man with Behçet disease and recurrent acute abdomen].
Following a total hip prosthesis, a 52-year-old Turkish man with Behçet's disease developed persistent ileus due to intestinal perforations. Resection of the intestine was followed by new perforations, resulting in resection of another portion of the intestine and the start of immunosuppressive medication. Following the 5th resection for a perforated ulcer in the space of two weeks, an ileostomy was performed. One year later the stoma could be eliminated and the patient remained symptom-free. Behçet's disease is relatively rare in the Netherlands. However, the gastrointestinal complications may be life-threatening. Surgical intervention is then often necessary. Complications and recurrences are frequent.


A cautionary note on data sources for evidence-based clinical decisions: warfarin and stroke prevention.
Stroke risk in nonvalvular atrial fibrillation can be reduced by warfarin or aspirin; the choice of therapy requires the assessment of risks and benefits. The authors compared methods of risk assessment and their implications for risk communication and treatment. Stroke risk was compared in 193 patients with atrial fibrillation using the Framingham equation; an atrial fibrillation-specific Framingham equation; the Congestive heart failure, Hypertension, Age, Diabetes and Stroke (CHADS(2)) score; the Stroke Prevention and Atrial Fibrillation (SPAF) scheme; and the Scottish Intercollegiate Guidelines Network (SIGN) guidelines. Treatment guidance from SIGN, a simple prediction rule, and a decision analytical approach was compared. In the latter, patients were classified as risk too low to benefit from warfarin if the risk of cerebral bleeding on warfarin approximated to, or exceeded, thromboembolic stroke risk reduction. Framingham equations gave lower stroke risks overall than SIGN or SPAF. CHADS(2) was intermediate. Using SIGN, warfarin would be given to all 103 patients without a history of stroke/transient ischemic attack and for whom warfarin was not contraindicated but only to 73 patients using the simple prediction rule and 48 patients using the decision analysis. Community-based cohorts give lower stroke risk estimates than CHADS(2); both give lower estimates than schemes from control groups from randomized controlled trials. Using community-derived risks would lead to fewer patients being treated with warfarin than guidance derived from randomized controlled trial controls, which may lead to many low-risk patients being treated with high-risk therapy. This raises the debate about appropriate sources of data for risk assessment to support risk communication and effective clinical decisions.


Proximal disease extension and related predicting factors in ulcerative proctitis.
Ulcerative colitis usually involves the rectum, may extend in a proximal and continuous fashion to involve varying portions of the bowel. However, the risk factors predictive of proximal extension have yet to be determined. The aim of this study was to evaluate both the natural course of disease and the risk factors influencing the proximal disease extension in ulcerative proctitis. We retrospectively analyzed 98 patients with ulcerative proctitis at the time of diagnosis who were regularly followed and underwent sigmoidoscopy or colonoscopy between January 2000 and December 2007. The mean duration of follow-up was 109.2 ± 49.5 months. A total of 27 (27.6%) patients experienced proximal disease extension. Mayo scores were significantly higher in the extension group compared with patients whose ulcerative proctitis did not extend proximally (p < 0.001). Corticosteroid use at initial diagnosis was also more frequent in the extension group (p = 0.026). In addition, chronic, continuous disease activation within 6 months of the initial diagnosis was significantly higher in the extension group (p < 0.001), as was disease relapse and the number of hospitalizations over the entire follow-up period (p < 0.001 and p = 0.002). According to multivariate analysis, disease extension after the initial diagnosis was associated with chronic disease activation, disease relapse and hospitalization (p = 0.030, p = 0.042 and p = 0.044, respectively). Increased severity of disease upon diagnosis of ulcerative proctitis was associated with a higher probability of proximal disease extension during the follow-up period. Moreover, those with disease extension were more likely to experience relapse and to be hospitalized, indicating poor prognosis.


