Long-term stability and performance characteristics of crystal quartz gauge at high pressures and temperatures.
Good long-term stability of high precision quartz pressure sensors is necessary for various applications ranging from pressure transient analysis to permanent monitoring systems for optimal reservoir management in the petroleum industry. A crystal quartz gauge (CQG(1 )) is a dual-mode, thickness-shear, quartz pressure sensor that has been used in oil field services for the past 8 years. High accuracy, resolution, and fast response time of this sensor enable a reliable estimate of formation permeability and oil/water interfaces in reservoirs that help reduce the overall cost of oil and gas production. The sensing resonator characteristics can be described in terms of equivalent circuit parameters (motional resistance and capacitance), resonator-Q and the short-term frequency stability of both the B- and C-modes of vibrations at various temperatures. The pressure reading errors of manufactured gauges are less than 8.89 kPa (1 psi) (plus 0.01% of the reading because of the uncertainty of the dead-weight tester). The pressure resolution is better than 20.7 Pa (0.003 psi) over a 1-s gate time. An extremely effective dynamic compensation algorithm yields corrected pressure readings with a very fast response time as short as a strain-gauge-based pressure transducer while retaining the high performance of a quartz gauge. Recent long-term stability tests of CBGs show a negligibly small drift of the order of a few tenths of 1 psi (0.1 psi=689 Pa) at 103 MPa (15 kpsi) and 175 degrees C for a period of more than 1 yr. These results confirm that the CQG characteristics exceed the demanding specifications for both the well tests and permanent monitoring systems.


A case series of fetal lymphatic malformations and a review of the literature.
Lymphatic malformations are rare benign developmental anomalies of the lymphatic system that can be diagnosed by prenatal ultrasound. Depending on their anatomical site and size, the lesions can cause a variety of aesthetic and functional deficits. Several treatment options are available, the most suitable is still under debate. The experience gained at our Centre and the review of the literature can be useful to improve prenatal counseling, that is challenging due to the heterogeneity of clinical presentation and treatment.


HIV testing and treatment with correctional populations: people, not prisoners.
Institutional policies, practices, and norms can impede the delivery of ethical standard-of-care treatment for people with HIV in correctional settings. In this commentary, we focus on the fundamental issues that must be addressed to create an ethical environment in which best medical practices can be implemented when working with correctional populations. Thus, we consider ethical issues related to access to services, patient privacy, confidentiality, informed consent for testing and treatment, and issues related to the provision of services in an institutional setting in which maintenance of security is the primary mission. Medical providers must understand and navigate the dehumanization inherent in most correctional settings, competing life demands for incarcerated individuals, power dynamics within the correctional system, and the needs of family and significant others who remain in the community.


Carcinoma of the gallbladder: a diagnostic challenge.
Since the introduction and availability of abdominal ultrasound and computed tomography, 38 patients have been admitted to the Department of Surgery, Queen Mary Hospital, University of Hong Kong with a subsequently confirmed histological diagnosis of carcinoma of the gallbladder. The majority of patients presented with abdominal pain, anorexia or weight loss, jaundice, and anaemia or an abdominal swelling. Abnormal liver function tests were found in 70-90%. Cholangiography, ultrasound and particularly computed tomography helped to confirm clinical suspicion when performed but 23 patients (61%) had an inaccurate diagnosis made before surgery. Operative intervention was only of benefit when the carcinoma was confined to the gallbladder (median survival after cholecystectomy alone 13 months; other procedures 2 months; P less than 0.01). Carcinoma of the gallbladder carries a poor prognosis but ultrasound and computed tomography can help select patients for appropriate treatment (surgery, endoscopic stenting, radiotherapy, chemotherapy) if clinical suspicion is raised.


Excretion of endogenous boldione in human urine: influence of phytosterol consumption.
Boldenone (17-hydroxy-androsta-1,4-diene-3-one, Bol) and boldione (androst-1,4-diene-3,17-dione, ADD), are currently listed as exogenous anabolic steroids by the World Anti-Doping Agency. However, it has been reported that these analytes can be produced endogenously. Interestingly, only for Bol a comment is included in the list on its potential endogenous origin. In this study, the endogenous origin of ADD in human urine was investigated, and the potential influence of phytosterol consumption was evaluated. We carried out a 5-week in vivo trial with both men (n=6) and women (n=6) and measured alpha-boldenone, beta-boldenone, boldione, androstenedione, beta-testosterone and alpha-testosterone in their urine using gas chromatography coupled to multiple mass spectrometry (GC-MS-MS). The results demonstrate that endogenous ADD is sporadically produced at concentrations ranging from 0.751 ng mL(-1) to 1.73 ng mL(-1), whereas endogenous Bol could not be proven. We also tested the effect of the daily consumption of a commercially available phytosterol-enriched yogurt drink on the presence of these analytes in human urine. Results from this study could not indicate a relation of ADD-excretion with the consumption of phytosterols at the recommended dose. The correlations between ADD and other steroids were consistently stronger for volunteers consuming phytosterols (test) than for those refraining from phytosterol consumption (control). Excretion of AED, bT and aT did not appear to be dependent on the consumption of phytosterols. This preliminary in vivo trial indicates the endogenous origin of boldione or ADD in human urine, independent on the presence of any structural related analytes such as phytosterols.


The synergic effects of quinolones and oral cephem antibiotics on Serratia marcescens.
The in-vitro effects of new quinolones and oral cephem antibiotics in various combinations on 26 strains of clinically isolated Serratia marcescens were assessed. The in-vitro activities of ofloxacin and ciprofloxacin in combination with either cefpodoxime or cefcamate were evaluated by the chequerboard titration method. These combinations showed synergistic and additive effects against most of the S. marcescens tested but no antagonism was observed. Time-kill experiments performed with two representative isolates of S. marcescens showed synergic effects with respect to the kill rates, when concentrations of quinolones and beta-lactam antibiotics, exhibiting bacteriostatic effects were combined. Exposure of S. marcescens to ciprofloxacin before the addition of cefpodoxime had no influence on the combination effect. When cefpodoxime was added first, however, the bactericidal effect was reduced. The induction of beta-lactamase by cefmetazole was enhanced by the combination of ciprofloxacin and cefmetazole. These results suggest that the synergic effects observed between new quinolones and cephem antibiotics are due to the stimulation of incorporation of cephems by combination with quinolones.


Multiple unrelated translocations in a metastatic epimyoepithelial carcinoma of the parotid gland.
We report the cytogenetic, fluorescence in-situ hybridization (FISH) and DNA flow cytometric analysis of a rare metastatic epimyoepithelial carcinoma of the parotid gland to the lung with a clinical course of 29 years. DNA content and FISH analyses of tumor and short-term culture cells showed diploid DNA content and lack of numerical chromosomal abnormalities. Immunohistochemical analysis of the short-term culture cells showed predominantly keratin positive and sparse desmin staining supporting an epithelial rather than myoepithelial origin. Cytogenetic analysis showed 46,XY karyotype with clonal translocations of t(3;22)(q13.2;q13.1), t(1;7)(q21;q22), t(8;9)(p10;p10), and t(5;6) (q35;q21). Our findings indicate that these alterations developed in a diploid stemline during tumor progression and in the epithelial component of this tumor.


Weightless experiments to probe universality of fluid critical behavior.
Near the critical point of fluids, critical opalescence results in light attenuation, or turbidity increase, that can be used to probe the universality of critical behavior. Turbidity measurements in SF6 under weightlessness conditions on board the International Space Station are performed to appraise such behavior in terms of both temperature and density distances from the critical point. Data are obtained in a temperature range, far (1 K) from and extremely close (a few μK) to the phase transition, unattainable from previous experiments on Earth. Data are analyzed with renormalization-group matching classical-to-critical crossover models of the universal equation of state. It results that the data in the unexplored region, which is a minute deviant from the critical density value, still show adverse effects for testing the true asymptotic nature of the critical point phenomena.


Cellular Protein WDR11 Interacts with Specific Herpes Simplex Virus Proteins at the trans-Golgi Network To Promote Virus Replication.
It has recently been proposed that the herpes simplex virus (HSV) protein ICP0 has cytoplasmic roles in blocking antiviral signaling and in promoting viral replication in addition to its well-known proteasome-dependent functions in the nucleus. However, the mechanisms through which it produces these effects remain unclear. While investigating this further, we identified a novel cytoplasmic interaction between ICP0 and the poorly characterized cellular protein WDR11. During an HSV infection, WDR11 undergoes a dramatic change in localization at late times in the viral replication cycle, moving from defined perinuclear structures to a dispersed cytoplasmic distribution. While this relocation was not observed during infection with viruses other than HSV-1 and correlated with efficient HSV-1 replication, the redistribution was found to occur independently of ICP0 expression, instead requiring viral late gene expression. We demonstrate for the first time that WDR11 is localized to the trans-Golgi network (TGN), where it interacts specifically with some, but not all, HSV virion components, in addition to ICP0. Knockdown of WDR11 in cultured human cells resulted in a modest but consistent decrease in yields of both wild-type and ICP0-null viruses, in the supernatant and cell-associated fractions, without affecting viral gene expression. Although further study is required, we propose that WDR11 participates in viral assembly and/or secondary envelopment. While the TGN has been proposed to be the major site of HSV-1 secondary envelopment, this process is incompletely understood, and in particular, the role of cellular TGN components in this pathway is unknown. Additionally, little is known about the cellular functions of WDR11, although the disruption of this protein has been implicated in multiple human diseases. Therefore, our finding that WDR11 is a TGN-resident protein that interacts with specific viral proteins to enhance viral yields improves both our understanding of basic cellular biology as well as how this protein is co-opted by HSV.


Ruthenium-catalyzed benzylic substitution of benzyl esters with stabilized carbon nucleophiles.
We have accomplished the ruthenium-catalyzed benzylic substitution of benzyl esters with a stabilized carbon nucleophile. A [Cp*RuCl2]2/picolinic acid catalyst system promoted the reaction of 2-naphthylmethyl-2,3,4,5,6-pentafluorobenzoates with a series of stabilized carbon nucleophiles such as malonates, β-ketoesters, and diketones to give the corresponding benzylic alkylation products in moderate to high yields. We proposed a plausible reaction mechanism that could involve a (π-benzyl)ruthenium intermediate.


