Addition of triglyceride to serum for use in quality control and reference.
We describe a simple method in which a water-soluble mixture of triocatanoin and a surfactant, "Triton X-114," are used in preparing solutions of triglycerides (triacylglycerols) in either human serum, solutions of albumin, or water. Analytical recovery added triglyceride was quantitative by two methods. The addition did not affect results of analyses for 18 other commonly measured constitutents of serum. When the triglyceride was added to either lipid-depleted human serum or bovine serum albumin solution and lyophilized, subsequent solutions were clear. The triglyceride/protein preparation was stable in lyophilized form for a year and in reconstituted serum for five days at 5 degrees C. Aquenous solutions appear to be stable indefinitely at room temperature.


Spontaneous hyphaema arising from a strand of persistent pupillary membrane.
A case is reported of spontaneous hyphaema arising from a persistent strand of vascularised pupillary membrane. This strand, and an aneurysmal dilatation from which haemorrhage occurred, were illustrated by fluorescein angiography. The embryogenesis of pupillary membranes is discussed and the literature is reviewed with reference to spontaneous hyphaema.


Flexible Label-Induced Manifold Broad Learning System for Multiclass Recognition.
Broad learning system (BLS), which emerges as a lightweight network paradigm, has recently attracted great attention for recognition problems due to its good balance between efficiency and accuracy. However, the supervision mechanism in BLS and its variants generally relies on the strict binary label matrix, which imposes limitations on approximation and fails to adequately align with the data distribution. To address this issue, in this article, two novel flexible label-induced BLS models with the manifold manner are proposed, whose notable characteristics are as follows. First, two proposed label relaxation strategies can both enlarge the margins between different categories and simultaneously enhance the diversity within labels. Second, the integration of manifold geometrical criterion enables the models to capture local feature structures, ensuring the obtained flexible labels align better with the similarity between samples. Third, the proposed models can be optimized efficiently with the alternating direction method of multipliers. Each iteration benefits from a closed-form solution, facilitating the optimization process. Extensive experiments and thorough theoretical analysis are intended to show the advantages of our proposed models compared to other state-of-the-art recognition algorithms.


Soluble mannosylated myelin peptide inhibits the encephalitogenicity of autoreactive T cells during experimental autoimmune encephalomyelitis.
We have previously shown that immunization with a mannosylated myelin peptide in complete adjuvant induces tolerance instead of disease in experimental autoimmune encephalomyelitis (EAE), a rodent model for multiple sclerosis. In this report we demonstrate that treatment with a soluble mannosylated epitope of proteolipid protein (M-PLP(139-151)) significantly inhibits disease mediated by autoreactive myelin-specific T cells during EAE. Treatment with M-PLP(139-151), applied in different EAE models, significantly reduced the incidence of disease and the severity of clinical symptoms. Delayed-type hypersensitivity responses were abolished after peptide treatment, emphasizing the impact on peripheral T-cell reactivity. Histological analysis of spinal cord tissue from mice treated with M-PLP(139-151) revealed the presence of only few macrophages and T cells. Moreover, little expression of interferon-gamma, interleukin-23, or major histocompatibility complex class II antigen was detected. Immune modulation by M-PLP(139-151) was primarily antigen-specific because an irrelevant mannosylated peptide showed no significant effect on delayed-type hypersensitivity responses or on the course of EAE. Therefore, mannosylated antigens may represent a novel therapeutic approach for antigen-specific modulation of autoreactive T cells in vivo.


Shedding light: a phylotranscriptomic perspective illuminates the origin of photosymbiosis in marine bivalves.
Photosymbiotic associations between metazoan hosts and photosynthetic dinoflagellates are crucial to the trophic and structural integrity of many marine ecosystems, including coral reefs. Although extensive efforts have been devoted to study the short-term ecological interactions between coral hosts and their symbionts, long-term evolutionary dynamics of photosymbiosis in many marine animals are not well understood. Within Bivalvia, the second largest class of mollusks, obligate photosymbiosis is found in two marine lineages: the giant clams (subfamily Tridacninae) and the heart cockles (subfamily Fraginae), both in the family Cardiidae. Morphologically, giant clams show relatively conservative shell forms whereas photosymbiotic fragines exhibit a diverse suite of anatomical adaptations including flattened shells, leafy mantle extensions, and lens-like microstructural structures. To date, the phylogenetic relationships between these two subfamilies remain poorly resolved, and it is unclear whether photosymbiosis in cardiids originated once or twice. In this study, we establish a backbone phylogeny for Cardiidae utilizing RNASeq-based transcriptomic data from Tridacninae, Fraginae and other cardiids. A variety of phylogenomic approaches were used to infer the relationship between the two groups. Our analyses found conflicting gene signals and potential rapid divergence among the lineages. Overall, results support a sister group relationship between Tridacninae and Fraginae, which diverged during the Cretaceous. Although a sister group relationship is recovered, ancestral state reconstruction using maximum likelihood-based methods reveals two independent origins of photosymbiosis, one at the base of Tridacninae and the other within a symbiotic Fraginae clade. The newly revealed common ancestry between Tridacninae and Fraginae brings a possibility that certain genetic, metabolic, and/or anatomical exaptations existed in their last common ancestor, which promoted both lineages to independently establish photosymbiosis, possibly in response to the modern expansion of reef habitats.


In-plane resonant nano-electro-mechanical sensors: a comprehensive study on design, fabrication and characterization challenges.
The newly proposed in-plane resonant nano-electro-mechanical (IP R-NEM) sensor, that includes a doubly clamped suspended beam and two side electrodes, achieved a mass sensitivity of less than zepto g/Hz based on analytical and numerical analyses. The high frequency characterization and numerical/analytical studies of the fabricated sensor show that the high vacuum measurement environment will ease the resonance detection using the capacitance detection technique if only the thermoelsatic damping plays a dominant role for the total quality factor of the sensor. The usage of the intrinsic junction-less field-effect-transistor (JL FET) for the resonance detection of the sensor provides a more practical detection method for this sensor. As the second proposed sensor, the introduction of the monolithically integrated in-plane MOSFET with the suspended beam provides another solution for the ease of resonance frequency detection with similar operation to the junction-less transistor in the IP R-NEM sensor. The challenging fabrication technology for the in-plane resonant suspended gate field-effect-transistor (IP RSG-FET) sensor results in some post processing and simulation steps to fully explore and improve the direct current (DC) characteristics of the sensor for the consequent high frequency measurement. The results of modeling and characterization in this research provide a realistic guideline for these potential ultra-sensitive NEM sensors.


Design of an aryne-platform for the synthesis of non-racemic heterocyclic allocolchicinoids.
A four-step semisynthetic approach towards a highly versatile allocolchicine-related chiral aryne intermediate starting from naturally occurring colchicine was developed, and some of its synthetic transformations were studied. The in situ generated benzyne intermediate afforded a number of non-racemic heterocyclic allocolchicinoids, which were shown to exhibit potent cytotoxicity towards COLO 357, OSA and Raji cells. The proposed methodology is attractive for the synthesis of libraries of new cytotoxic tubulin inhibitors.


The promotion of mental health and the prevention of mental and behavioral disorders: surely the time is right.
This article provides an overview of key issues related to promotion and prevention. It examines the increasing role mental disorders play in the global burden of disease, controversies surrounding constructs such as mental health and prevention, and the links among promotion, prevention, treatment and recovery. After discussing disorders often targeted for prevention, it enumerates principles for effective programs and examples of findings from evidence-based programs. It presents recommendations for the prevention science field and concludes with information about the worldwide response to the need for promotion and prevention services.


Development and Characterization of Nanobody-Derived CD47 Theranostic Pairs in Solid Tumors.
Overexpression of CD47 is frequently observed in various types of human malignancies, inhibiting myeloid-mediated elimination of tumor cells and affecting the prognosis of cancer patients. By mapping biomarker expression, immuno-positron emission tomography has been increasingly used for patient screening and response monitoring. By immunization alpacas with recombinant human CD47, we prepared a CD47-targeting nanobody C2 and developed [68Ga]Ga-NOTA-C2, followed by an exploration of the diagnostic value in CD47-expressing tumor models including gastric-cancer patient-derived xenograft models. By fusing C2 to an albumin binding domain (ABD), we synthesized ABDC2, which had increased in vivo half-life and improved targeting properties. We further labeled ABDC2 with 68Ga/89Zr/177Lu to develop radionuclide theranostic pairs and evaluated the pharmacokinetics and theranostic efficacies of the agents in cell- and patient-derived models. Both C2 and ABDC2 specifically reacted with human CD47 with a high K D value of 23.50 and 84.57 pM, respectively. [68Ga]Ga-NOTA-C2 was developed with high radiochemical purity (99 >%, n = 4) and visualized CD47 expression in the tumors. In comparison to the rapid renal clearance and short half-life of [68Ga]Ga-NOTA-C2, both [68Ga]Ga-NOTA-ABDC2 and [89Zr]Zr-DFO-ABDC2 showed prolonged circulation and increased tumor uptake, with the highest uptake of [89Zr]Zr-DFO-ABDC2 occurring at 72 h post-injection. Moreover, [177Lu]Lu-DOTA-ABDC2 radioimmunotherapy suppressed the tumor growth but was associated with toxicity, warranting further optimization of the treatment schedules. Taken together, we reported a series of nanobody-derived CD47-targeted agents, of which [68Ga]Ga-NOTA-C2 and [89Zr]Zr-DFO-ABDC2 are readily translatable. Optimization and translation of CD47-targeted theranostic pair may provide new prospects for CD47-targeted management of solid tumors.