CC chemokine receptor 5 and CXC chemokine receptor 6 expression by lung CD8+ cells correlates with chronic obstructive pulmonary disease severity.
Chronic obstructive pulmonary disease (COPD) is a progressive disease associated with a cellular inflammatory response. CD8(+) T cells are implicated in COPD pathogenesis, and their numbers significantly correlate with the degree of airflow limitation. Dendritic cells (DCs) are important sentinel immune cells, but little is known about their role in initiating and maintaining the CD8 T-cell response in COPD. To investigate the mechanisms for CD8(+) T-cell recruitment to the lung, we used resected human lung tissue to analyze chemokine receptor expression by CD8(+) T cells and chemokine production by CD1a(+) DCs. Among 11 surveyed chemokine receptors, only CC chemokine receptor (CCR5), CXC chemokine receptor (CXCR) 3, and CXCR6 correlated with COPD severity as defined by criteria from the Global Initiative for Chronic Obstructive Lung Disease. The CD8(+) T cells displayed a Tc1, CD45RA(+) effector memory phenotype. CD1a(+) DCs produced the respective ligands for CCR5 and CXCR3, CCL3 and CXCL9, and levels correlated with disease severity. CD1a(+) DCs also constitutively expressed the CXCR6 ligand, CXCL16. In conclusion, we have identified major chemokine elements that potentially mediate CD8(+) T-cell infiltration during COPD progression and demonstrated that CD1a(+) mucosal-associated DCs may sustain CD8(+) T-cell recruitment/retention. Chemokine targeting may prove to be a viable treatment approach.


Regional variation in contribution of myenteric and intramuscular interstitial cells of Cajal to generation of slow waves in mouse gastric antrum.
When intracellular recordings were made from the antral region of murine stomach, cells with three different patterns of electrical activity were detected. One group of cells generated follower potentials, the second group generated pacemaker potentials and the third group generated slow waves that consisted of primary and secondary components. Slow waves recorded in different regions of the gastric antrum had similar amplitudes but different characteristic shapes. At the greater curvature, slow waves had large initial components. Midway between the greater and lesser curvature, the amplitude of the initial component was reduced and at the lesser curvature an initial component was difficult to detect. When the distributions of myenteric (ICC-MY) and intramuscular interstitial cells of Cajal (ICC-IM) were determined, using an antibody to Kit, ICC-MY were found to be present at the greater curvature but were greatly reduced in density at the lesser curvature. In contrast, ICC-IM were found in the circular layer of each region. When recordings were made from the antrum of W/W(V) mice, which lack ICC-IM, incomplete slow waves were detected and their amplitudes fell from the greater to the lesser curvature. Again, a corresponding fall in the density of ICC-MY was detected. The observations indicate that the contribution of ICC-MY and ICC-IM to the generation of slow waves varies in different regions of the mouse gastric antrum.


Regulatory role of prostaglandin E2 in induction of cyclo-oxygenase-2 by a thromboxane A2 analogue (U46619) and basic fibroblast growth factor in porcine aortic smooth-muscle cells.
U46619, a thromboxane A2 analogue, and basic fibroblast growth factor (FGF-2) both induced the expression of the inducible cyclo-oxygenase (Cox)-2 in porcine aortic smooth-muscle cells. This induction was dose-dependent (submaximal at 300 nM for U46619 and 1 ng/ml for FGF-2) and time-dependent, with similar intensity and maximal expression at 2 h. Under these conditions, both inducers stimulated rapid activation of extracellular signal-regulated kinase (ERK2) at 5-10 min, a transient and lower intensity being induced by U46619 whereas that induced by FGF-2 was sustained (>1 h). PD98059, an inhibitor of the ERK pathway, inhibited the expression of Cox-2. In contrast, activation of Jun-N-terminal kinase (JNK1) was sustained with U46619 but poorly induced by FGF-2. Cox-2 expression induced by U46619 or FGF-2 was similarly reduced by prostaglandin (PGE2), forskolin or dibutyryl-cAMP, suggesting a regulatory effect of adenylate cyclase on Cox-2 expression. However, activation of ERK2 by FGF-2 was not affected by PGE2 whereas that of JNK1 by U46619 was inhibited, suggesting that inhibition of COX-2 expression by cAMP may be downstream of ERK2. The effects of PGE2 and forskolin on Cox-2 and phosphorylation of JNK1 were reversed with the protein kinase A inhibitor H89. In addition, endogenous PGE2 down-regulated the expression of Cox-2 by the two inducers, as stimulation of the cells in the presence of different Cox inhibitors increased the expression of the protein. Overall, these results suggest that exogenous and endogenous PGE2 exert negative inhibitory effects on Cox-2 expression mediated by stimulation of protein kinase A.