Threshold received sound pressure levels of single 1-2 kHz and 6-7 kHz up-sweeps and down-sweeps causing startle responses in a harbor porpoise (Phocoena phocoena).
Mid-frequency and low-frequency sonar systems produce frequency-modulated sweeps which may affect harbor porpoises. To study the effect of sweeps on behavioral responses (specifically "startle" responses, which we define as sudden changes in swimming speed and/or direction), a harbor porpoise in a large pool was exposed to three pairs of sweeps: a 1-2 kHz up-sweep was compared with a 2-1 kHz down-sweep, both with and without harmonics, and a 6-7 kHz up-sweep was compared with a 7-6 kHz down-sweep without harmonics. Sweeps were presented at five spatially averaged received levels (mRLs; 6 dB steps; identical for the up-sweep and down-sweep of each pair). During sweep presentation, startle responses were recorded. There was no difference in the mRLs causing startle responses for up-sweeps and down-sweeps within frequency pairs. For 1-2 kHz sweeps without harmonics, a 50% startle response rate occurred at mRLs of 133 dB re 1 μPa; for 1-2 kHz sweeps with strong harmonics at 99 dB re 1 μPa; for 6-7 kHz sweeps without harmonics at 101 dB re 1 μPa. Low-frequency (1-2 kHz) active naval sonar systems without harmonics can therefore operate at higher source levels than mid-frequency (6-7 kHz) active sonar systems without harmonics, with similar startle effects on porpoises.


The promise of comparative genomics in mammals.
Dense genetic maps of human, mouse, and rat genomes that are based on coding genes and on microsatellite and single-nucleotide polymorphism markers have been complemented by precise gene homolog alignment with moderate-resolution maps of livestock, companion animals, and additional mammal species. Comparative genetic assessment expands the utility of these maps in gene discovery, in functional genomics, and in tracking the evolutionary forces that sculpted the genome organization of modern mammalian species.


Whole Genome Sequencing for the Retrospective Investigation of an Outbreak of Salmonella Typhimurium DT 8.
Salmonella enterica serovar Typhimurium DT8 is uncommon within the European Union. An increase in this phage type was reported in the summer of 2013 in the States of Jersey. A total of 21 human cases with this phage type were microbiologically confirmed. Salmonella isolates from mayonnaise made using raw eggs were also confirmed as being Salmonella Typhimurium DT8. The epidemiological investigations strongly supported a link between mayonnaise consumption and illness. Whole genome sequencing (WGS) was used to retrospectively investigate this outbreak with a view to assess the similarity between the suspect food and the human isolates and to characterise a known point source outbreak to assist in development of algorithms for outbreak detection. Sequence data showed that the outbreak associated isolates, including the food isolates, formed a tightly clustered monophyletic group, with a maximum pairwise distance of 3 single nucleotide polymorphisms. WGS data is useful in confirming the causative agent of outbreaks where food and clinical isolates are available. This dataset, comprising a known outbreak, will be useful in the development of automatic algorithms for outbreak detection.


Internal filtration, filtration fraction, and blood flow resistance in high- and low-flux dialyzers.
It was the aim to examine the fluid flow in blood and dialysate compartments of highly permeable hollow fiber dialyzers where internal filtration contributes to solute removal but where excessive filtration bears a risk of cell activation and damage. Flow characteristics of high- (HF) and low-flux (LF) dialyzers were studied in lab-bench experiments using whole bovine blood. Measurements obtained under different operating conditions and under zero net ultrafiltration were compared to theoretical calculations obtained from a mathematical model. Experimental resistances in the blood compartment were within ±2% of those calculated from the model when dialysate was used as a test fluid. With whole blood, the experimental resistances in the blood compartment were only 81.8 ± 2.8% and 83.7 ± 4.3% of those calculated for the LF and HF dialyzer, respectively. Surprisingly, measured blood flow resistance slightly but significantly decreased with increasing flow rate (p < 0.001). Mathematical modeling confirmed this decrease both in LF and HF dialyzers which was accompanied by a concomitant decrease in internal filtration fraction, while overall internal filtration increased. The increase in internal filtration when increasing blood flow is associated with a beneficial reduction in internal filtration fraction. Concerns of increased hemoconcentration when increasing blood flow therefore appear to be unwarranted.


Are Lockdowns Effective in Managing Pandemics?
The present coronavirus crisis caused a major worldwide disruption which has not been experienced for decades. The lockdown-based crisis management was implemented by nearly all the countries, and studies confirming lockdown effectiveness can be found alongside the studies questioning it. In this work, we performed a narrative review of the works studying the above effectiveness, as well as the historic experience of previous pandemics and risk-benefit analysis based on the connection of health and wealth. Our aim was to learn lessons and analyze ways to improve the management of similar events in the future. The comparative analysis of different countries showed that the assumption of lockdowns' effectiveness cannot be supported by evidence-neither regarding the present COVID-19 pandemic, nor regarding the 1918-1920 Spanish Flu and other less-severe pandemics in the past. The price tag of lockdowns in terms of public health is high: by using the known connection between health and wealth, we estimate that lockdowns may claim 20 times more life years than they save. It is suggested therefore that a thorough cost-benefit analysis should be performed before imposing any lockdown for either COVID-19 or any future pandemic.


Calcitonin gene-related peptide analogues with aza and indolizidinone amino acid residues reveal conformational requirements for antagonist activity at the human calcitonin gene-related peptide 1 receptor.
Calcitonin gene-related peptide antagonists have potential for the treatment and prevention of disease states such as non-insulin-dependent diabetes mellitus, migraine headache, pain, and inflammation. To gain insight into the spatial requirements for CGRP antagonism, three strategies were employed to restrict the conformation of the potent undecapeptide antagonist, [D31,P34,F35]CGRP27-37. First, aza-amino acid scanning was performed, and ten aza-peptide analogues were synthesized and examined for biological activity. Second, (3S,6S,9S)-2-oxo-3-amino-indolizidin-2-one amino acid (I2aa) and (2S,6S,8S)-9-oxo-8-amino-indolizidin-9-one amino acid (I9aa) both were introduced at positions 31-32, 32-33, 33-34, and 34-35, regions of the backbone expected to adopt turns. Finally, the conformation of the backbone and side-chain of the C-terminal residue, Phe35-Ala36-Phe37-NH2, was explored employing (2S,4R,6R,8S)-9-oxo-8-amino-4-phenyl-indolizidin-9-one amino acid (4-Ph-I9aa) as a constrained phenylalanine mimic. The structure-activity relationships exhibited by our 26 analogues illustrate conformational requirements important for designing CGRP antagonists and highlight the importance of beta-turns centered at Gly33-Pro34 for potency.


Differential MMP-14 targeting by biglycan, decorin, fibromodulin, and lumican unraveled by in silico approach.
Small leucine-rich proteoglycans (SLRPs) are major regulators of extracellular matrix assembly and cell signaling. Lumican, a member of the SLRPs family, and its derived peptides were shown to possess antitumor activity by interacting directly with the catalytic domain of MMP-14 leading to the inhibition of its activity. The aim of the present report was to characterize by in silico three-dimensional (3D) modeling the structure and the dynamics of four SLRPs including their core protein and their specific polysaccharide chains to assess their capacity to bind to MMP-14 and to regulate its activity. Molecular docking experiments were performed to identify the specific amino acids of MMP-14 interacting with each of the four SLRPs. The inhibition of each SLRP (100 nM) on MMP-14 activity was measured and the constants of inhibition (Ki) were evaluated. The impact of the number of glycan chains, structures, and dynamics of lumican on the interaction with MMP-14 was assessed by molecular dynamics simulations. Molecular docking analysis showed that all SLRPs bind to MMP-14 through their concave face, but in different regions of the catalytic domain of MMP-14. Each SLRPs inhibited significantly the MMP-14 activity. Finally, molecular dynamics showed the role of glycan chains in interaction with MMP-14 and shielding effect of SLRPs. Altogether, the results demonstrated that each SLRP exhibited inhibition of MMP-14 activity. However, the differential targeting of MMP-14 by the SLRPs was shown to be related not only to the core protein conformation but also to the glycan chain structures and dynamics.


[Virtual reality in upper extremity dysfunction: specific features of usage in acute stroke].
Stroke is a leading cause of permanent disability. Being a new technology for neurorehabilitation, virtual reality (VR) allows for intensive trainings with a larger number of repetitions focused on mastering specific skills in patients with upper limb dysfunction. To date, there are insufficient studies evaluating the use of VR in the acute period of a stroke. To investigate the effectiveness and safety of adding VR trainings to standard therapy for improving upper limb function and for enhancing activities in daily life in patients with acute stroke. The investigation enrolled 78 patients with acute stroke (the median time from stroke onset, 3.7 days; median age, 63 (43; 79.3) years), who were randomized into 2 groups: a study group (standard therapy + VR) and a control one (standard therapy only). The patients of the study group underwent a VR training cycle lasting 15 minutes twice daily for 10 days. Before and after the training cycle, the British Medical Research Council scale for muscle strength, the upper extremity motor function sections of the Fugl-Meyer assessment (FMA) scale, hand dynamometry, and the nine hole peg test (NHPT) were used to evaluate upper extremity function in the participants; Sections 6 and 7 of the motor function assessment scale (MAS), the modified Rankin scale (MRS), and the total motor function scores of the functional independence measure (FIM) utilizing a 7-point ordinal scale were employed to assess everyday activity limitations. On completion of rehabilitation measures, the participants in the study and control groups showed a statistically significant improvement in upper extremity function and activities of daily living. According to the ratings of the FMA subscales 'hand' (p=0.034), 'hand (speed)' (p<0,001), and the total score of this scale (p=0.035) and according to those of the FIM scale (p=0.045), the patients of the study group demonstrated a statistically significant improvement compared with those in the control group. Assessments using the MRC scale, FMA subscales 'upper extremity' and 'wrist', NHPT, MAS, the Barthel index (BI), and MRS revealed no differences between the groups. No serious adverse events were recorded in the participants of the study group during the study period. The vast majority of studies evaluating the effect of VR in upper extremity dysfunction are conducted in patients with stroke lasting longer than 3 months. Despite the validity of using VR in the acute period of cerebral lesion, the investigations of this design are not numerous. At the same time, these studies often have limitations, such as a small number of participants (usually less than 10); a small number of training VR sessions; lack of 'blinding'; different types of software and hardware systems that implement VR technology; heterogeneity of participants according to the degree of upper extremity dysfunction; insufficient detailing of the content and scope of conventional therapy; comparison of VR with basic methods of motor rehabilitation, and sometimes the absence of a control group. Thus, the use of VR in addition to standard rehabilitation measures in the acute period of stroke favors improvements in upper extremity function and a reduction in daily limitations and is safe.