Evidence indicating proximity in the nucleosome between the histone H4 N termini and the globular domain of histone H1.
Proteolysis of rat liver chromatin by the Arg-C peptidase, clostripain, is characterized by a progressive fragmentation of the N-terminal segments of the four core histones H2A, H2B, H3 and H4, until a well-defined limit digest is reached. This work addresses the case of histone H4. Two intermediate proteolytic sites are identified for this histone, i.e. Arg3 and Arg17, before the limit digest is achieved through cleavage of the polypeptide chain after Arg19. The accessibility of these intermediate sites depends strongly on the presence or absence of histone H1. When H1 is absent, both intermediate sites of histone H4 are similarly accessible, whereas one of them, Arg3, becomes totally inaccessible in the presence of histone H1. Di- and trinucleosomes were used with the aim of avoiding any interference with superstructural effects which can occur with longer polynucleosomes in the presence of H1. We also investigated the accessibility of the Arg sites of H1 that are located primarily in the central globular domain of this histone. In free histone H1, all the centrally located Arg sites are accessible to clostripain. In contrast, in the chromatin-bound state none of these sites is accessible. Besides the arginyl sites in the central globular domain of H1, two Arg residues are observed with the most abundant H1d variant in rat chromatin, one in the N-terminal region and the other in the C-terminal region. The restricted number of proteolytic fragments observed with chromatin-bound H1 is accounted for by the cleavage of H1 after these Arg residues located on the outside of the globular domain. Our results suggest that mutual steric effects are at play between histones H1 and H4 and indicate that the N termini of both histones H4 in the nucleosome lie in close proximity to the globular domain of H1. Based on these observations and taking into account the known structural features of the nucleosome, we propose a model for positioning the N-terminal segments of both histones H4 at the periphery of the nucleoprotein structure. In this model both H4 segments are located within the expanded DNA minor grooves, at periods +/- 1, symmetrically disposed relatively to the nucleosome dyad axis. This arrangement brings the amino ends of both H4 molecules in close contact with the H1 globular domain thus accounting for the observed inaccessibility of the Arg3 site of H4 in the presence of H1.


Potential-Scanning Localized Plasmon Sensing with Single and Coupled Gold Nanorods.
Single plasmonic nanoparticles can potentially serve as optical sensors for detecting local refractive index changes. However, simultaneous monitoring of the scattering spectra from multiple nanoparticles is not practical. Here we perform potential-scanning localized surface plasmon resonance (LSPR) sensing. Gold nanorods are immobilized on an ITO electrode. Instead of collecting the full spectrum, as is done in conventional LSPR sensing, the electrode potential is scanned while the rod spectra are monitored at a single wavelength. We demonstrate that refractive index changes can be determined from single wavelength experiments and we further find that gold nanorod (NR) dimers exhibit higher refractive index sensitivities than single NRs in both potential-scanning and conventional wavelength-scanning based LSPR sensing.


[Identification of nonlinear models of biological membranes by the method of fixed voltage].
A mathematical substantiation of the method suggested by Hodkin and Huxley in 1952 for the identification of nonlinear systems is presented. A procedure for the application of this method was developed, which involves creating the structure of a mathematical model, carrying out a series of tests with specially chosen signals, and finding the unknown parameters. The basic requirements to admissible sets of entrance and target signals and the operator of the system were determined. It was shown that it should be quite continuous, the minimal number of unknown parameters and the minimal complexity of structure of the operator should provide the required quality of approximation. The merits and demerits of the mathematical models of Hodkin-Huxley and Noble, and the procedures used for their creation are discussed. The structure of the operator for the identification of mathematical models of excitable membranes when a large number of membrane currents is considered is offered. It was found that nonlinear electric properties of biological membranes can be identified using tests with other kinds of "fixed" parameters, for example, the method of "fixed" current, the fixed linearly increasing voltage, and others.


Management of idiopathic pulmonary haemosiderosis in pregnancy: report of two cases.
Idiopathic pulmonary haemosiderosis (IPH) is a rare cause of alveolar haemorrhage. Its management in pregnancy is complicated and little clinical data exist on outcomes or treatment. Two patients with IPH, one with a known diagnosis prior to pregnancy (patient A) and one presenting de novo during pregnancy (patient B) are reported. Both subjects became maximally symptomatic in the third trimester and were treated with corticosteroids. Despite steroid treatment patient A had continued symptoms and was treated with azathioprine. Both patients delivered healthy female infants at 32 weeks and 38 weeks gestation, respectively. Patient B's course was complicated by the development of fatal invasive pulmonary aspergillosis 4 weeks postpartum. With improved care, more patients with IPH survive to childbearing age. Patients become most symptomatic late in pregnancy, underscoring the need for careful third trimester monitoring. These cases demonstrate that steroids are safe and that azathioprine can be used as additional therapy. When considering treatment options it is important to weigh the benefits of these medications against the potential for maternal and foetal harm.


[Arterial infusion of combination chemotherapy consisting of adriamycin and mitomycin C for liver metastases of breast cancer].
Forty-nine patients with liver metastases of breast cancer were treated with arterial infusion involving simultaneous use of adriamycin: 30-50 mg and mitomycin C: 10-20 mg through the hepatic artery by Seldinger catheter. Of 45 evaluable patients, there were 5 complete and 12 partial responses (CR + PR: 38%), 21 no change and 7 cases of progressive disease. The median duration of response was 5 months. The median survival time was 7.5 months in all cases, 8.7 months for responders and 6.4 months for non-responders. Leukopenia less than 4 X 10(3)/mm3 was observed in 73% of cases, thrombocytopenia less than 100 X 10(3)/mm3 in 35%, and a decreased hemoglobin value of more than 2 g/dl in 4%, but these were well tolerated. Nausea (53%), vomiting (43%), fever (12%), abdominal pain (6%) and liver toxicity (2%) were observed, but these were manageable. We conclude that this arterial infusion therapy is a useful treatment modality for controlling liver metastases of breast cancer.


Reproduction of inflectional markers in French-speaking children with reading impairment.
Children with reading impairment (RI) experience difficulties in oral and written production of inflectional markers. The origin of these difficulties is not well documented in French. According to some authors, acquisition of irregular items by typically developing children is predicted by token frequency, whereas acquisition of regular items is predicted by type frequency. The authors hypothesized that acquisition of inflectional markers in French depends on the distribution of irregular, invariable, and regular (transparent) items within a grammatical category. Fifteen children with RI age matched with 15 children with typical reading development repeated and read aloud sentences containing adjectives inflected for gender and verbs inflected for number. Inflected adjectives and verbs were matched for token frequency and phonological complexity, whereas distribution of invariable, transparent, and irregular items differed within each grammatical category. Results show higher error rates in the RI group, who produced more errors in reading than repetition, and more errors on inflected verbs than adjectives. Error distribution varied with the proportion of invariable, irregular, and transparent items within each grammatical category, confirming the authors' hypothesis. The authors concluded that morphological difficulties of children with RI group originated from a delay in extracting systematicity in verb and adjective inflectional marking.