Preclinical pharmacology of perampanel, a selective non-competitive AMPA receptor antagonist.
Perampanel [2-(2-oxo-1-phenyl-5-pyridin-2-yl-1,2-dihydropyridin-3-yl)benzonitrile; E2007] is a potent, selective, orally active non-competitive AMPA receptor antagonist developed for the treatment of epilepsy. Perampanel has a 2,3'-bipyridin-6'-one core structure, distinguishing it chemically from other AMPA receptor antagonist classes. Studies in various physiological systems indicate that perampanel selectively inhibits AMPA receptor-mediated synaptic excitation without affecting NMDA receptor responses. Blocking of AMPA receptors occurs at an allosteric site that is distinct from the glutamate recognition site. Radioligand-binding studies suggest that the blocking site coincides with that of the non-competitive antagonist GYKI 52466, believed to be on linker peptide segments of AMPA receptor subunits that transduce agonist binding into channel opening. As is typical for AMPA receptor antagonists, perampanel exhibits broad-spectrum antiseizure activity in diverse animal seizure models. Perampanel has high oral bioavailability, dose-proportional kinetics, and undergoes oxidative metabolism, primarily via CYP3A4, followed by glucuronidation. The terminal half-life (t½ ) in humans is 105 h; however, in the presence of a strong CYP3A4 inducer (such as carbamazepine), the t½ can be reduced. In sum, perampanel is a selective, centrally acting, negative allosteric modulator of AMPA receptors with good oral bioavailability and favorable pharmacokinetic properties.


Nonlinear image recovery with half-quadratic regularization.
One popular method for the recovery of an ideal intensity image from corrupted or indirect measurements is regularization: minimize an objective function that enforces a roughness penalty in addition to coherence with the data. Linear estimates are relatively easy to compute but generally introduce systematic errors; for example, they are incapable of recovering discontinuities and other important image attributes. In contrast, nonlinear estimates are more accurate but are often far less accessible. This is particularly true when the objective function is nonconvex, and the distribution of each data component depends on many image components through a linear operator with broad support. Our approach is based on an auxiliary array and an extended objective function in which the original variables appear quadratically and the auxiliary variables are decoupled. Minimizing over the auxiliary array alone yields the original function so that the original image estimate can be obtained by joint minimization. This can be done efficiently by Monte Carlo methods, for example by FFT-based annealing using a Markov chain that alternates between (global) transitions from one array to the other. Experiments are reported in optical astronomy, with space telescope data, and computed tomography.


A Quantitative Framework to Study Potential Benefits and Harms of Multi-Cancer Early Detection Testing.
Multi-cancer tests offer screening for multiple cancers with one blood draw, but the potential population impact is poorly understood. We formulate mathematical expressions for expected numbers of: (i) individuals exposed to unnecessary confirmation tests (EUC), (ii) cancers detected (CD), and (iii) lives saved (LS) given test performance, disease incidence and mortality, and mortality reduction. We add colorectal, liver, lung, ovary, and pancreatic cancer to a test for breast cancer, approximating prevalence at ages 50, 60, or 70 using incidence over the next 5 years and mortality using corresponding probabilities of cancer death over 15 years in the Surveillance, Epidemiology, and End Results registry. EUC is overwhelmingly determined by specificity. For a given specificity, EUC/CD is most favorable for higher prevalence cancers. Under 99% specificity and sensitivities as published for a 50-cancer test, EUC/CD is 1.1 for breast + lung versus 1.3 for breast + liver at age 50. Under a common mortality reduction associated with screening, EUC/LS> is most favorable when the test includes higher mortality cancers (e.g., 19.9 for breast + lung vs. 30.4 for breast + liver at age 50 assuming a common 10% mortality reduction). Published multi-cancer test performance suggests a favorable tradeoff of EUC to CD, yet the full burden of unnecessary confirmations will depend on the posttest work-up protocol. Harm-benefit tradeoffs will be improved if tests prioritize more prevalent and/or lethal cancers for which curative treatments exist. The population impact of multi-cancer testing will depend not only on test performance but also on disease characteristics and efficacy of early treatment.See related commentary by Duffy and Sasieni, p. 3.