Repair of partial lesions with nerve grafting methods in the rat tibial nerve.
A partial nerve lesion and associated neuroma can be either left alone or repaired with a graft. A by-pass graft around the undisturbed lesion with end-to-side attachments might be a good alternative. This study in rats examines these strategies using walking-track analysis, muscle weights, and histology. After a tibial nerve partial lesion (3 mm) and a 21-day delay, the reexposed lesion was either not repaired, repaired with an interposed allograft, or a by-pass allograft. Functional results showed that all three groups had a steady improvement over the 8-week period, but without significant group differences. Gastrocnemius muscle ratios reflected intermediate atrophy. Axons regenerating through the lesion were more advanced than those which regenerated through the grafts and a neuroma was absent. The partial lesion can regenerate to an intermediate level without any intervention, including a by-pass graft, although the delayed repair strategy may have counteracted any potential benefits.


Activation and induction of antigen-specific T follicular helper cells play a critical role in recombinant SARS-CoV-2 RBD vaccine-induced humoral responses.
The role of follicular T helper (Tfh) cells in humoral response has been considered essential in recent years. Understanding how Tfh cells control complex humoral immunity is critical to developing strategies to improve the efficacy of vaccines against SARS-CoV-2 and other emerging pathogens. However, the immunologic mechanism of Tfh cells in SARS-CoV-2 receptor binding domain (RBD) vaccine strategy is limited. In this study, we expressed and purified recombinant SARS-CoV-2 RBD protein in Drosophila S2 cells for the first time and explored the mechanism of Tfh cells induced by RBD vaccine in humoral immune response. We mapped the dynamic of Tfh cell in lymph node and spleen following RBD vaccination and revealed the relationship between Tfh cells and humoral immune response induced by SARS-CoV-2 RBD vaccine through correlation analysis, blocking of IL-21 signaling pathway, and co-culture of Tfh with memory B cells. Recombinant RBD protein elicited a predominant Tfh1 and Tfh1-17 subset response and strong GC responses in spleen and lymph nodes, especially to enhanced vaccination. IL-21 secreted by Tfh cells affected the development and differentiation of B cells and played a key role in the humoral immune response. These observations will help us further understand the mechanism of protective immune response induced by COVID-19 vaccine and has guiding significance for the development of vaccines against newly emerging mutants.


Effect of Knockout of Mdr1a and Mdr1b ABCB1 Genes on the Systemic Exposure of a Doxorubicin-Conjugated Block Copolymer in Mice.
We previously elucidated that ATP-binding cassette subfamily B member 1 (ABCB1) mediates the efflux of doxorubicin-conjugated block copolymers from HeLa cells. Here, we investigated the role of ABCB1 in the in vivo behavior of a doxorubicin-conjugated polymer in Mdr1a/1b(-/-) mice. The area under the curve for intravenously administered polymer in Mdr1a/1b(-/-) mice was 2.2-fold greater than that in wild-type mice. The polymer was mostly distributed in the liver followed by spleen and less so in the brain, heart, kidney, and lung. The amount of polymer excreted in the urine was significantly decreased in Mdr1a/1b(-/-) mice. The amounts of polymers excreted in the feces were similar in both groups despite the higher systemic exposure in Mdr1a/1b(-/-) mice. Confocal microscopy images showed polymer localized in CD68(+) macrophages in the liver. These results show that knockout of ABCB1 prolonged systemic exposure of the doxorubicin-conjugated polymer in mice. Our results suggest that ABCB1 mediated the excretion of doxorubicin-conjugated polymer in urine and feces. Our results provide valuable information about the behavior of block copolymers in vivo, which is important for evaluating the pharmacokinetics of active substances conjugated to block copolymers or the accumulation of block copolymers in vivo.


Direct electron transfer of cytochrome P450 2B4 at electrodes modified with nonionic detergent and colloidal clay nanoparticles.
A method for construction of biosensors with membranous cytochrome P450 isoenzymes was developed based on clay/detergent/protein mixed films. Thin films of sodium montmorillonite colloid with incorporated cytochrome P450 2B4 (CYP2B4) with nonionic detergent were prepared on glassy carbon electrodes. The modified electrodes were electrochemically characterized, and bioelectrocatalytic reactions were followed. CYP2B4 can be reduced fast on clay-modified glassy carbon electrodes in the presence of the nonionic detergent Tween 80. In anaerobic solutions, reversible oxidation and reduction is obtained with a formal potential between -0.292 and -0.305 V vs Ag/AgCl 1 M KCl depending on the preparation of the biosensor. In air-saturated solution, bioelectrocatalytic reduction currents can be obtained with the CYP2B4-modified electrode on addition of typical substrates such as aminopyrine and benzphetamine. This reaction was suppressed when methyrapone, an inhibitor of P450 reactions, was present. Measurement of product formation also indicates the bioelectrocatalysis by CYP2B4.


The sialyltransferase ST3GAL6 influences homing and survival in multiple myeloma.
Glycosylation is a stepwise procedure of covalent attachment of oligosaccharide chains to proteins or lipids, and alterations in this process, especially increased sialylation, have been associated with malignant transformation and metastasis. The role of altered sialylation in multiple myeloma (MM) cell trafficking has not been previously investigated. In the present study we identified high expression of β-galactoside α-2,3-sialyltransferase, ST3GAL6, in MM cell lines and patients. This gene plays a key role in selectin ligand synthesis in humans through the generation of functional sialyl Lewis X. In MRC IX patients, high expression of this gene is associated with inferior overall survival. In this study we demonstrate that knockdown of ST3GAL6 results in a significant reduction in levels of α-2,3-linked sialic acid on the surface of MM cells with an associated significant reduction in adhesion to MM bone marrow stromal cells and fibronectin along with reduced transendothelial migration in vitro. In support of our in vitro findings, we demonstrate significantly reduced homing and engraftment of ST3GAL6 knockdown MM cells to the bone marrow niche in vivo, along with decreased tumor burden and prolonged survival. This study points to the importance of altered glycosylation, particularly sialylation, in MM cell adhesion and migration.


Military surgery in the 21st century.
The battlefield has changed. UK and US Forces in Iraq and Afghanistan are engaged in asymmetric, three bloc warfare. Three bloc warfare indicates that what is effectively war-fighting occurs in parallel with peace-keeping and humanitarian operations. Casualties are numerically lower than in previous conflicts but many survivors are horrifically mutilated as body armour prevents the fatal injury in this severely injured group. Rapid evacuation of severely injured from the point of injury, usually by helicopter, provides extreme challenges for surgical teams. Damage control surgery requires support in the form of environmental control, adequate diagnostics (including computerised tomography) and effective intensive care facilities if the patient is to survive. Teams need to be highly trained and to have experience of complex surgery and trauma. They must be adequately resourced if lives are to be saved.


And young child feeding practices in different country settings.
Alive & Thrive aims to increase exclusive breastfeeding and complementary feeding practices in Bangladesh, Ethiopia, and Vietnam. To develop and execute comprehensive communication strategies adapted to each context. We documented how three countries followed an established iterative planning process, with research steps followed by key decisions, to develop a communication strategy in each country. Secondary analysis and formative research identified the priority practices to focus on, and locally specific constraints to proper infant and young child feeding (IYCF). Communication strategies were then developed based on the social, cultural, economic, epidemiological, media use, and programmatic contexts of each country. There were widespread gaps between recommended and actual feeding practices, and these varied by country. Gaps were identified in household, community, and institutional levels of awareness and skills. Strategies were designed that would enable mothers in each specific setting to adopt practices. To improve priority behaviors, messaging and media strategies addressed the most salient behavioral determinants through face-to-face communication, social mobilization, and mass media. Trials of improved practices (TIPs), concept testing, and pretesting of materials proved useful to verify the relevance and likely effectiveness of communication messages and materials tailored for different audiences in each setting. Coordination and collaboration with multiple stakeholders from the start was important to harmonize messages and approaches, expand geographic coverage to national scale, and sustain the interventions. Our experience with designing large-scale communication strategies for behavior change confirms that systematic analysis and local planning cannot be omitted from the critical process of strategic design tailored to each context. Multiple communication channels matched to media habits in each setting can reach a substantial proportion of mothers and others who influence their IYCF practices. Preliminary data suggest that exposure to mass media plays a critical role in rapidly reaching mothers, household members, community influentials, and health workers on a large scale. Combining face-to-face interventions for mothers with social mobilization and mass media was effective in improving IYCF practices.


The Supportive Care Needs of Regional and Remote Cancer Caregivers.
As cancer survival rates continue to increase, so will the demand for care from family and friends, particularly in more isolated settings. This study aims to examine the needs of cancer caregivers in regional and remote Australia. A total of 239 informal (i.e., non-professional) cancer caregivers (e.g., family/friends) from regional and remote Queensland, Australia, completed the Comprehensive Needs Assessment Tool for Cancer Caregivers (CNAT-C). The frequencies of individuals reporting specific needs were calculated. Logistic regression analyses assessed the association between unmet needs and demographic characteristics and cancer type. The most frequently endorsed needs were lodging near hospital (77%), information about the disease (74%), and tests and treatment (74%). The most frequent unmet needs were treatment near home (37%), help with economic burden (32%), and concerns about the person being cared for (32%). Younger and female caregivers were significantly more likely to report unmet needs overall (OR = 2.12; OR = 0.58), and unmet healthcare staff needs (OR = 0.35; OR = 1.99, respectively). Unmet family and social support needs were also significantly more likely among younger caregivers (OR = 0.35). Caregivers of breast cancer patients (OR = 0.43) and older caregivers (OR = 0.53) were significantly less likely to report unmet health and psychology needs. Proportions of participants reporting needs were largely similar across demographic groups and cancer type with some exceptions. Caregiver health, practical issues associated with travel, and emotional strain are all areas where regional and remote caregivers require more support. Caregivers' age and gender, time since diagnosis and patient cancer type should be considered when determining the most appropriate supportive care.