Detecting Prolonged Stress in Real Life Using Wearable Biosensors and Ecological Momentary Assessments: Naturalistic Experimental Study.
Increasing efforts toward the prevention of stress-related mental disorders have created a need for unobtrusive real-life monitoring of stress-related symptoms. Wearable devices have emerged as a possible solution to aid in this process, but their use in real-life stress detection has not been systematically investigated. We aimed to determine the utility of ecological momentary assessments (EMA) and physiological arousal measured through wearable devices in detecting ecologically relevant stress states. Using EMA combined with wearable biosensors for ecological physiological assessments (EPA), we investigated the impact of an ecological stressor (ie, a high-stakes examination week) on physiological arousal and affect compared to a control week without examinations in first-year medical and biomedical science students (51/83, 61.4% female). We first used generalized linear mixed-effects models with maximal fitting approaches to investigate the impact of examination periods on subjective stress exposure, mood, and physiological arousal. We then used machine learning models to investigate whether we could use EMA, wearable biosensors, or the combination of both to classify momentary data (ie, beeps) as belonging to examination or control weeks. We tested both individualized models using a leave-one-beep-out approach and group-based models using a leave-one-subject-out approach. During stressful high-stakes examination (versus control) weeks, participants reported increased negative affect and decreased positive affect. Intriguingly, physiological arousal decreased on average during the examination week. Time-resolved analyses revealed peaks in physiological arousal associated with both momentary self-reported stress exposure and self-reported positive affect. Mediation models revealed that the decreased physiological arousal in the examination week was mediated by lower positive affect during the same period. We then used machine learning to show that while individualized EMA outperformed EPA in its ability to classify beeps as originating from examinations or from control weeks (1603/4793, 33.45% and 1648/4565, 36.11% error rates, respectively), a combination of EMA and EPA yields optimal classification (1363/4565, 29.87% error rate). Finally, when comparing individualized models to group-based models, we found that the individualized models significantly outperformed the group-based models across all 3 inputs (EMA, EPA, and the combination). This study underscores the potential of wearable biosensors for stress-related mental health monitoring. However, it emphasizes the necessity of psychological context in interpreting physiological arousal captured by these devices, as arousal can be related to both positive and negative contexts. Moreover, our findings support a personalized approach in which momentary stress is optimally detected when referenced against an individual's own data.


Integrated Dual-Mode Chromatography to Enrich Extracellular Vesicles from Plasma.
Purifying extracellular vesicles (EVs) from complex biological fluids is a critical step in analyzing EVs molecularly. Plasma lipoprotein particles (LPPs) are a significant confounding factor as they outnumber EVs >104 -fold. Given their overlap in size, LPPs cannot be completely removed using standard size-exclusion chromatography. Density-based separation of LPPs can be applied but is impractical for routine use in clinical research and practice. Here a new separation approach, known as dual-mode chromatography (DMC), capable of enriching plasma EVs, and depleting LPPs is reported. DMC conveniently integrates two orthogonal separation steps in a single column device: i) size exclusion to remove high-density lipoproteins (HDLs) that are smaller than EVs; and ii) cation exchange to clear positively charged ApoB100-containing LPPs, mostly (very) low-density lipoproteins (V)LDLs, from negatively charged EVs. The strategy enables DMC to deplete most LPPs (>97% of HDLs and >99% of (V)LDLs) from human plasma, while retaining EVs (>30% of input). Furthermore, the two-in-one operation is fast (15 min per sample) and equipment-free. With abundant LPPs removed, DMC-prepared samples facilitate EV identification in imaging analyses and improve the accuracy for EV protein analysis.


Veno-venous extracorporeal membrane oxygenation (ECMO) for acute respiratory failure caused by liver abscess.
Liver abscess remains a life-threatening disease, particularly when it results in systemic organ failure necessitating intensive care. Only few cases of respiratory failure caused by liver abscess and treated with veno-venous extracorporeal membrane oxygenation (ECMO) have been reported. Here we present a case of liver abscess with rapid progression of multiple organ dysfunction, including severe acute respiratory failure on admission to the intensive care unit (ICU). Upon admission, we immediately initiated artificial organ support systems, including ventilator, continuous renal replacement therapy, and cardiovascular drug infusion for septic multiple organ failure and source control. Despite this initial management, respiratory failure deteriorated and V-V ECMO was introduced. The case developed abdominal compartment syndrome, for which we performed a bedside decompressive laparotomy in the ICU. The case gradually recovered from multiple organ failure and was discharged from the ICU on day 22 and from the hospital on day 53. Since liver abscess is potentially lethal and respiratory failure on admission is an additional risk factor of mortality, V-V ECMO may serve as an adjunctive choice of artificial organ support for cases of severe acute respiratory failure caused by liver abscess.


Ame-miR-980-3p participates in autophagy-mediated midgut remodelling in Apis mellifera via targeting Atg2B.
Autophagy is a process that serves to degrade damaged proteins and organelles, thereby promoting cell homeostasis, differentiation, development and survival. Many miRNAs have been found to have regulatory roles in autophagy. In insects, it has been shown that autophagy is involved in hormone-regulated programmed cell death during metamorphic midgut remodelling. However, whether this is also true during the remodelling of the honey bee midgut is unclear. In the present study, we explored the relationship between autophagy and midgut remodelling and sought to identify miRNAs involved in this physiological process. We found that autophagy occurred during midgut remodelling and that the inhibition of autophagy resulted in midgut dysplasia in prepupae. Differentially expressed miRNAs enriched in the autophagy signalling pathway during midgut remodelling were identified by small RNA-seq. Ame-miR-980-3p, which targets the autophagy-related gene Atg2B, was screened out. Furthermore, abnormal expression of ame-miR-980-3p in the pupal stage led to the thinning of the midgut wall of newly emerged bees (NE). When ame-miR-980-3p expression was inhibited, the intestinal villi of NE bees became significantly shorter and sparse, and the lipid signal in the peritrophic matrix of Pb almost disappeared, indicating that the adult midgut was underdeveloped and the lipid absorption ability was weakened. Taken together, ame-miR-980-3p targeted Atg2B to participate in the regulation of midgut autophagy in the pupae, and the abnormal expression of ame-miR-980-3p would interfere with cell proliferation and death in the process of midgut remodelling, hinder the formation of adult midgut and eventually lead to adult midgut dysplasia and affect the lipid absorption function of the midgut in Apis mellifera.


Up-to-date literature review and issues of sedation during digestive endoscopy.
Sedation is common during digestive endoscopy to provide comfort and pain relief for patients. However, the use of sedation in endoscopy also poses potential risks, and recent issues have been raised regarding its safety and administration. This literature review paper will discuss the most recent developments in the field of sedation in digestive endoscopy, including the adverse events that might be associated with sedation and how to manage it, the legal issues associated with administration, the impact of COVID-19 on sedation practices, and sedation in special situations. It will also touch upon the current guidelines and recommendations for sedation, including the importance of patient selection and monitoring and the need for training and certification for endoscopists administering sedation. The review will also analyse studies evaluating the safety and efficacy of various sedation techniques, including propofol, midazolam, and others. It will examine the benefits and drawbacks of these agents.


Genome sequence and analysis of Lactobacillus helveticus.
The microbiological characterization of lactobacilli is historically well developed, but the genomic analysis is recent. Because of the widespread use of Lactobacillus helveticus in cheese technology, information concerning the heterogeneity in this species is accumulating rapidly. Recently, the genome of five L. helveticus strains was sequenced to completion and compared with other genomically characterized lactobacilli. The genomic analysis of the first sequenced strain, L. helveticus DPC 4571, isolated from cheese and selected for its characteristics of rapid lysis and high proteolytic activity, has revealed a plethora of genes with industrial potential including those responsible for key metabolic functions such as proteolysis, lipolysis, and cell lysis. These genes and their derived enzymes can facilitate the production of cheese and cheese derivatives with potential for use as ingredients in consumer foods. In addition, L. helveticus has the potential to produce peptides with a biological function, such as angiotensin converting enzyme (ACE) inhibitory activity, in fermented dairy products, demonstrating the therapeutic value of this species. A most intriguing feature of the genome of L. helveticus is the remarkable similarity in gene content with many intestinal lactobacilli. Comparative genomics has allowed the identification of key gene sets that facilitate a variety of lifestyles including adaptation to food matrices or the gastrointestinal tract. As genome sequence and functional genomic information continues to explode, key features of the genomes of L. helveticus strains continue to be discovered, answering many questions but also raising many new ones.


Flood insurance in Canada: implications for flood management and residential vulnerability to flood hazards.
Insurance coverage of damage caused by overland flooding is currently not available to Canadian homeowners. As flood disaster losses and water damage claims both trend upward, insurers in Canada are considering offering residential flood coverage in order to properly underwrite the risk and extend their business. If private flood insurance is introduced in Canada, it will have implications for the current regime of public flood management and for residential vulnerability to flood hazards. This paper engages many of the competing issues surrounding the privatization of flood risk by addressing questions about whether flood insurance can be an effective tool in limiting exposure to the hazard and how it would exacerbate already unequal vulnerability. A case study investigates willingness to pay for flood insurance among residents in Metro Vancouver and how attitudes about insurance relate to other factors that determine residential vulnerability to flood hazards. Findings indicate that demand for flood insurance is part of a complex, dialectical set of determinants of vulnerability.