Optimization of geometrical calibration in pinhole SPECT.
Previously, we developed a method to determine the acquisition geometry of a pinhole camera. This information is needed for the correct reconstruction of pinhole single photon emission computed tomography images. The method uses a calibration phantom consisting of three point sources and their positions in the field of view (FOV) influence the accuracy of the geometry estimate. This paper proposes two particular configurations of point sources with specific positions and orientations in the FOV for optimal image reconstruction accuracy. For the proposed calibration setups, inaccuracies of the geometry estimate due to noise in the calibration data, only cause subresolution inaccuracies in reconstructed images. The calibration method also uses a model of the point source configuration, which is only known with limited accuracy. The study demonstrates, however, that, with the proposed calibration setups, the error in reconstructed images is comparable to the error in the phantom model.


Online Studies on Variation in Orthopedic Surgery: Computed Tomography in MPEG4 Versus DICOM Format.
The purpose of this study was to compare the observer participation and satisfaction as well as interobserver reliability between two online platforms, Science of Variation Group (SOVG) and Traumaplatform Study Collaborative, for the evaluation of complex tibial plateau fractures using computed tomography in MPEG4 and DICOM format. A total of 143 observers started with the online evaluation of 15 complex tibial plateau fractures via either the SOVG or Traumaplatform Study Collaborative websites using MPEG4 videos or a DICOM viewer, respectively. Observers were asked to indicate the absence or presence of four tibial plateau fracture characteristics and to rate their satisfaction with the evaluation as provided by the respective online platforms. The observer participation rate was significantly higher in the SOVG (MPEG4 video) group compared to that in the Traumaplatform Study Collaborative (DICOM viewer) group (75 and 43%, respectively; P < 0.001). The median observer satisfaction with the online evaluation was seven (range, 0-10) using MPEG4 video compared to six (range, 1-9) using DICOM viewer (P = 0.11). The interobserver reliability for recognition of fracture characteristics in complex tibial plateau fractures was higher for the evaluation using MPEG4 video. In conclusion, observer participation and interobserver reliability for the characterization of tibial plateau fractures was greater with MPEG4 videos than with a standard DICOM viewer, while there was no difference in observer satisfaction. Future reliability studies should account for the method of delivering images.


Spatial coherence resonance on diffusive and small-world networks of Hodgkin-Huxley neurons.
Spatial coherence resonance in a spatially extended system that is locally modeled by Hodgkin-Huxley (HH) neurons is studied in this paper. We focus on the ability of additive temporally and spatially uncorrelated Gaussian noise to extract a particular spatial frequency of excitatory waves in the medium, whereby examining the impact of diffusive and small-world network topology that determines the interactions amongst coupled HH neurons. We show that there exists an intermediate noise intensity that is able to extract a characteristic spatial frequency of the system in a resonant manner provided the latter is diffusively coupled, thus indicating the existence of spatial coherence resonance. However, as the diffusive topology of the medium is relaxed via the introduction of shortcut links introducing small-world properties amongst coupled HH neurons, the ability of additive Gaussian noise to evoke ordered excitatory waves deteriorates rather spectacularly, leading to the decoherence of the spatial dynamics and with it related absence of spatial coherence resonance. In particular, already a minute fraction of shortcut links suffices to substantially disrupt coherent pattern formation in the examined system.