A survey of aquarium fishes, their foods, and environmental water for salmonellae.
The presence of salmonellae in the pet shop environment, fishes, aquarium water, and in a variety of commercially prepared and natural fish foods was determined. Salmonellosis attributed to turtles purchased from pet shops has been responsible for an estimated 280,000 human cases annually in the United States. The possibility that aquarium fishes, or their food and aquatic habitat might also serve as sources of human salmonellosis was deemed worthy of clarification. Results of the present survey indicate that this is probably not correct, and a public health hazarddoes not exist.


"What Is Right for Me, Is Not Necessarily Right for You": The Endogenous Factors Influencing Nonparticipation in Medical Assistance in Dying.
Access to medical assistance in dying (MAID) is influenced by legislation, health care providers (HCPs), the number of patient requests, and the patients' locations. This research explored the factors that influenced HCPs' nonparticipation in formal MAID processes and their needs to support this emerging practice area. Using an interpretive description methodology, we interviewed 17 physicians and 18 nurse practitioners who identified as non-participators in formal MAID processes. Nonparticipation was influenced by their (a) previous personal and professional experiences, (b) comfort with death, (c) conceptualization of duty, (d) preferred end-of-life care approaches, (e) faith or spirituality beliefs, (f) self-accountability, (g) consideration of emotional labor, and (h) future emotional impact. They identified a need for clear care pathways and safe passage. Two separate yet overlapping concepts were identified, conscientious objection to and nonparticipation in MAID, and we discussed options to support the social contract of care between HCPs and patients.


Word frequency cues word order in adults: cross-linguistic evidence.
One universal feature of human languages is the division between grammatical functors and content words. From a learnability point of view, functors might provide entry points or anchors into the syntactic structure of utterances due to their high frequency. Despite its potentially universal scope, this hypothesis has not yet been tested on typologically different languages and on populations of different ages. Here we report a corpus study and an artificial grammar learning experiment testing the anchoring hypothesis in Basque, Japanese, French, and Italian adults. We show that adults are sensitive to the distribution of functors in their native language and use them when learning new linguistic material. However, compared to infants' performance on a similar task, adults exhibit a slightly different behavior, matching the frequency distributions of their native language more closely than infants do. This finding bears on the issue of the continuity of language learning mechanisms.


An integrated network analysis, RNA-seq and in vivo validation approaches to explore the protective mechanism of Mongolian medicine formulae Ruda-6 against indomethacin-induced gastric ulcer in rats.
Gastric ulcer (GU) is one of the most prevalent digestive diseases that seriously affects people's health. Previous studies have demonstrated the anti-GU effect of Ruda-6 (RD-6), a classic formulae of traditional Mongolian medicine. However, the underlying mechanism of RD-6 against GU remains elusive. Thus, we conducted an integrative approach of network analysis, RNA-seq, and in vivo validation experiment to elucidate the therapeutic mechanisms of RD-6 in preventing GU. A network analysis was performed to predict the potential targets of RD-6. Rats were pretreated with RD-6 at different doses for 21 days, followed by GU induction with indomethacin injection. The ulcer index and inhibition rates were calculated, and the levels of inflammatory related factors were determined by ELISA. The gastroprotective mechanism of RD-6 against ulceration was verified by RNA-seq and the key pathway was detected by in vivo validation. As the network analysis predicted, RD-6 exerts anti-GU effects by regulating 75 targets and 160 signaling pathways. Animal experiment results suggested that pretreatment with RD-6 significantly ameliorated the gastric mucosal injury and inflammatory response, as evidenced by a reduced ulcer index, decreased interleukin (IL)-1β, IL-6, and IL-17 levels, and increased prostaglandin E2 (PGE2) levels in the GU model rats induced by indomethacin. RNA-seq data identified four potential hub genes that were primarily involved in the IL-17 signaling pathway. Furthermore, in vivo validation experiment showed that RD-6 inhibited the IL-17 signaling pathway by down-regulating the expression of IL17RA, proto-oncogene C-Fos (FOS), IL1B and prostaglandin-endoperoxide synthase 2 (PTGS2). Taken together, the present study provides evidence that RD-6 could effectively protect against indomethacin-induced GU, which might be attributed to suppressed inflammation. The IL-17 signaling pathway may be one of the crucial mechanisms that mediates the effect of RD-6.


Psychological consequences of cystic fibrosis in adults.
Increasing numbers of cystic fibrosis sufferers are surviving to adulthood. This article reviews the findings of psychological research into this group during the last decade. The psychological difficulties met by patients, families and doctors are reported. Illness coping strategies developed by sufferers, which may be applicable to other chronic diseases, are discussed.


Paroxysmal hypertension, pheochromocytoma, and pregnancy.
Hypertension is the most common medical complication of pregnancy. Pheochromocytoma in pregnancy is rare, and if unrecognized, can cause serious perinatal morbidity and mortality. A patient with severe hypertension, postpartum pulmonary edema, and a recognized pheochromocytoma is described. Abdominal palpation after vaginal childbirth reproduced the diagnostic triad of hypertension, headaches, and palpitations. Magnetic resonance imaging established the correct diagnosis before biochemical confirmation of excess catecholamine production. The patient responded to alpha-adrenergic receptor blockade with control of her severe hypertension and clearing of pulmonary edema. The best time to diagnose a pheochromocytoma is before delivery because vaginal childbirth stimulates the release of lethal amounts of catecholamines. The physician who delivers babies must distinguish between labile hypertension and paroxysmal hypertension. Most experts believe that a spontaneous vaginal delivery is contraindicated when the patient has a pheochromocytoma. Postpartum pulmonary edema associated with a pheochromocytoma is unusual. The profound pressor response elicited by palpation of the postpartum abdomen, the failure of medications usually effective in the treatment of a hypertensive crisis, and the use of magnetic resonance imaging to confirm a functioning adrenal adenoma are the features unique to this case.


The "Cheshire Cat" escape strategy of the coccolithophore Emiliania huxleyi in response to viral infection.
The coccolithophore Emiliania huxleyi is one of the most successful eukaryotes in modern oceans. The two phases in its haplodiploid life cycle exhibit radically different phenotypes. The diploid calcified phase forms extensive blooms, which profoundly impact global biogeochemical equilibria. By contrast, the ecological role of the noncalcified haploid phase has been completely overlooked. Giant phycodnaviruses (Emiliania huxleyi viruses, EhVs) have been shown to infect and lyse diploid-phase cells and to be heavily implicated in the regulation of populations and the termination of blooms. Here, we demonstrate that the haploid phase of E. huxleyi is unrecognizable and therefore resistant to EhVs that kill the diploid phase. We further show that exposure of diploid E. huxleyi to EhVs induces transition to the haploid phase. Thus we have clearly demonstrated a drastic difference in viral susceptibility between life cycle stages with different ploidy levels in a unicellular eukaryote. Resistance of the haploid phase of E. huxleyi provides an escape mechanism that involves separation of meiosis from sexual fusion in time, thus ensuring that genes of dominant diploid clones are passed on to the next generation in a virus-free environment. These "Cheshire Cat" ecological dynamics release host evolution from pathogen pressure and thus can be seen as an opposite force to a classic "Red Queen" coevolutionary arms race. In E. huxleyi, this phenomenon can account for the fact that the selective balance is tilted toward the boom-and-bust scenario of optimization of both growth rates of calcifying E. huxleyi cells and infectivity of EhVs.


Characterization of the laser-induced blue, green and red fluorescence signatures of leaves of wheat and soybean grown under different irradiance.
The blue, green and red fluorescence emission of green wheat (Triticum aestivum L. var. Rector) and soybean leaves (Glycine max L. var. Maple Arrow) as induced by UV light (nitrogen laser: 337 nm) was determined in a phytochamber and in plants grown in the field. The fluorescence emission spectra show a blue maximum near 450 nm, a green shoulder near 530 nm and the two red chlorophyll fluorescence maxima near 690 and 735 nm. The ratio of blue to red fluorescence, F450/F690, exhibited a clear correlation to the irradiance applied during the growth of the plants. In contrast, the chlorophyll fluorescence ratio, F690/F735, and the ratio of blue to green fluorescence, F450/F530, seem not to be or are only slightly influenced by the irradiance applied during plant growth. The blue fluorescence F450 only slightly decreased, whereas the red chlorophyll fluorescence decreased with increasing irradiance applied during growth of the plants. This, in turn, resulted in greatly increased values of the ratio, F450/F690, from 0.5 - 1.5 to 6.4 - 8.0. The decrease in the chlorophyll fluorescence with increasing irradiance seems to be caused by the accumulation of UV light absorbing substances in the epidermal layer which considerably reduces the UV laser light which passes through the epidermis and excites the chlorophyll fluorescence of the chloroplasts in the subepidermal mesophyll cells.


[Neuropsychological approach to the complex study of senile dementia].
Thirty-eight patients with late mentally deteriorating processes were studied. Using a standardized neuropsychological method, the authors have outlined four major variants of neuropsychological syndromes characteristic of damage to definite zones of the brain. Comparison of these data with the results of clinical and computer-aided tomographic examination showed that the neuropsychological approach is an adequate method in the combined study of senile dementia.


Depression, coping and level of neurological impairment in multiple sclerosis.
This study examined the relationship between coping and depression in multiple sclerosis patients, and how that relationship varies at different levels of physical impairment. One-hundred and one patients with clinically definite MS were assessed using the Kurtzke Expanded Disability Status Scale (EDSS), the Ways of Coping Inventory (WCI) with three sub-scales developed by Wineman et al, and the Beck Depression Inventory (BDI). Depression was significantly higher at more advanced levels of neurologic impairment than at lower levels. Escape-Avoidance and Emotional Respite were positively related to level of depression. Planful Problem-Solving and Cognitive Reframing were negatively related to depression. An interaction between coping, depression, and level of neurologic impairment was observed in which Planful Problem-Solving and Cognitive Reframing were more strongly related to depression at higher levels of impairment. The interaction effect for Escape-Avoidance and Emotional Respite with depression and level of impairment did not reach significance. It was concluded that there is a significant interaction between level of neurologic impairment, coping behaviors, and depression in patients with MS.