A thiosemicarbazone ligand functionalized by a phosphine group: reactivity toward coinage metal ions.
The reactivity of the phosphino-thiosemicarbazone ligand 2-(2-(diphenylphosphino)benzylidene)-N-ethylthiosemicarbazone (HLPEt) toward M(I) halides (M = Cu, Ag, Au) was studied. The complexes obtained, with formulae [Cu(HLPEt)(2)]Br (1), [Cu(HLPEt)(2)]I.2H(2)O (2), [Ag(HLPEt)(LPEt)].3H(2)O (3), [Au(2)(HLPEt)(2)Cl]Cl.CH(3)OH (4) were satisfactorily characterized by elemental analysis, IR, ESI and (1)H/(31)P NMR. Moreover, we have obtained the crystal structure of the ligand HLPEt and its sulfide oxidized form HLP(S)Et, which represents a new case of desulfurization process in thiosemicarbazone ligands. In addition, we present the crystal structure of the complexes 1 + H(2)O + 3MeOH and [Au(2)(HLPEt)(4)Cl]Cl.3CH(3)OH (5), together with their interesting crystal packing. The complex 5 is an interesting case of a pseudo-chloronium gold(I) complex. The ligand HLPEt and the complexes 3 and 4 display intense luminescence at room temperature.


Spatially dependent lattice deformations for dislocations at the edges of graphene.
We show that dislocations located at the edge of graphene cause different lattice deformations to those located in the bulk lattice. When a dislocation is located near an edge, a decrease in the rippling and increase of the in-plane rotation occurs relative to the dislocations in the bulk. The increased in-plane rotation near the edge causes bond rotations at the edge of graphene to reduce the overall strain in the system. Dislocations were highly stable and remained fixed in their position even when located within a few lattice spacings from the edge of graphene. We study this behavior at the atomic level using aberration-corrected transmission electron microscopy. These results show detailed information about the behavior of dislocations in 2D materials and the strain properties that result.


A randomized EORTC study on the effect of preoperative polychemotherapy in pyriform sinus carcinoma treated by pharyngolaryngectomy and irradiation. Results from 5 to 10 years.
226 patients with an operable epidermoid carcinoma of the pyriform sinus were entered in this clinical trial; 187 cases were considered evaluable, they were distributed as follows: 16 T2, 171 T3, 44 N0, 103 N1 and 40 N3 (UICC TNM Classification 1979). The 89 patients of the chemotherapy group received a 3 day course of Vincristine (1.5 mg/m2), Bleomycin (15 mg), Methotrexate (80 mg). Both the chemotherapy group and the 98 patients of the control group went through the surgical procedure of total laryngectomy with pharyngectomy and radical neck dissection. Radiotherapy was administered postoperatively according to tumor margins and nodal involvement. On examination of the surgical specimen, no evidence of tumor regression was observed in the chemotherapy group. The 3 and 5 year survivals are respectively 45% and 35% in both groups. Node metastasis was studied after N size, N status, capsular rupture (PR +) and revealed early and extensive nodal involvement with 25% capsular ruptures in N0 patients, up to 70% rate in N3. Capsular rupture is also associated with a significantly lower survival and a higher incidence of distant metastases.


5-hydroxymethylfurfural conversion by fungal aryl-alcohol oxidase and unspecific peroxygenase.
Oxidative conversion of 5-hydroxymethylfurfural (HMF) is of biotechnological interest for the production of renewable (lignocellulose-based) platform chemicals, such as 2,5-furandicarboxylic acid (FDCA). To the best of our knowledge, the ability of fungal aryl-alcohol oxidase (AAO) to oxidize HMF is reported here for the first time, resulting in almost complete conversion into 2,5-formylfurancarboxylic acid (FFCA) in a few hours. The reaction starts with alcohol oxidation, yielding 2,5-diformylfuran (DFF), which is rapidly converted into FFCA by carbonyl oxidation, most probably without leaving the enzyme active site. This agrees with the similar catalytic efficiencies of the enzyme with respect to oxidization of HMF and DFF, and its very low activity on 2,5-hydroxymethylfurancarboxylic acid (which was not detected by GC-MS). However, AAO was found to be unable to directly oxidize the carbonyl group in FFCA, and only modest amounts of FDCA are formed from HMF (most probably by chemical oxidation of FFCA by the H2 O2 previously generated by AAO). As aldehyde oxidation by AAO proceeds via the corresponding geminal diols (aldehyde hydrates), the various carbonyl oxidation rates may be related to the low degree of hydration of FFCA compared with DFF. The conversion of HMF was completed by introducing a fungal unspecific heme peroxygenase that uses the H2 O2 generated by AAO to transform FFCA into FDCA, albeit more slowly than the previous AAO reactions. By adding this peroxygenase when FFCA production by AAO has been completed, transformation of HMF into FDCA may be achieved in a reaction cascade in which O2 is the only co-substrate required, and water is the only by-product formed.


Single-cell transcriptomes from human kidneys reveal the cellular identity of renal tumors.
Messenger RNA encodes cellular function and phenotype. In the context of human cancer, it defines the identities of malignant cells and the diversity of tumor tissue. We studied 72,501 single-cell transcriptomes of human renal tumors and normal tissue from fetal, pediatric, and adult kidneys. We matched childhood Wilms tumor with specific fetal cell types, thus providing evidence for the hypothesis that Wilms tumor cells are aberrant fetal cells. In adult renal cell carcinoma, we identified a canonical cancer transcriptome that matched a little-known subtype of proximal convoluted tubular cell. Analyses of the tumor composition defined cancer-associated normal cells and delineated a complex vascular endothelial growth factor (VEGF) signaling circuit. Our findings reveal the precise cellular identities and compositions of human kidney tumors.


Two-Year Outcomes of the New Low-cost Nonvalved Aurolab Aqueous Drainage Implant in Refractory Glaucoma.
This study aimed to report the 2-year surgical outcomes in terms of efficacy and safety of the relatively new, indigenously manufactured, low-cost nonvalved drainage device Aurolab Aqueous Drainage Implant (AADI) in the management of refractory glaucomas in the Indian population. A retrospective review of consecutive patients older than 12 years, who underwent nonvalved glaucoma drainage device surgery via AADI, was performed by a single fellowship-trained surgeon, between January 2014 and January 2018, who had at least 12 months of documented postoperative follow-up. Primary-intraocular pressure (IOP).Secondary-number of antiglaucoma medication (AGM), LogMAR best-corrected visual acuity and complications.Complete success was defined as IOP ≥5 and ≤21 mm Hg; qualified success was defined as fulfilling the above criteria with the use of AGM. Failure was defined as the inability to fulfill the IOP criteria, loss of perception of light, explantation, or any additional glaucoma surgery. A total of 48 eyes of 46 patients were included with a median follow-up of 24 months (quartile 1=16, quartile 3=31.25; interquartile range=15.25) (mean=24.9 mo; SD=10.1). The IOP and number of AGM required was significantly lower at every timepoint postoperatively (P<0.001). The median LogMAR best-corrected visual acuity remained unchanged (P=0.307). Complications occurred in 21 patients (43.7%); some eyes had >1 complication. Complete success was seen in 56.3%; overall success was 87.5%. The safety and efficacy of the new nonvalved glaucoma drainage device, AADI, remain in its second year after implantation. Still further follow-up is recommended, as it has the ability to breach the cost barrier in newly industrialized nations worldwide.


Clinical diagnosis of hyposalivation in hospitalized patients.
The aim of this study was to evaluate the effectiveness of clinical criteria for the diagnosis of hyposalivation in hospitalized patients. A clinical study was carried out on 145 subjects (48 males; 97 females; aged 20 to 90 years). Each subject was clinically examined, in the morning and in the afternoon, along 1 day. A focused anamnesis allowed identifying symptoms of hyposalivation, like xerostomia complaints (considered as a reference symptom), chewing difficulty, dysphagia and increased frequency of liquid intake. Afterwards, dryness of the mucosa of the cheeks and floor of the mouth, as well as salivary secretion during parotid gland stimulation were assessed during oral examination. Results obtained with Chi-square tests showed that 71 patients (48.9%) presented xerostomia complaints, with a significant correlation with all hyposalivation symptoms (p <0.05). Furthermore, xerostomia was also significantly correlated with all data obtained during oral examination in both periods of evaluation (p<0.05). Clinical diagnosis of hyposalivation in hospitalized patients is feasible and can provide an immediate and appropriate therapy avoiding further problems and improving their quality of life.