Myo-Inositol Limits Kainic Acid-Induced Epileptogenesis in Rats.
Epilepsy is a severe neurological disease characterized by spontaneous recurrent seizures (SRS). A complex pathophysiological process referred to as epileptogenesis transforms a normal brain into an epileptic one. Prevention of epileptogenesis is a subject of intensive research. Currently, there are no clinically approved drugs that can act as preventive medication. Our previous studies have revealed highly promising antiepileptogenic properties of a compound-myo-inositol (MI) and the present research broadens previous results and demonstrates the long-term disease-modifying effect of this drug, as well as the amelioration of cognitive comorbidities. For the first time, we show that long-term treatment with MI: (i) decreases the frequency and duration of electrographic SRS in the hippocampus; (ii) has an ameliorating effect on spatial learning and memory deficit associated with epileptogenesis, and (iii) attenuates cell loss in the hippocampus. MI treatment also alters the expression of the glial fibrillary acidic protein, LRRC8A subunit of volume-regulated anion channels, and protein tyrosine phosphatase receptor type R, all expected to counteract the epileptogenesis. All these effects are still present even 4 weeks after MI treatment ceased. This suggests that MI may exert multiple actions on various epileptogenesis-associated changes in the brain and, therefore, could be considered as a candidate target for prevention of epileptogenesis.


Inhibitory Effect of Naphthoquinone-Tryptophan Hybrid towards Aggregation of PAP f39 Semen Amyloid.
PAP248⁻286, a 39 amino acid peptide fragment, derived from the prostatic acid phosphatase secreted in human semen, forms amyloid fibrils and facilitates the attachment of retroviruses to host cells that results in the enhancement of viral infection. Therefore, the inhibition of amyloid formation by PAP248⁻286 (termed PAP f39) may likely reduce HIV transmission in AIDS. In this study, we show that the naphthoquinone tryptophan (NQTrp) hybrid molecule significantly inhibited PAP f39 aggregation in vitro in a dose-dependent manner as observed from the ThT assay, ANS assay, and transmission electron microscopy imaging. We found that even at a sub-molar concentration of 20:1 [PAP f39:NQTrp], NQTrp could reduce >50% amyloid formation. NQTrp inhibition of PAP f39 aggregation resulted in non-toxic intermediate species as determined by the vesicle leakage assay. Isothermal titration calorimetry and molecular docking revealed that the binding of NQTrp and PAP f39 is spontaneous, and NQTrp predominantly interacts with the polar and charged residues of the peptide by forming hydrogen bonds and hydrophobic contacts with a strong binding energy. Collectively, these findings indicate that NQTrp holds significant potential as a small molecule inhibitor of semen amyloids.


Upregulation of miRNA-4776 in Influenza Virus Infected Bronchial Epithelial Cells Is Associated with Downregulation of NFKBIB and Increased Viral Survival.
Influenza A virus (IAV) infection remains a significant cause of morbidity and mortality worldwide. One key transcription factor that is activated upon IAV infection is nuclear factor Kappa B (NF-κB). NF-κB regulation involves the inhibitor proteins NF-κB inhibitor beta (NFKBIB), (also known as IκB β), which form complexes with NF-κB to sequester it in the cytoplasm. In this study, microarray data showed differential expression of several microRNAs (miRNAs) on exposure to IAV. Target scan analysis revealed that miR-4776, miR-4514 and miR-4742 potentially target NFKBIB messenger RNA (mRNA). Time-course analysis of primary bronchial epithelial cells (HBEpCs) showed that miR-4776 expression is increased within 1 h of infection, followed by its downregulation 4 h post-exposure to IAV. NFKBIB upregulation of miR-4776 correlated with a decrease in NFKBIB expression within 1 h of infection and a subsequent increase in NFKBIB expression 4 h post-infection. In addition, miRNA ago-immunoprecipitation studies and the three prime untranslated region (3' UTR) luciferase assay confirmed that miR-4776 targets NFKBIB mRNA. Furthermore, uninfected HBEpCs transfected with miR-4776 mimic showed decreased expression of NFKBIB mRNA. Overexpression of NFKBIB protein in IAV infected cells led to lower levels of IAV. Taken together, our data suggest that miRNA-4776 modulates IAV production in infected cells through NFKBIB expression, possibly through the modulation of NF-κB.