Opening Minds Stigma Scale for Health Care Providers (OMS-HC): examination of psychometric properties and responsiveness.
Diminishing stigmatization for those with mental illnesses by health care providers (HCPs) is becoming a priority for programming and policy, as well as research. In order to be successful, we must accurately measure stigmatizing attitudes and behaviours among HCPs. The Opening Minds Stigma Scale for Health Care Providers (OMS-HC) was developed to measure stigma in HCP populations. In this study we revisit the factor structure and the responsiveness of the OMS-HC in a larger, more representative sample of HCPs that are more likely to be targets for anti-stigma interventions. Baseline data were collected from HCPs (n = 1,523) during 12 different anti-stigma interventions across Canada. The majority of HCPs were women (77.4%) and were either physicians (MDs) (41.5%), nurses (17.0%), medical students (13.4%), or students in allied health programs (14.0%). Exploratory factor analysis (EFA) was conducted using complete pre-test (n = 1,305) survey data and responsiveness to change analyses was examined with pre and post matched data (n = 803). The internal consistency of the OMS-HC scale and subscales was evaluated using the Cronbach's alpha coefficient. The scale's sensitivity to change was examined using paired t-tests, effect sizes (Cohen's d), and standardized response means (SRM). The EFA favored a 3-factor structure which accounted for 45.3% of the variance using 15 of 20 items. The overall internal consistency for the 15-item scale (α = 0.79) and three subscales (α = 0.67 to 0.68) was acceptable. Subgroup analysis showed the internal consistency was satisfactory across HCP groups including physicians and nurses (α = 0.66 to 0.78). Evidence for the scale's responsiveness to change occurred across multiple samples, including student-targeted interventions and workshops for practicing HCPs. The Social Distance subscale had the weakest level of responsiveness (SRM ≤ 0.50) whereas the more attitudinal-based items comprising the Attitude (SRM ≤ 0.91) and Disclosure and Help-seeking (SRM ≤ 0.68) subscales had stronger responsiveness. The OMS-HC has shown to have acceptable internal consistency and has been successful in detecting positive changes in various anti-stigma interventions. Our results support the use of a 15-item scale, with the calculation of three sub scores for Attitude, Disclosure and Help-seeking, and Social Distance.


Determination of phthalic acid diesters in human milk at low ppb levels.
Phthalic acid diesters (PAE) are omnipresent in the human environment and food is a major contributor to the overall human exposure towards these chemicals. Due to developmental effects, PAE infants' exposure via human milk has been subjected to a number of analytical studies. These previous studies, however, revealed that normal laboratory blank values are in the range of or even higher than human milk levels due to the presence of PAE in laboratory environments. In order to provide more reliable data on PAE exposure via human milk, the aim of this study was to develop and validate a robust and sensitive analytical method. This should be capable of removing matrix components efficiently and guarantee limits of quantification in the low ppb range. The method development took into account liquid-liquid extraction and selective pressurised fluid extraction (sPFE) as well as chromatography-based clean-up steps. The final method consisted of a liquid-liquid extraction followed by an automated chromatographic clean-up by an sPFE device. After volume reduction the cleaned extracts were analysed by quadrupole GC/MS. Quantification was based on internal standards. An extensive quality assurance and method test programme demonstrated conservatively determined limits of detection and quantification from 0.3 to 10 ng g⁻¹ in human milk, with recoveries of internal standards from 50% to 101%. Thus, the method allowed the quality-assured detection of di-isobutyl phthalate (DiBP), di-n-butyl phthalate (DBP), di-2-ethylhexyl phthalate (DEHP), di-allyl phthalate (DAP), benzylbutyl phthalate (BBP) and di-cyclohexyl phthalate (DcHP) in 30 human milk samples provided by 30 volunteers from southern Germany. DiBP, DBP and DEHP were the most commonly detected PAE, with median levels of 1.0, 0.6 and 2.3 ng g⁻¹, respectively.


Assessment of strategies for switching patients from olanzapine to risperidone: a randomized, open-label, rater-blinded study.
In clinical practice, physicians often need to change the antipsychotic medications they give to patients because of an inadequate response or the presence of unacceptable or unsafe side effects. However, there is a lack of consensus in the field as to the optimal switching strategy for antipsychotics, especially with regards to the speed at which the dose of the previous antipsychotic should be reduced. This paper assesses the short-term results of strategies for the discontinuation of olanzapine when initiating risperidone. In a 6-week, randomized, open-label, rater-blinded study, patients with schizophrenia or schizoaffective disorder, on a stable drug dose for more than 30 days at entry, who were intolerant of or exhibiting a suboptimal symptom response to more than 30 days of olanzapine treatment, were randomly assigned to the following switch strategies (common risperidone initiation scheme; varying olanzapine discontinuation): (i) abrupt strategy, where olanzapine was discontinued at risperidone initiation; (ii) gradual 1 strategy, where olanzapine was given at 50% entry dose for 1 week after risperidone initiation and then discontinued; or (iii) gradual 2 strategy, where olanzapine was given at 100% entry dose for 1 week, then at 50% in the second week, and then discontinued. The study enrolled 123 patients on stable doses of olanzapine. Their mean age was 40.3 years and mean (+/- standard deviation (SD)) baseline Positive and Negative Syndrome Scale (PANSS) total score of 75.6 +/- 11.5. All-cause treatment discontinuation was lowest (12%) in the group with the slowest olanzapine dose reduction (gradual 2) and occurred at half the discontinuation rate in the other two groups (25% in abrupt and 28% in gradual 1). The relative risk of early discontinuation was 0.77 (confidence interval 0.61-0.99) for the slowest dose reduction compared with the other two strategies. After the medication was changed, improvements at endpoint were seen in PANSS total score (-7.3; p < 0.0001) and in PANSS positive (-3.0; p < 0.0001), negative (-0.9; p = 0.171) and anxiety/depression (-1.4; p = 0.0005) subscale scores. Severity of movement disorders and weight changes were minimal. When switching patients from olanzapine to risperidone, a gradual reduction in the dose of olanzapine over 2 weeks was associated with higher rates of retention compared with abrupt or less gradual discontinuation. Switching via any strategy was associated with significant improvements in positive and anxiety symptoms and was generally well tolerated. ClinicalTrials.gov NCT00378183.


Integral role of PTP1B in adiponectin-mediated inhibition of oncogenic actions of leptin in breast carcinogenesis.
The molecular effects of obesity are mediated by alterations in the levels of adipocytokines. High leptin level associated with obese state is a major cause of breast cancer progression and metastasis, whereas adiponectin is considered a "guardian angel adipocytokine" for its protective role against various obesity-related pathogenesis including breast cancer. In the present study, investigating the role of adiponectin as a potential inhibitor of leptin, we show that adiponectin treatment inhibits leptin-induced clonogenicity and anchorage-independent growth. Leptin-stimulated migration and invasion of breast cancer cells is also effectively inhibited by adiponectin. Analyses of the underlying molecular mechanisms reveal that adiponectin suppresses activation of two canonical signaling molecules of leptin signaling axis: extracellular signal-regulated kinase (ERK) and Akt. Pretreatment of breast cancer cells with adiponectin protects against leptin-induced activation of ERK and Akt. Adiponectin increases expression and activity of the physiological inhibitor of leptin signaling, protein tyrosine phosphatase 1B (PTP1B), which is found to be integral to leptin-antagonist function of adiponectin. Inhibition of PTP1B blocks adiponectin-mediated inhibition of leptin-induced breast cancer growth. Our in vivo studies show that adenovirus-mediated adiponectin treatment substantially reduces leptin-induced mammary tumorigenesis in nude mice. Exploring therapeutic strategies, we demonstrate that treatment of breast cancer cells with rosiglitazone results in increased adiponectin expression and inhibition of migration and invasion. Rosiglitazone treatment also inhibits leptin-induced growth of breast cancer cells. Taken together, these data show that adiponectin treatment can inhibit the oncogenic actions of leptin through blocking its downstream signaling molecules and raising adiponectin levels could be a rational therapeutic strategy for breast carcinoma in obese patients with high leptin levels.


An fMRI investigation of the role of the basal ganglia in reasoning.
Neuropsychological studies highlight the importance of the prefrontal cortex in abstract reasoning ability. The prefrontal cortex is anatomically connected to the basal ganglia through a series of parallel loops. The dorsolateral prefrontal cortex shares relatively greater connectivity with the caudate head, and it has been proposed that the caudate head may be more sensitive to executive processing. To investigate the frontostriatal circuitry underlying abstract reasoning, we designed a reasoning task in which stimuli varied sequentially across one dimension. Critically, the task required that subjects deduce and apply a sequencing rule. We also developed a match-to-sample working memory task in order to identify and contrast brain regions involved in working memory. We observed reasoning-related activation in both cortical and subcortical regions. Subcortically, the caudate was found to be bilaterally active during both the reasoning and matching tasks, and the left caudate head was more active for reasoning compared to matching. An anatomically defined region of interest analysis demonstrated activity for both reasoning and matching tasks within the body of the caudate. In contrast, the left caudate head was found to more specifically support reasoning. Cortically, we observed activity of bilateral rostrolateral prefrontal cortex, right ventrolateral prefrontal cortex, and bilateral parietal cortex during reasoning. Our results suggest that reasoning requires an interaction between cortical areas and the caudate nucleus, in which the caudate body supports both reasoning and working memory and the head of the caudate is specifically involved in the executive demands of reasoning, namely deducing and applying a sequence rule.


Prognostic and Predictive Value of a 15 Transcription Factors (TFs) Panel for Hepatocellular Carcinoma.
Hepatocellular carcinoma (HCC) is one of the most devastating diseases worldwide. Limited performance of clinicopathologic parameters as prognostic factors underscores more accurate and effective biomarkers for high-confidence prognosis that guide decision-making for optimal treatment of HCC. The aim of the present study was to establish a novel panel to improve prognosis prediction of HCC patients, with a particular interest in transcription factors (TFs). A TF-related prognosis model of liver cancer with data from ICGC-LIRP-JI cohort successively were processed by univariate and multivariate Cox regression analysis. Then, for evaluating the prognostic prediction value of the model, receiver operating characteristic (ROC) curve and survival analysis were performed both with internal data from the International Cancer Genome Consortium (ICGC) and external data from The Cancer Genome Atlas (TCGA). Furthermore, we verified the expression of three genes in HCC cell lines by Western blot and qPCR and protein expression level by IHC in liver cancer patients' sample. Finally, we constructed a TF clinical characteristics nomogram to furtherly predict liver cancer patient survival probability with TCGA cohort. By Cox regression analysis, a panel of 15 TFs (ZNF331, MYCN, AHRR, LEF1, ZNF780A, POU1F1, DLX5, ZNF775, PLSCR1, FOXK1, TAL2, ZNF558, SOX9, TCFL5, GSC) was identified to present with powerful predictive performance for overall survival of HCC patients based on internal ICGC cohort and external TCGA cohort. A nomogram that integrates these factors was established, allowing efficient prediction of survival probabilities and displaying higher clinical utility. The 15-TF panel is an independent prognostic factor for HCC, and 15 TF-based nomogram might provide implication an effective approach for HCC patient management and treatment.