Perceived added value of a decision support App for formal caregivers in community-based dementia care.
To gain insight into the perceived added value of a decision support App for district nurses and case managers intended to support a problem assessment and the provision of advices on possible solutions to facilitate ageing in place of people with dementia, and to investigate how they would implement the App in daily practice. District nurses and case managers play an important role in facilitating ageing in place of people with dementia (PwD). Detecting practical problems preventing PwD from living at home and advising on possible solutions is complex and challenging tasks for nurses and case managers. To support them with these tasks, a decision support App was developed. A qualitative study using semi-structured interviews was conducted. A photo-elicitation method and an interview guide were used to structure the interviews. The data were analysed according to the principles of content analysis. In five interviews with seven district nurses and case managers, the added value was described in terms of five themes: (a) providing a broader/better overview of possible solutions; (b) providing a guideline/checklist for problem assessment and advice on solutions; (c) supporting an in-depth problem assessment; (d) being a support tool for unexperienced case managers/district nurses; and (e) providing up-to-date information. The participants regarded the App as complementary to their current work procedure, which they would use in a flexible manner at different stages in the care continuum. The participants valued both parts, the problem assessment and the overview of possible solutions. An important requisite for the usage would be that the content is continuously updated. Before implementation of the App can be recommended, an evaluation of its effectiveness regarding decision-making should be conducted. This study underpins the need of nurses and case managers for decision support with regard to problem assessment and providing advices on possible solutions to facilitate ageing in place of PwD. There results also show the importance of listening to users experience and their perceived added value of decision support tools as this helps to explain the lack of statistically significant effects on quantitative outcome measure in contrast to a high willingness to use the App in a previous study.


Small Brain Lesion Enhancement and Gadolinium Deposition in the Rat Brain: Comparison Between Gadopiclenol and Gadobenate Dimeglumine.
The aim of the set of studies was to compare gadopiclenol, a new high relaxivity gadolinium (Gd)-based contrast agent (GBCA) to gadobenate dimeglumine in terms of small brain lesion enhancement and Gd retention, including T1 enhancement in the cerebellum. In a first study, T1 enhancement at 0.1 mmol/kg body weight (bw) of gadopiclenol or gadobenate dimeglumine was evaluated in a small brain lesions rat model at 2.35 T. The 2 GBCAs were injected in an alternated and cross-over manner separated by an interval of 4.4 ± 1.0 hours (minimum, 3.5 hours; maximum, 6.1 hours; n = 6). In a second study, the passage of the GBCAs into cerebrospinal fluid (CSF) was evaluated by measuring the fourth ventricle T1 enhancement in healthy rats at 4.7 T over 23 minutes after a single intravenous (IV) injection of 1.2 mmol/kg bw of gadopiclenol or gadobenate dimeglumine (n = 6/group). In a third study, Gd retention at 1 month was evaluated in healthy rats who had received 20 IV injections of 1 of the 2 GBCAs (0.6 mmol/kg bw) or a similar volume of saline (n = 10/group) over 5 weeks. T1 enhancement of the deep cerebellar nuclei (DCN) was assessed by T1-weighted magnetic resonance imaging at 2.35 T, performed before the injection and thereafter once a week up to 1 month after the last injection. Elemental Gd levels in central nervous system structures, in muscle and in plasma were determined by inductively coupled plasma mass spectrometry (ICP-MS) 1 month after the last injection. The first study in a small brain lesion rat model showed a ≈2-fold higher number of enhanced voxels in lesions with gadopiclenol compared with gadobenate dimeglumine. T1 enhancement of the fourth ventricle was observed in the first minutes after a single IV injection of gadopiclenol or gadobenate dimeglumine (study 2), resulting, in the case of gadopiclenol, in transient enhancement during the injection period of the repeated administrations study (study 3). In terms of Gd retention, T1 enhancement of the DCN was noted in the gadobenate dimeglumine group during the month after the injection period. No such enhancement of the DCN was observed in the gadopiclenol group. Gadolinium concentrations 1 month after the injection period in the gadopiclenol group were slightly increased in plasma and lower by a factor of 2 to 3 in the CNS structures and muscles, compared with gadobenate dimeglumine. In the small brain lesion rat model, gadopiclenol provides significantly higher enhancement of brain lesions compared with gadobentate dimeglumine at the same dose. After repeated IV injections, as expected for a macrocyclic GBCA, Gd retention is minimalized in the case of gadopiclenol compared with gadobenate dimeglumine, resulting in no T1 hypersignal in the DCN.


Dermatokinetics as an Important Tool to Assess the Bioavailability of Drugs by Topical Nanocarriers.
Now-a-days, numerous nanocarrier-based drug products for topical applications are present in the market and number of similar products are being researched. To estimate the amount of drug delivery to skin, the scientists have now established techniques for separation of skin layers for the determination of drug concentrations. This forms the basis of pharmacokinetics of drug(s) in skin, i.e., dermatokinetics. However, dermatokinetic modeling of topical products is still a colossal challenge. Assessment of bioavailability helps in determination of safety and efficacy of topical formulations. This article is an attempt to explore the usefulness and methodologies of dermatokinetics for nanocarriermediated topical delivery. It also showcases challenges in methodologies used for determination of dermatokinetic parameters along with advantages. All the articles (research and review) used for writing the manuscript were collected from various search engines like Science Direct, Google Scholar, PubMed and Eureka Select using keywords like dermatokinetics, novel drug delivery systems, bioequivalence, bioavailability and topical delivery. As the methods used for determination of pharmacokinetics of oral and intravenous formulations are not useful for dermatokinetic assessment, various methods like tape stripping, microdialysis and vasoconstrictor assays are being used for dermatokinetic assessment. These methods are not only useful to determine the drug concentrations in skin layers, but can also be used to correlate the toxic effects of xenobiotics.


Pericardial mesothelioma presenting as chronic constrictive pericarditis. A series of three cases from a single institution.
Pericardial mesotheliomas are rare tumors which often present with features of constrictive pericarditis. We present clinical, imaging, histological, and immunohistochemical findings of three cases presenting with chronic constrictive pericarditis. Two of these cases were initially treated as tuberculous pericarditis. Histologically, all the three cases were of an epithelioid type and showed positivity for more than one mesothelial markers. Two patients had a fatal outcome, and one was lost to follow-up.


Effects of lipoprotein overproduction on the induction of DegP (HtrA) involved in quality control in the Escherichia coli periplasm.
Recent biochemical examination has revealed the presence of at least 90 different lipoproteins in Escherichia coli. Among previously identified lipoproteins, only an outer membrane lipoprotein, NlpE, is known to induce expression of the degP gene upon its overproduction. The degP gene encodes a periplasmic protease, which is thought to be involved in the digestion of unfolded proteins, and is essential for growth at high temperatures. However, it is not completely clear why NlpE overproduction causes degP expression. Moreover, among newly confirmed lipoproteins, there may be others that also induce degP expression. Therefore, we overproduced each of the 90 lipoproteins and examined the level of degP expression as beta-galactosidase activity by using a degP promoter-lacZ fusion. The extent of degP expression caused by NlpE overproduction was dependent on the mode of degP-lacZ fusion. On the other hand, new inner membrane lipoprotein YafY strongly induced degP expression irrespective of the mode of fusion even though the level of overproduced YafY was lower than that of NlpE. The induction of degP expression by YafY overproduction was dependent on the Cpx two-component system. Alteration of the lipoprotein-sorting signals of NlpE and YafY did not abolish the degP induction. However, a YafY derivative possessing the outer membrane signal remained on inner membranes. The non-lipidated derivative of NlpE did not induce degP expression, indicating that membrane anchoring is essential for degP induction. The amino acid sequences of YafY and YfjS, another inner membrane lipoprotein, are highly identical, but overproduction of the latter did not induce degP expression. Construction of various YafY-YfjS chimeric lipoproteins revealed that only a few residues located in the N- and C-terminal regions were important for the induction of DegP.


HIP-I: a huntingtin interacting protein isolated by the yeast two-hybrid system.
We report the discovery of the huntingtin interacting protein I (HIP-I) which binds specifically to the N-terminus of human huntingtin, both in the two-hybrid screen and in in vitro binding experiments. For the interaction in vivo, a protein region downstream of the polyglutamine stretch in huntingtin is essential. The HIP1 cDNA isolated by the two-hybrid screen encodes a 55 kDa fragment of a novel protein. Using an affinity-purified polyclonal antibody raised against recombinant HIP-I, a protein of 116 kDa was detected in brain extracts by Western blot analysis. The predicted amino acid sequence of the HIP-I fragment exhibits significant similarity to cytoskeleton proteins, suggesting that HIP-I and huntingtin play a functional role in the cell filament networks. The HIP1 gene is ubiquitously expressed in different brain regions at low level. HIP-I is enriched in human brain but can also be detected in other human tissues as well as in mouse brain. HIP-I and huntingtin behave almost identically during subcellular fractionation and both proteins are enriched in the membrane containing fractions.


Lesions caused by natural infection with Streptococcus suis type 9 in weaned pigs.
In this paper we report on the spectrum and frequency of lesions observed in a prospective study of naturally occurring infections with Streptococcus suis type 9 (SS-9) in weaned pigs. SS-9 produced arthritis, meningitis, interstitial pneumonia, and endocarditis in 100%, 91%, 73%, and 42%, respectively, of pigs dying from the disease. The lesions were similar to those caused by S. suis type 2 (SS-2), however, SS-9 produced a different frequency of lesions. In SS-2 infections, bronchopneumonia is the most commonly observed lesion; meningitis, arthritis, and endocarditis are less frequently observed. Thus, SS-9 produced a different distribution of lesions from that reported for SS-2. Possible explanations for the differences found are discussed.


GABAA receptor alpha 4 subunits mediate extrasynaptic inhibition in thalamus and dentate gyrus and the action of gaboxadol.
The neurotransmitter GABA mediates the majority of rapid inhibition in the CNS. Inhibition can occur via the conventional mechanism, the transient activation of subsynaptic GABAA receptors (GABAA-Rs), or via continuous activation of high-affinity receptors by low concentrations of ambient GABA, leading to "tonic" inhibition that can control levels of excitability and network activity. The GABAA-R alpha4 subunit is expressed at high levels in the dentate gyrus and thalamus and is suspected to contribute to extrasynaptic GABAA-R-mediated tonic inhibition. Mice were engineered to lack the alpha4 subunit by targeted disruption of the Gabra4 gene. alpha4 Subunit knockout mice are viable, breed normally, and are superficially indistinguishable from WT mice. In electrophysiological recordings, these mice show a lack of tonic inhibition in dentate granule cells and thalamic relay neurons. Behaviorally, knockout mice are insensitive to the ataxic, sedative, and analgesic effects of the novel hypnotic drug, gaboxadol. These data demonstrate that tonic inhibition in dentate granule cells and thalamic relay neurons is mediated by extrasynaptic GABAA-Rs containing the alpha4 subunit and that gaboxadol achieves its effects via the activation of this GABAA-R subtype.


Reagentless substrate analysis with immobilized enzymes.
By coupling an immobilized enzyme system with an electrochemical sensor, the reagent requirement for this glucose method is eliminated. Miniaturization and a further simplification of the instrumentation for the continuous analysis of glucose is achieved.


Detection of oral streptococci in dental unit water lines after therapy with air turbine handpiece: biological fluid retraction more frequent than expected.
Oral streptococci detected in water from dental unit water lines (DUWLs) are a surrogate marker of patients' biological fluid retraction during therapy. We investigated oral streptococci detection rate in DUWLs in a representative sample of private offices in real-life conditions. Samples of nondisinfected water (100 ml) were collected from the DUWL designated for the air turbine handpiece in 81 dental units, immediately after dental treatment of patients with extensive air turbine handpiece use. Water was filtered and plated on a selective medium for oral streptococci and, morphologically, typical colonies of oral streptococci were counted. The lowest detection limit was 0.01 CFU/ml. The oral streptococci detection rate was 72% (95% CI: 62-81%), with a mean level of 0.7 CFU/ml. Oral streptococci detection was not affected by handpiece age or dental treatment type, but was associated with dental unit age. Biological fluid retraction into DUWLs during patient treatment and, possibly, the risk for patient-to-patient blood- or air-borne pathogen transmission are more frequent than expected.


Vitamin B12, folic acid, vitamin D, iron, ferritin, magnesium, and HbA1c levels in patients with diabetes mellitus and dental prosthesis.
Diabetic patients may have vitamin deficiencies, which are important in the follow-up and complications of diabetes for various reasons. It may be beneficial to include the use of dental prosthesis among the parameters that should be investigated as a cause of vitamin deficiency during the management and follow-up of diabetes mellitus. We aimed to investigate the association between serum vitamin B12, folic acid, 25-hydroxyvitamin D, ferritin, iron, magnesium, and HbA1c levels in diabetic patients with and without removable dental prosthesis and in non-diabetic patients with prosthesis. This study is a single-center case-control study. Participants were classified into the following groups: 1) Diabetic patients (n = 528) with prosthesis, 2) non-diabetic patients with prosthesis (n = 121) and 3) diabetic patients without prosthesis (n = 100). Vitamin B12, 25-hydroxyvitamin D, folic acid, ferritin, iron, magnesium, and HbA1c levels were measured and compared across the groups. A significant difference was observed between the groups with respect to the above parameters. Vitamin B12 levels were determined to be higher in the diabetic group without prosthesis. 25-hydroxyvitamin D levels were found to be significantly higher in the non-diabetic group with a prosthesis than in the other two groups. There was no statistical difference in the iron levels between the groups. Ferritin levels were observed to be significantly higher in the diabetic group with prosthesis compared to the other two groups. Magnesium levels were significantly different between all the three groups. The highest magnesium levels were found in the non-diabetic group with prosthesis. HbA1c levels were found to be higher in the diabetic group with prosthesis. Magnesium levels were correlated with 25-hydroxyvitamin D levels, but a negative correlation was observed between these and HbA1c. Serum vitamin B12 levels were lower in the diabetic and non-diabetic groups with prosthesis compared to the diabetic group without prosthesis. 25-hydroxyvitamin D levels were lower and ferritin was higher in the diabetic groups with and without prosthesis. Magnesium levels were significantly lower in the diabetic group with a prosthesis than in the other two groups. The mean HbA1c level was higher in the diabetic group with prosthesis. The comparison of diabetic patients receiving metformin revealed a higher pronounced vitamin B12 deficiency in the diabetic group with prosthesis. These findings show that those diabetic patients with prosthesis should be evaluated for vitamin B12, 25-hydroxyvitamin D, and magnesium deficiency.


Focal therapy versus radical prostatectomy and external beam radiotherapy as primary treatment options for non-metastatic prostate cancer: results of a cost-effectiveness analysis.
Focal therapy treats individual areas of tumour in non-metastatic prostate cancer in patients unsuitable for active surveillance. The aim of this work was to evaluate the cost-effectiveness of focal therapy versus prostatectomy and external beam radiotherapy (EBRT). A Markov cohort health state transition model with four health states (stable disease, local recurrence, metastatic disease and death) was created, evaluating costs and utilities over a 10-year time horizon for patients diagnosed with non-metastatic prostate cancer. National Health Service (NHS) for England perspective was used, based on direct healthcare costs. Clinical transition probabilities were derived from prostate cancer registries in patients undergoing radical prostatectomy, EBRT and focal therapy using cryotherapy (Boston Scientific) or high-intensity focused ultrasound (HIFU) (Sonablate). Propensity score matching was used to ensure that at-risk populations were comparable. Variables included age, prostate-specific antigen (PSA), International Society of Urological Pathology (ISUP) grade group, maximum cancer core length (mm), T-stage and year of treatment. Focal therapy was associated with a lower overall cost and higher quality-adjusted life year (QALY) gains than either prostatectomy or EBRT, dominating both treatment strategies. Positive incremental net monetary benefit (NMB) values confirm focal therapy as cost-effective versus the alternatives at a willingness to pay (WTP) threshold of £30,000/QALY. One-way deterministic sensitivity analyses revealed consistent results. Data used to calculate the transition probabilities were derived from a limited number of hospitals meaning that other potential treatment options were excluded. Limited data were available on later outcomes and none on quality of life data, therefore, literature-based estimates were used. Cost-effectiveness modelling demonstrates use of focal therapy (cryotherapy or HIFU) is associated with greater QALY gains at a lower overall cost than either radical prostatectomy or EBRT, representing good value for money in the NHS.


The development of detour ability during infancy.
3 longitudinal studies were conducted to examine the generalization of detour ability across motor responses and barrier types and the relationship between the development of object permanence and detour ability. In Experiment 1, 12 8-month-olds were tested every 3 weeks for 4 months on 4 different detour problems and Stage 4 and 6 object permanence tasks. In the detour problems, infants had to reach or move around a transparent or opaque barrier to obtain an object. The results indicated that infants made reaching detours before corresponding locomotor ones and generally made detours around opaque barriers before transparent ones. Infants also solved the Stage 4 task before the detour problems but failed to solve the Stage 6 task before testing ended. The results of Experiments 2 and 3 suggested that the difference in reaching and locomotor detour performance was not an artifact of barrier length or the infant's position relative to the barrier. The overall results are discussed in relation to issues of developmental synchrony and Piaget's theory of infant spatial development.


Structure of AMP-PNP-bound BtuCD and mechanism of ATP-powered vitamin B12 transport by BtuCD-F.
The reaction mechanism of BtuCD-F-catalyzed vitamin B12 transport into Escherichia coli is currently unclear. Here we present the structure of the last missing state in the form of AMP-PNP-bound BtuCD, trapped by a disulfide cross-link. Our structural and biochemical data allow a consistent mechanism to be formulated, thus rationalizing the roles of substrate, ATP and substrate-binding protein.