The impact of postoperative complications on oncological outcomes of liver transplantation for hepatocellular carcinoma: A competing risk analysis.
To investigate the influence of postoperative complications on tumor-related (TRD), disease-free survival (DFS) and overall survival (OS) in patients undergoing liver transplant (LT) for hepatocellular carcinoma (HCC). We retrospectively evaluated 425 LTs for HCC from 2010 to 2019. Postoperative complications were classified according to Comprehensive Complication Index (CCI) and the posttransplant risk of TRD assessed through Metroticket 2.0 calculator. The population was stratified into high-risk and low-risk cohorts based on the predicted TRD risk of 80%. In a second step, we re-evaluated TRD, DFS and OS of both cohorts according to a further stratification based on 47.3 points of CCI cut-off. In the low-risk cohort, we observed a significantly better DFS (84% vs. 46%, p<0.001), TRD (3% vs. 26%, p<0.001) and OS (89% vs. 62%, p<0.001) in the group with CCI < 47.3. In the high-risk cohort, patients with CCI < 47.3 had significantly better DFS (50% vs. 23%, p = 0.003) and OS (68% vs. 42%, p = 0.02) and a comparable TRD (22% vs. 31%, p = 0.142). A complicated postoperative course negatively influenced long-term survival. This poorer oncological outcome associated with in-hospital postoperative complications suggests that every effort should be made to improve the early posttransplant course in HCC patients, including a careful donor-to recipient match and use of new perfusion technologies.


Bone surrogates provide authentic onlay graft fixation torques.
Onlay graft bone augmentation is a standard practice to restore the loss of height of the alveolar ridge following loss of a tooth. Cranial grafts, lifted from the parietal bone, are sandwiched and used to bridge the bony defect in the jaw by means of small screws. During the elevation of the covering gum and subsequent screw placement, care has to be taken in order to preserve underlying nerves. Therefore, to avoid harm to the patient, a solid education of surgeons is essential, which requires training and experience. A simulator for cranial graft-lift training was already developed and shall be expanded to train the augmentation of the lifted implants. Therefore, in this study, synthetic bones for onlay block graft screw placement with realistic haptics for the screw application training were evaluated and compared with human specimens. Six different polyurethane based bone surrogate composites, enriched with varying amounts of calcium-based mineral fillers and blowing agents, were developed. The haptical properties of these synthetic bones were validated for screw placement and compared with human parietal bone specimens. For that, bones were pre-drilled, screws were automatically inserted using a customized testbench and the slope of the screw-insertion torques were analyzed. The slope of the screw insertion torques of the human reference bones was 56.5 ± 14.0 * 10-3 Nm/deg, Surrogates with lower amounts of mineral fillers and blowing agents showed lower torques than the human bone. Synthetic bones, validated for drilling, milling and sawing in an earlier study, also achieved significantly lower torques, which were only the half of the human parietal bones. Two intermediate stages of the aforementioned material compositions, consisting of 75% mineral filler with 0.75% blowing agent and 100% mineral filler with 1.00% blowing agent revealed results comparable with human bone (57.4 ± 10.2 *10-3 Nm/deg, p = 0.893 and 54.9 ± 11.1 *10-3 Nm/deg, p = 0.795, respectively). In conclusion, our findings suggest that, two newly developed polyurethane-based materials mimicking the haptical properties of an onlay bone graft screw fixation, have been identified. Thus, these surrogates are capable of mimicking real bone tissue in our simulator for the education of novice surgeons.


Comparative membrane proteomics analyses of breast cancer cell lines to understand the molecular mechanism of breast cancer brain metastasis.
Breast cancer is the leading type of cancer in women. Breast cancer brain metastasis is currently considered an issue of concern among breast cancer patients. Membrane proteins play important roles in breast cancer brain metastasis, involving cell adhesion and penetration of blood-brain barrier. To understand the mechanism of breast cancer brain metastasis, liquid chromatography-tandem mass spectrometry (LC-MS/MS) was employed in conjunction with enrichment of membrane proteins to analyze the proteomes from five different breast cancer and a brain cancer cell lines. Quantitative proteomic data of all cell lines were compared with MDA-MB-231BR which is a brain seeking breast cancer cell line, thus representing brain metastasis characteristics. Label-free proteomics of the six cell lines facilitates the identification of 1238 proteins and the quantification of 899 proteins of which more than 70% were membrane proteins. Unsupervised principal component analysis (PCA) of the label-free proteomics data resulted in a distinct clustering of cell lines, suggesting quantitative differences in the expression of several proteins among the different cell lines. Unique protein expressions in 231BR were observed for 28 proteins. The up-regulation of STAU1, AT1B3, NPM1, hnRNP Q, and hnRNP K and the down-regulation of TUBB4B and TUBB5 were noted in 231BR relative to 231 (precursor cell lines from which 231BR is derived). These proteins might contribute to the breast cancer brain metastasis. Ingenuity pathway analysis (IPA) supported the great brain metastatic propensity of 231BR and suggested the importance of the up-regulation of integrin proteins and down-regulation of EPHA2 in brain metastasis.


Cardiovascular Risk Factors Among Patients With Vestibular Neuritis.
To investigate the correlation between cardiovascular risk factors (CVRFs) and vestibular neuritis (VN) in hospitalized adult patients. A cross-sectional retrospective study was conducted in a tertiary hospital setting. The medical records of patients (aged over 18 years old) who were hospitalized between the years 2005 and 2014 with the diagnosis of VN were retrieved. Inclusion criteria were: (1) acute vertigo lasting for at least 24 hours, (2) absence of auditory complaints, (3) horizontal unidirectional nystagmus present during physical examination, and (4) absence of neurological symptoms or signs. The ratio of CVRFs among VN patients was compared to the ratio of those among the general Israeli population. A significantly higher prevalence of CVRFs was found among VN hospitalized patients in comparison to the general population ( P < .05). Furthermore, a significant correlation ( P < .001) was found between the patients' age and the number of CVRFs (r = .387). A positive correlation (r = .643) was found between the number of CVRFs and VN in each age group ( P = .119). There may be a possible interrelation between CVRFs and VN. This correlation can be caused by occlusion of small blood vessels leading to labyrinthine ischemia and apparition of symptoms of VN.


Toward Best Practice in Evaluation: A Study of Australian Health Promotion Agencies.
Evaluation makes a critical contribution to the evidence base for health promotion programs and policy. Because there has been limited research about the characteristics and determinants of evaluation practice in this field, this study audited evaluations completed by health promotion agencies in Victoria, Australia, and explored the factors that enabled or hindered evaluation performance. Twenty-four agencies participated. A systematic assessment of 29 recent evaluation reports was undertaken, and in-depth interviews were carried out with 18 experienced practitioners. There was wide variability in the scope of evaluations and the level of reporting undertaken. Formative evaluation was uncommon, but almost all included process evaluation, especially of strategy reach and delivery. Impact evaluation was attempted in the majority of cases, but the designs and measures used were often not specified. Practitioners strongly endorsed the importance of evaluation, but the reporting requirements and inconsistent administrative procedures of the funding body were cited as significant barriers. Budget constraints, employment of untrained coworkers, and lack of access to measurement tools were other major barriers to evaluation. Capacity building to strengthen evaluation needs to encompass system, organizational, and practitioner-level action. This includes strengthening funding and reporting arrangements, fostering partnerships, and tailoring workforce development opportunities for practitioners.


Application of the Target Lipid Model to Assess Toxicity of Heterocyclic Aromatic Compounds to Aquatic Organisms.
Heterocyclic aromatic compounds can be found in crude oil and coal and often co-exist in environmental samples with their homocyclic aromatic counterparts. The target lipid model (TLM) is a modeling framework that relates aquatic toxicity to the octanol-water partition coefficient (KOW ) that has been calibrated and validated for hydrocarbons. A systematic analysis of the applicability of the TLM to heterocyclic aromatic compounds has not been performed. The objective of the present study was to compile reliable toxicity data for heterocycles and determine whether observed toxicity could be successfully described by the TLM. Results indicated that the TLM could be applied to this compound class by adopting an empirically derived coefficient that accounts for partitioning between water and lipid. This coefficient was larger than previously reported for aromatic hydrocarbons, indicating that these heterocyclic compounds exhibit higher affinity to target lipid and toxicity. A mechanistic evaluation confirmed that the hydrogen bonding accepting moieties of the heteroatoms helped explain differences in partitioning behavior. Given the TLM chemical class coefficient reported in the present study, heterocyclic aromatics can now be explicitly incorporated in TLM-based risk assessments of petroleum substances, other products, or environmental media containing these compounds. Environ Toxicol Chem 2021;40:3000-3009. © 2021 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.


Biomechanical and energetic effects of a stance-control orthotic knee joint.
Users of traditional knee-ankle-foot orthoses (KAFOs) walk with either locked or unlocked knee joints depending on the level of stability required. Some users may benefit from new stance-control KAFOs that prevent stance-phase knee flexion but allow swing-phase flexion. We collected data from nine nondisabled adults who walked with KAFOs that incorporated the Horton Stance-Control Orthotic Knee Joint (SCOKJ) in the locked, unlocked, and auto (which provides knee stability during stance phase and knee flexion during swing phase) modes to investigate the biomechanical and energetic effects of stance-control orthoses. Studying nondisabled subjects allowed us to analyze the effects of stance-control orthoses in a homogenous population. In general, gait kinematics for the auto and unlocked modes were more similar than for the auto and locked modes. Despite the elimination of hip hiking in the auto mode, oxygen cost was not different between the auto and locked modes (p > 0.99). The SCOKJ allowed our nondisabled subjects to walk with a more normal gait pattern; however, future research should explore the effect of stance-control orthoses on persons with gait pathology.