Visceral pain: responses of the reticular formation neurons to gallbladder distension.
Visceral projection (gallbladder distension) to the gigantocellular nucleus of the reticular formation of the cat was tested in neurons classified as pain (P), nonpain-pain (NP-P) and nonpain (NP) units, according to their responses to noxious and/or innocuous natural stimuli from the somatic areas. 96% of P neurons (23 out of 24) responded to gallbladder distension. Quantitative criteria showed comparable effectiveness of the somatic and visceral inputs. NP-P neurons reacted to the gallbladder stimulation in 71% of cases (22 out of 31); NP neurons were activated less effectively. Stimulation of either the central tegmental field or "nonspecific" thalamic nuclei evoked direct responses in 38% of P and 26% of NP-P units, which, in most of the P neurons were followed by excitatory and inhibitory phases. The duration of the latter was approximately one second and it greatly affected the responses of the units to somatic as well as to visceral inputs. A large proportion of P neurons responding to a visceral input documents the important role of the reticular formation in the mechanisms of visceral pain. Findings concerning comparable modifying influences upon reactions of P units both in the case of visceral and painful somatic afferentation indicated that similar control mechanisms could be involved.


Characterization of single cell derived cultures of periosteal progenitor cells to ensure the cell quality for clinical application.
For clinical applications of cells and tissue engineering products it is of importance to characterize the quality of the used cells in detail. Progenitor cells from the periosteum are already routinely applied in the clinics for the regeneration of the maxillary bone. Periosteal cells have, in addition to their potential to differentiate into bone, the ability to develop into cartilage and fat. However, the question arises whether all cells isolated from periosteal biopsies are able to differentiate into all three tissue types, or whether there are subpopulations. For an efficient and approved application in bone or cartilage regeneration the clarification of this question is of interest. Therefore, 83 different clonal cultures of freshly isolated human periosteal cells derived from mastoid periosteum biopsies of 4 donors were generated and growth rates calculated. Differentiation capacities of 51 clonal cultures towards the osteogenic, the chondrogenic, and the adipogenic lineage were investigated. Histological and immunochemical stainings showed that 100% of the clonal cultures differentiated towards the osteogenic lineage, while 94.1% demonstrated chondrogenesis, and 52.9% could be stimulated to adipogenesis. For osteogenesis real-time polymerase chain reaction (PCR) of BGLAP and RUNX2 and for adipogenesis of FABP4 and PPARG confirmed the results. Overall, 49% of the cells exhibited a tripotent potential, 45.1% showed a bipotent potential (without adipogenic differentiation), 3.9% bipotent (without chondrogenic differentiation), and 2% possessed a unipotent osteogenic potential. In FACS analyses, no differences in the marker profile of undifferentiated clonal cultures with bi- and tripotent differentiation capacity were found. Genome-wide microarray analysis revealed 52 differentially expressed genes for clonal subpopulations with or without chondrogenic differentiation capacity, among them DCN, NEDD9, TGFBR3, and TSLP. For clinical applications of periosteal cells in bone regeneration all cells were inducible. For a chondrogenic application a fraction of 6% of the mixed population could not be induced.


[How to measure patient-important outcomes in clinical studies].
Investigators often want to assess patient-important outcomes in clinical studies. They are confronted with the challenging selection of the most appropriate instrument. Instruments should offer an unambiguous and reliable measurement. In addition, there should be evidence from validation studies that they really measure what they intend to measure. Fully standardised questionnaires often fulfil these requirements best and they are amenable to statistical analyses. However it should be ensured that these measurement properties have been verified for the language, in which the questionnaire will be used. There are databases that inform investigators about instruments that have been validated in the respective language.


Effect of cisapride on the post-cholecystectomy upper gastrointestinal transit time.
For the evaluation of postoperative upper gastrointestinal transit, the breath hydrogen test is a convenient method, which is simple but more reliable than the recording of clinical parameters such as bowel sounds. The test was used to study the effect of cisapride on the postoperative mouth-to-caecum transit time. Twenty patients undergoing cholecystectomy were tested pre-operatively and on the first postoperative day after having received either 10 mg of cisapride i.v. or matching placebo under double-blind conditions. The median preoperative transit time in the placebo group was 45 min, and the median postoperative transit time at least 3 h longer. The postoperative transit time after cisapride dosing was still longer than before the operation but was significantly (p = 0.02) reduced as compared with that after placebo administration.


Assessment of psychosocial work hazards and workplace deviant behaviours of teachers in rural community-based schools.
This is a cross-sectional study that assessed the psychosocial work hazards and workplace deviant behaviours of teachers in rural community-based schools. A total of 254 primary school teachers participated in the study. Copenhagen Psychosocial Questionnaire (COPSOQ) and Work Deviance Behaviour Scale were employed to elicit the teachers' responses. The descriptive statistics (frequency and percentage), and Pearson's correlation (r) statistical analysis were used to analyse the data. The results showed that the primary school teachers experience elements of psychosocial work hazards. It was also indicated that primary school teachers rarely exhibit work deviant behaviours. There is a significant positive relationship between psychosocial work hazards and workplace deviant behaviours of primary school teachers in Enugu State. In conclusion, it was suggested that teachers experience psychosocial work hazards and exhibit workplace deviant behaviours in primary schools in Enugu State Nigeria. Therefore, professionals in workplaces including schools should design practicable interventions that would help to reduce workplace deviance among employees in different works of life. With this, these should be reflected in decision-making and policies implementations to rural communities and beyond.


Salinomycin inhibits canine mammary carcinoma in vitro by targeting cancer stem cells.
Salinomycin (SAL), a polyether ionophore antibiotic, has been demonstrated to selectively kill cancer stem cells (CSCs) in various types of human tumor. The aim of the present study was to investigate the effects of SAL on canine mammary CSCs. CSCs in canine mammary carcinoma cell lines (CMT7364 and CIPp) were identified using a sphere formation assay and flow cytometry. The chemoresistance, invasive potential and expression of stem cell-associated proteins of these spheres was then analyzed. This demonstrated that the spheres exhibited characteristics of CSCs, including a cluster of differentiation (CD)44+/CD24-/low phenotype, upregulation of Wnt/β-catenin signaling pathway-associated proteins and chemoresistance. The viability of the spheres was decreased in a concentration- and time-dependent manner following treatment with SAL, and the spheres did not exhibit increased resistance to SAL compared with their parental cells. In addition, exposure to SAL inhibited sphere-formation and invasive potential in canine mammary CSCs in a dose-dependent manner. Furthermore, SAL decreased the CD44+/CD24-/low population and downregulated the expression of Wnt/β-catenin signaling-associated proteins (β-catenin, Cyclin D1 and octamer-binding transcription factor 4) in the spheres. In conclusion, the present study demonstrated that SAL is an effective inhibitor of canine mammary CSCs in vitro, indicating that SAL is a promising chemotherapeutic agent for the treatment of canine mammary carcinoma.


Transcriptional Mechanism of the Mouse β4-Galactosyltransferase 6 Gene in Mouse Neuroblastoma Cell Line Neuro-2a.
Lactosylceramide (Lac-Cer) constitutes the backbone structure of various gangliosides whose abnormal expression is associated with malignancy of neuroblastoma. The understanding of the regulatory mechanism of Lac-Cer contributes to the development of neuroblastoma therapy. In this study, the transcriptional mechanism of mouse β4-galactosyltransferase (β4GalT) 6, which is one of Lac-Cer synthase, was analyzed using mouse neuroblastoma cell line Neuro-2a. The -226 to -13 region relative to the most downstream transcriptional start site was determined to be the promoter region by luciferase assay using the 5'-deletion constructs. The mutation into the activating protein (AP) 4-binding site -110/-101 drastically decreased the promoter activity, indicating that this site is mainly implicated in the transcription. Furthermore, the mutation into the GATA-binding site -210/-201 or another AP4-binding site -202/-193 partially decreased the promoter activity. The study suggests that the mouse β4GalT6 gene is transcriptionally regulated by AP4 in cooperation with GATA family transcription factor in neuroblastoma.


Boletus edulis: a digestion-resistant allergen may be relevant for food allergy.
Fungal components can cause allergic symptoms either through inhalation, ingestion or contact. Whereas respiratory allergy is thought to be induced by spores, allergic reactions following ingestion are attributed to other parts of the mushroom. Reports of food-related allergic reactions due to the edible mushroom Boletus edulis have occasionally been reported. The aim of the study was to investigate whether separate allergens may be detected in alimentary allergy to Boletus edulis. Sera of two subjects, one with recurrent anaphylaxis and the other with a predominantly oral allergy syndrome following ingestion of Boletus edulis, have been analysed by a time-course digestion assay using simulated gastric fluid and by SDS-PAGE immunoblotting. Sera of four Boletus edulis skin prick test-negative subjects and all without clinical symptoms to ingested Boletus edulis served as controls. In lyophilized Boletus edulis extract, at least four water-soluble proteins were detected, the most reactive at 55 kDa and at 80 kDa. Following the time-course digestion assay, IgE binding was found to a 75-kDa protein, but only if the sera of the subject with recurrent anaphylaxis was used. The data indicate that Boletus edulis can cause an IgE-mediated food allergy due to a digestion-stabile protein at 75 kDa. No IgE immune response to this protein was detected in the serum of a subject with respiratory allergy and oral allergy syndrome to Boletus edulis nor in control sera.