Comparative drug use and psychosocial profiles of opioid dependents applying for medication versus medication-free treatment.
Drug treatment-seeking abusers generally self-select their treatment modality. Most opioid-dependent individuals select either methadone maintenance or detoxification followed by medication-free outpatient treatment. This study examined demographic, drug use, and drug abuse treatment history for opioid dependent patients entering an opioid agonist (n = 203) or a medication-free (n = 155) treatment program located on the same hospital campus. Similar demographic profiles support the assertion that samples were drawn from the same inner city urban opioid using population. Patients entering drug-free treatment, however, reported more extensive prior use of both cocaine and alcohol. Further, these patients had a generally more severe profile of psychosocial problems in medical, legal, employment, family/social, and psychiatric areas as measured by the Addiction Severity Index (ASI). These findings suggest that outpatient drug-free programs enrolling opioid abusers are faced with a unique service delivery challenge that involves service needs in multiple arenas. The findings further suggest that patient self-selection of treatment programs may in some cases result in a discrepancy between patient needs and the ability of treatment programs to retain patients and deliver an appropriate array of services.


Gutter Characteristics and Stent Compression of Self-Expanding vs Balloon-Expandable Chimney Grafts in Juxtarenal Aneurysm Models.
Purpose: To assess in silicone juxtarenal aneurysm models the gutter characteristics and compression of different types of chimney graft (CG) configurations. Materials and Methods: Fifty-seven combinations of Excluder C3 or Conformable Excluder stent-grafts (23, 26, and 28.5 mm) were deployed in 2 silicone juxtarenal aneurysm models with 3 types of CGs: Viabahn self-expanding (VSE; 6 and 13 mm) or Viabahn balloon-expandable (VBX; 6, 10, and 12 mm) stent-grafts and Advanta V12 balloon-expandable stent-grafts (ABX; 6 and 12 mm). Setups were divided into 4 groups on the basis of increasing CG and main graft (MG) diameters. Two independent observers assessed gutter size and type as well as CG compression on computed tomography scans using postprocessing software. Results: In the smaller diameter combinations (6-mm CG and 23-, 26-, and 28.5-mm MGs), both VSE (p=0.006 to 0.050) and ABX (p=0.045 to 0.050) showed lower gutter areas and volumes compared with VBX. In turn, the VBX showed a nonsignificant tendency to decreased compression, especially compared to ABX. Use of the Excluder C3 showed a 6-fold increase in type A1 gutters (related to type Ia endoleak) as compared to the Conformable Excluder (p=0.018). Balloon-expandable stent-grafts (both ABX and VBX) showed a 3-fold increase in type A1 gutters in comparison with self-expanding stent-grafts (p=0.008). Conclusion: The current study suggests that use of the Conformable Excluder in combination with VSE chimney grafts is superior to the other tested CG/MG combinations in terms of gutter size, gutter type, and CG compression.


Old World cutaneous leishmaniasis treatment response varies depending on parasite species, geographical location and development of secondary infection.
In the Kingdom of Saudi Arabia (KSA), Leishmania major and L. tropica are the main causative agents of Old World cutaneous leishmaniasis (CL). The national CL treatment regimen consists of topical 1% clotrimazole/2% fusidic acid cream followed by 1-2 courses of intralesional sodium stibogluconate (SSG); however, treatment efficacy is highly variable and the reasons for this are not well understood. In this study, we present a complete epidemiological map of CL and determined the efficacy of the standard CL treatment regime in several endemic regions of KSA. Overall, three quarters of patients in all CL-endemic areas studied responded satisfactorily to the current treatment regime, with the remaining requiring only an extra course of SSG. The majority of unresponsive cases were infected with L. tropica. Furthermore, the development of secondary infections (SI) around or within the CL lesion significantly favoured the treatment response of L. major patients but had no effect on L. tropica cases. The response of CL patients to a national treatment protocol appears to depend on several factors, including Leishmania parasite species, geographical location and occurrences of SI. Our findings suggest there is a need to implement alternative CL treatment protocols based on these parameters.


Enhanced estrogen-induced proliferation in obese rat endometrium.
We tested the hypothesis that the proliferative estrogen effect on the endometrium is enhanced in obese vs lean animals. Using Zucker fa/fa obese rats and lean control, we examined endometrial cell proliferation and the expression patterns of certain estrogen-regulated proproliferative and antiproliferative genes after short-term treatment with estradiol. No significant morphologic/histologic difference was seen between the obese rats and the lean rats. Estrogen-induced proproliferative genes cyclin A and c-Myc messenger RNA expression were significantly higher in the endometrium of obese rats compared with those of the lean control. Expression of the antiproliferative gene p27Kip1 was suppressed by estrogen treatment in both obese and lean rats; however, the decrease was more pronounced in obese rats. Estrogen more strongly induced the antiproliferative genes retinaldehyde dehydrogenases 2 and secreted frizzled-related protein 4 in lean rats but had little or no effect in obese rats. Enhancement of estrogen-induced endometrial proproliferative gene expression and suppression of antiproliferative gene expression was seen in the endometrium of obese vs lean animals.


Veno-arterial modified ultrafiltration in children after cardiopulmonary bypass.
A method of performing veno-arterial modified ultrafiltration is described that utilizes conventional blood flow through the aortic and venous cannulae. A dual-pump blood cardioplegia console is adapted to aspirate blood from the cardiopulmonary bypass venous line. The blood is ultrafiltered, sent through the cardioplegia heat exchanger, and returned to the aorta via the cardioplegia needle. Veno-arterial modified ultrafiltration has produced no visual evidence of air entrainment in the cardiopulmonary arterial line. This method allows the immediate resumption of cardiopulmonary bypass without the need for the surgeon to recannulate or alter tubing. Thirty-five children underwent veno-arterial modified ultrafiltration; the results show significant increases in postoperative hematocrit, early extubation, and improved rheology.


Effect of extracellular volume on the energy stored in transmembrane concentration gradients.
The amount of energy that can be retrieved from a concentration gradient across a membrane separating two compartments depends on the relative size of the compartments. Having a larger low-concentration compartment is in general beneficial. It is shown here analytically that the retrieved energy further increases when the high-concentration compartment shrinks during the mixing process, and a general formula is derived for the energy when the ratio of transported solvent to solute varies. These calculations are then applied to the interstitial compartment of the brain, which is rich in Na^{+} and Cl^{-} ions and poor in K^{+}. The reported shrinkage of this compartment, and swelling of the neurons, during oxygen deprivation is shown to enhance the energy recovered from NaCl entering the neurons. The slight loss of energy on the part of K^{+} can be compensated for by the uptake of K^{+} ions by glial cells. In conclusion, the present study proposes that the reported fluctuations in the size of the interstitial compartment of the brain (expansion during sleep and contraction during oxygen deprivation) optimize the amount of energy that neurons can store in, and retrieve from, their ionic concentration gradients.


Past, present and future of the climate and human health commission.
The following paper presents the history of the Climate and Human Health Commission at the International Society of Biometeorology after more than one decade since its creation. A brief history of the origins of the human biometeorology is revealed through some of the main research topics and publications of the founders of the society in this field. Secondly, it is presented as a brief review of the activities of the commission in the last 10 years, based on the reports that have periodically been submitted by members of the commission to the Bulletin of the society. A summary of the topics of interest on human biometeorology and the most frequent research topics are also described. Thirdly, the need of adapting human biometeorology contents, methods and techniques to a changing world is articulated according to some of the new environmental threats in the XXI century. Finally, a list of future actions and research lines collected through a form from members of the commission is presented. The paper concludes with the existence of great challenge for human biometeorology in order to transform biometeorological knowledge into specific services to improve the wellbeing of human beings.


Screening of the venoms of two Parabuthus scorpion species on isolated cardiomyocytes.
Crude, diluted venoms of Parabuthus transvaalicus and P. granulatus were tested on isolated cardiomyocytes. Ventricular myocytes were subjected to hypercontractility tests and whole-cell voltage clamp. Both venoms had a profound influence on the hypercontractility of cardiomyocytes, which was markedly decreased when they were pre-absorbed with commercial antiserum. Whole-cell clamp results showed an increase in the sodium current, and a retardation of the time course of inactivation, implicating the presence of an alpha toxin in both venoms.


Effects of space exposure on ion-beam-deposited silicon-carbide and boron-carbide coatings.
Two recently developed optical coatings, ion-beam-deposited silicon carbide and ion-beam-deposited boron carbide, are very attractive as coatings on optical components for instruments for space astronomy and earth sciences operating in the extreme-UV spectral region because of their high reflectivity, significantly higher than any conventional coating below 105 nm. To take full advantage of these coatings in space applications, it is important to establish their ability to withstand exposure to the residual atomic oxygen and other environmental effects at low-earth-orbit altitudes. The first two flights of the Surface Effects Sample Monitor experiments flown on the ORFEUS-SPAS and the CRISTA-SPAS Shuttle missions provided the opportunity to study the effects of space exposure on these materials. The results indicate a need to protect ion-beam-deposited silicon-carbide-coated optical components from environmental effects in a low-earth orbit. The boron-carbide thin-film coating is a more robust coating able to withstand short-term exposure to atomic oxygen in a low-earth-orbit environment.


Experiences of discrimination or violence and health outcomes among Black, Indigenous and People of Colour trans and/or nonbinary youth.
Identify the relationship between experiences of discrimination or violence and health outcomes for transgender and nonbinary Black, Indigenous and People of Colour (BIPOC) compared to their white trans and nonbinary peers. A national online survey, the 2019 Canadian Trans and Nonbinary Youth Health Survey, was conducted among youth ages 14-25, in English and French. Participants were recruited from November 2018 to May 2019 (N = 1519). BIPOC youth comprised 25.7% of the sample (n = 390). Questions about six types of discrimination (e.g. racism and sexism) and violence (physically threatened or injured), plus foregone health care, self-harm and suicidality were drawn from existing validated measures. Analyses involved cross-tabulations with chi-square tests and logistic regressions. Trans and non-binary BIPOC reported significantly higher prevalence of suicide attempts (24.9% vs. 19.5%) and violence victimization compared to white youth. They had significantly higher odds of self-harm and foregone health care when experiencing discrimination by ethnicity or culture. All types of violence were significantly associated with higher odds of foregone physical health care, self-harm, suicide ideation and suicide attempt. In Canada, trans and nonbinary youth who are BIPOC face disparities in health outcomes and experiences of violence and discrimination compared to white trans and nonbinary youth. Nurses should assess for violence exposure and discrimination among trans and/or nonbinary youth of colour, and promote health equity by advocating for policies to reduce violence and discrimination, including racism, for trans and nonbinary young people.


Sertoli cells require TDP-43 to support spermatogenesis†.
TAR DNA binding protein of 43 kD (TDP-43) is an evolutionarily conserved, ubiquitously expressed transcription factor and RNA-binding protein with major human health relevance. TDP-43 is present in Sertoli and germ cells of the testis and is aberrantly expressed in the sperm of infertile men. Sertoli cells play a key role in spermatogenesis by offering physical and nutritional support to male germ cells. The current study investigated the requirement of TDP-43 in Sertoli cells. Conditional knockout (cKO) of TDP-43 in mouse Sertoli cells caused failure of spermatogenesis and male subfertility. The cKO mice showed decreased testis weight, and low sperm count. Testis showed loss of germ cell layers, presence of vacuoles, and sloughing of round spermatids, suggesting loss of contact with Sertoli cells. Using a biotin tracer, we found that the blood-testis barrier (BTB) was disrupted as early as postnatal day 24 and worsened in adult cKO mice. We noted aberrant expression of the junction proteins connexin-43 (gap junction) and N-cadherin (ectoplasmic specialization). Oil Red O staining showed a decrease in lipid droplets (phagocytic function) in tubule cross-sections, Sertoli cells cytoplasm, and in the lumen of seminiferous tubules of cKO mice. Finally, qRT-PCR showed upregulation of genes involved in the formation and/or maintenance of Sertoli cell junctions as well as in the phagocytic pathway. Sertoli cells require TDP-43 for germ cell attachment, formation and maintenance of BTB, and phagocytic function, thus indicating an essential role for TDP-43 in the maintenance of spermatogenesis.


Two Unusual Cases of Oral Lichen Planus Arising After Oral Squamous Cell Carcinoma: Can Oral Cancer Trigger Autoimmunity?
Autoimmune diseases occur when the immune system fails to recognize self-antigens expressed on the body's own cells and attacks them. Oral lichen planus (OLP) is a chronic autoimmune mucocutaneous disease of the oral cavity characterized by white/red lesions. Considered a potentially malignant disorder, OLP evolution into oral squamous cell carcinoma (OSCC) is still a matter of debate. While chronic autoimmune inflammation is considered a potential risk factor for malignant transformation in many solid tumors, the opposite idea that cancer may trigger autoimmune responses remains controversial. We describe 2 patients who developed lesions clinically suggestive of OLP with histological evidence of lichenoid infiltration some time after OSCC removal, even in areas far from the neoplastic site. Neither patient had OLP before the diagnosis of OSCC, or reported exposure to OLP-associated etiologic factors, and neither. experienced tumor recurrence during follow-up. Our findings suggest that oral cancer remission may be linked to OLP development, but further studies are necessary to unveil the underlying mechanisms and possible prognostic implications.


Externalization of consciousness. Scientific possibilities and clinical implications.
The paper starts by analyzing recent advancements in neurotechnological assessment of residual consciousness in patients with disorders of consciousness and in neurotechnology-mediated communication with them. Ethical issues arising from these developments are described, with particular focus on informed consent. Against this background, we argue for the necessity of further scientific efforts and ethical reflection in neurotechnological assessment of consciousness and 'cerebral communication' with verbally non-communicative patients.


Monensin Enhanced Generation of Extracellular Vesicles as Transfersomes for Promoting Tumor Penetration of Pyropheophorbide-a from Fusogenic Liposome.
The current state of antitumor nanomedicines is severely restricted by poor penetration in solid tumors. It is indicated that extracellular vesicles (EVs) secreted by tumor cells can mediate the intercellular transport of antitumor drug molecules in the tumor microenvironment. However, the inefficient generation of EVs inhibits the application of this approach. Herein, we construct an EV-mediated self-propelled liposome containing monensin as the EV secretion stimulant and photosensitizer pyropheophorbide-a (PPa) as a therapeutic agent. Monensin and PPa are first transferred to the tumor plasma membrane with the help of membrane fusogenic liposomes. By hitchhiking EVs secreted by the outer tumor cells, both drugs are layer-by-layer transferred into the deep region of a solid tumor. Particularly, monensin, serving as a sustainable booster, significantly amplifies the EV-mediated PPa penetration by stimulating EV production. Our results show that this endogenous EV-driven nanoplatform leads to deep tumor penetration and enhanced phototherapeutic efficacy.


Implications of Mobile Genetic Elements for Salmonella enterica Single-Nucleotide Polymorphism Subtyping and Source Tracking Investigations.
Single-nucleotide polymorphisms (SNPs) are widely used for whole-genome sequencing (WGS)-based subtyping of foodborne pathogens in outbreak and source tracking investigations. Mobile genetic elements (MGEs) are commonly present in bacterial genomes and may affect SNP subtyping results if their evolutionary history and dynamics differ from that of the bacterial chromosomes. Using Salmonella enterica as a model organism, we surveyed major categories of MGEs, including plasmids, phages, insertion sequences, integrons, and integrative and conjugative elements (ICEs), in 990 genomes representing 21 major serotypes of S. enterica We evaluated whether plasmids and chromosomal MGEs affect SNP subtyping with 9 outbreak clusters of different serotypes found in the United States in 2018. The median total length of chromosomal MGEs accounted for 2.5% of a typical S. enterica chromosome. Of the 990 analyzed S. enterica isolates, 68.9% contained at least one assembled plasmid sequence. The median total length of assembled plasmids in these isolates was 93,671 bp. Plasmids that carry high densities of SNPs were found to substantially affect both SNP phylogenies and SNP distances among closely related isolates if they were present in the reference genome for SNP subtyping. In comparison, chromosomal MGEs were found to have limited impact on SNP subtyping. We recommend the identification of plasmid sequences in the reference genome and the exclusion of plasmid-borne SNPs from SNP subtyping analysis.IMPORTANCE Despite increasingly routine use of WGS and SNP subtyping in outbreak and source tracking investigations, whether and how MGEs affect SNP subtyping has not been thoroughly investigated. Besides chromosomal MGEs, plasmids are frequently entangled in draft genome assemblies and yet to be assessed for their impact on SNP subtyping. This study provides evidence-based guidance on the treatment of MGEs in SNP analysis for Salmonella to infer phylogenetic relationship and SNP distance between isolates.


Variation amongst survivor populations of white clover collected from sites across Europe: growth attributes and physiological responses to low temperature.
Experiments were carried out at IGER, Aberystwyth, UK to investigate traits of direct relevance to the processes of overwintering and spring growth in white clover (Trifolium repens L.). The plant material used was derived from baseline populations of the cultivar AberHerald and survivor populations generated after 2-3 years' growth in Germany (Kiel), Sweden (Uppsala) and Switzerland (Zürich). The aims of the experiments were to measure the level of genetic shift that had occurred in certain traits due to selection in the survivor populations by comparing these with the baseline population. The adaptive significance of traits was assessed by determining the extent to which stabilizing selection had operated to reduce levels of intra-population variation. Significant differences were found in the responses of leaf production to two temperature treatments in the survivor populations from Germany and Sweden compared with the Swiss and baseline material. Plants of the former two populations produced much more leaf than the others at the higher temperature, but leaf production rates at the lower temperature did not differ. As this experiment used cloned genotypes in the two treatments, the result suggests that a higher degree of phenotypic plasticity for this trait had been selected for in the German and Swedish populations. These populations also showed greater rates of regrowth of leaves from terminal buds exposed to sub-zero temperatures, but there were no differences between populations in levels of freezing tolerance, or in stolon carbohydrate content. Genetic shift occurred in the degree of unsaturation of stolon lipids, with all three survivor populations possessing higher proportions of unsaturated fatty acids than the baseline. Stabilizing selection also operated on this trait in the survivor populations, suggesting that it is of adaptive significance in cool climates.


Allosteric silencing of activating function 1 in the 4-hydroxytamoxifen estrogen receptor complex is induced by substituting glycine for aspartate at amino acid 351.
The active metabolite of tamoxifen, 4-hydroxytamoxifen (4-OHT), is used in the laboratory for mechanistic studies of antiestrogen action. This compound binds to the estrogen receptor alpha (ER) and silences activating function 2 (AF2) in the ligand binding domain, but activating function 1 (AF1) at the other end of the ER remains constitutive and is considered to be ligand independent. Amino acid D351 in the ligand binding domain appears to be critical for interactions with the antiestrogenic side chain of antiestrogens. We have devised an assay to evaluate the biological activity of 351 mutant ERs and antiestrogens at the transforming growth factor alpha (TGFalpha) gene in situ (J. I. MacGregor Schafer et al., Cancer Res., 59: 4308-4313, 1999). The substitution of glycine for aspartate at position 351 results in the conversion of the 4-OHT:ER complex from estrogen-like to completely antiestrogenic. In cells stably expressing D351G ER, the ER retains responsiveness to estradiol (E2) and also retains antiestrogenic responsiveness to both raloxifene and ICI 182,780. The relative binding affinity of E2 for D351G ER (0.77 +/- 0.17 x 10(-9) M) is comparable with wild-type ER (0.42 +/- 0.08 x 10(-9) M). In addition, the D351G ER retains the ability to bind SRC-1 in the presence of E2, thus D351G ER AF2 activity has not been compromised. We also used a cell line stably expressing an ER with a triple mutation in helix 12 (D538A, E542A, and D545A) that ablated AF2 activity, which resulted in decreased effects of E2, suggesting that both AF1 and AF2 activity are required for maximal estrogen activity in MDA-MB-231 cells. Interestingly, the triple mutation also completely reduced the estrogen-like actions of 4-OHT. We propose that a specific mutation at amino acid 351 can allosterically silence AF1 in the 4-OHT:ER complex by either preventing the binding of coactivators or encouraging the binding of a corepressor molecule. We suggest that the 4-OHT-specific site responsible for estrogen-like actions can be referred to as AF2b. This binding site would consist of at least four carboxylic acids at amino acids 351 and 538, 542 and 545 in helix 12 to permit coactivator docking for gene activation. The AF2b site is distinct from AF2 for E2 action. Further studies will provide insight into the estrogen-like actions of tamoxifen in select tissues and breast tumors and identify a significant mechanism of drug resistance to tamoxifen.