Fatty acids. VII. Gas-liquid chromatographic properties of all dimethylene interrupted methyl trans,trans-octadecadienoates.
All the dimethylene interrupted trans,trans-octadecadienoic acids were synthesised by lithium metal-ammonia reduction of the corresponding octadecadiynoic acids, except for the detla2t,6t isomer, which was obtained by the isomerisation of the delta2c,6c isomer. The gas-liquid chromatographic behaviour of the methyl esters was studied on polar [Carbowax 20M, FFAP, DEGA, DEGS, Silar 10C (recently renamed as Apolar 10) and SP 2330], semi-polar (XE-60), and non-polar (SE-30, OV101 and Apiezon L) stationary phases. The equivalent chain length of each isomer is recorded (including the delta2t,6c isomer) and the possibility of separation of these isomers is discussed.


Establishing a System for Effectively Explaining Indications for Driving Prohibition Contained in Drug Package Inserts.
To prevent onset and proliferation of public health/hygiene hazards arising from adverse reactions to drugs, physicians and pharmacists in Japan are now required to provide sufficient explanation of precautions when prescribing or dispensing drugs with package inserts indicating that the patient should not drive, or should take precautions when driving, while taking the drug. Additionally, to ensure proper use of drugs dispensed in accordance with the prescriptions issued by physicians or dentists, the Law for Partial Amendment of the Pharmaceutical Affairs Law (Law No. 103 of 2013) mandates that pharmacists must provide necessary pharmacological research-supported information and guidance to the person receiving the prescribed drug. Under such circumstances, at the Ehime University Hospital Division of Pharmacy we have recently standardized our system for in-person explanation of precautions and necessary guidance to be practiced by pharmacists at our hospital's pharmacy with outpatients being prescribed any drug that requires driving caution. We present concrete examples to support that such standardization of the explanation and guidance system at our hospital has reduced the burden placed on pharmacists in their provision of routine clinical services.


Comparative study on the short-term effects of angiotensin converting enzyme inhibitors (Lopirin, SQUIBB and Tensiomin), and dihydralazine in chronic cardiac failure.
The authors have compared the short-term effect of two captopril (ACE inhibitor) preparations namely the Lopirin (SQUIBB) and Tensiomin (EGIS) and dihydralazine as well as placebo in 15 patients with severe heart failure (NYHA III-IV, class). In case of 8 patients with NYHA IV, functional class the short-term effect of the combined therapy of dihydralazine and Lopirin and dihydralazine and Tensiomin as well dihydralazine and placebo have been compared. The underlying disease was dilated cardiomyopathy (DCM) and ischaemic heart disease (IHD). At the end of the treatment with different drugs and placebo the clinical signs of heart failure (complaints and physical status) and the echo and mechanocardiographic parameters of left ventricular function were assessed. The parameters, apart from the clinical signs, have been evaluated in double blind fashion. Compared to placebo all the three drugs i.e. dihydralazine, Lopirin as well as Tensiomin have decreased significantly the NYHA classes, influenced favorably the non-invasive parameters of left ventricular function and decreased blood pressure. As to the dihydralazine, it improved the left ventricular ejection function and the clinical state of the patients with DCM in a higher degree than the two ACE inhibitors did. The effect of Tensiomin and Lopirin was the same in every respect. Both have influenced more favourable the complaints and physical state of patients with IHD than dihydralazine has. The left ventricular filling pressure, the double product (heart rate x wall tension) indicating the myocardial oxygen demand were more reduced in their effect than in that of dihydralazine. Unlike dihydralazine both decreased the heart rate. Administering one of the two ACE inhibitors to the dihydralazine beneficial additive effects have been experienced; the NYHA classes, the heart rate, the left ventricular wall tension and the double product diminished. The authors, on the bases of the results, consider Tensiomin and Lopirin as equivalent in their effect. In their opinion the administration of these drugs mean a new, efficient way of therapy, first of all in cases of heart failure caused by IHD. In the most severe cases they suggest a trial with the combined dihydralazine-ACE inhibitor therapy.


Immunological Assays used to Support Efficacy of Zika Virus Vaccines.
In February of 2016, the World Health Organization (WHO) declared Zika virus (ZIKV) a Public Health Emergency of International Concern. This prompted a rapid response from both the private and public sector resulting in the generation of several promising vaccine candidates. In this review, we discuss published scientific efforts associated with these novel vaccines, emphasizing the immunological assays used to evaluate their immunogenicity and efficacy, and support future licensure.


Combining genomic analyses with tumour-derived slice cultures for the characterization of an EGFR-activating kinase mutation in a case of glioblastoma.
Epidermal growth factor receptor (EGFR) gene alterations and amplification are frequently reported in cases of glioblastoma (GBM). However, EGFR-activating mutations that confer proven sensitivity to tyrosine kinase inhibitors (TKIs) in lung cancer have not yet been reported in GBM. Using next-generation sequencing, array comparative genomic hybridization and droplet digital PCR, we identified the p.L861Q EGFR mutation in a case of GBM for the first time. The mutation was associated with gene amplification. L861Q may be a clinically valuable mutation because it is known to sensitize non-small-cell lung cancers to treatment with the second-generation EGFR TKI afatinib in particular. Furthermore, we used slice culture of the patient's GBM explant to evaluate the tumour's sensitivity to various EGFR-targeting drugs. Our results suggested that the tumour was not intrinsically sensitive to these drugs. Our results highlight (i) the value of comprehensive genomic analyses for identifying patient-specific, targetable alterations, and (ii) the need to combine genomic analyses with functional assays, such as tumour-derived slice cultures.


Exploring early differentiation and pluripotency in domestic animals.
This short review describes some general features of the origins of the pluripotent inner cell mass and epiblast during the early development of eutherian mammals and the two kinds of embryonic stem cell (ESC), naïve and primed type, that have been produced from these structures. We point out that the derivation of pluripotent stem cells from domesticated species continues to be fraught with difficulties, most likely because the culture requirements of these cells are distinct from those of mouse and human ESCs. Generation of induced pluripotent stem cells (iPSCs) from the domesticated species has been more straightforward, although the majority of the iPSC lines remain dependent on the continued expression of one or more integrated reprogramming genes. Although hope for the potential usefulness of these cells in genetic modification of livestock and other domestic species has dimmed, ESCs and iPSCs remain our best source of self-renewing populations of pluripotent cells, with potential usefulness in preserving and propagating valuable animal breeds and making contributions to fields such as regenerative medicine, toxicology and even laboratory meat production.


Effect of ensiling and silage additives on biogas production and microbial community dynamics during anaerobic digestion of switchgrass.
Silage processing has a crucial positive impact on the methane yield of anaerobic treated substrates. Changes in the characteristics of switchgrass after ensiling with different additives and their effects on methane production and microbial community changes during anaerobic digestion were investigated. After ensiling (CK), methane yield was increased by 33.59% relative to that of fresh switchgrass (FS). In comparison with the CK treatment, methane production was improved by 17.41%, 13.08% and 8.72% in response to ensiling with LBr+X, LBr and X, respectively. A modified Gompertz model predicted that the optimum treatment was LBr+X, with a potential cumulative methane yield of 178.31mL/g total solids (TS) and a maximum biogas production rate of 44.39mL/g TS·d. Firmicutes and Bacteroidetes were the predominant bacteria in FS and silage switchgrass; however, the switchgrass treated with LBr+X was rich in Synergistetes, which was crucial for methane production.


[Polysensory interaction in the cat posterior ventrolateral thalamic nucleus].
The neurone activity was studied in the posterior ventrolateral nucleus of the thalamus in acute experiments on unanesthetized cats. About 20% of the neurons studied proved to react to the visual and somatosensory stimulation. By the character of reaction to these stimuli the polymodal neurons could be divided into three groups. In elaboration of the active defense reflex to visual electric skin stimulation there were recorded in the mentioned nucleus the rhythm assimilation reaction with the action of the flickering light and its depression in case a biologically significant stimulus was presented against this background. After section of the optic tract and separation of the hemispheres to the cerebellar level the rhythm assimilation reaction and its depression were revealed in the posterior ventro-lateral nucleus of the thalamus at the side of the section of the optic tract. This fact indicated that the leading canal of the entrance of visual afferentation to this nucleus passed through the stem reticular formation.


Shared decision making in Italy: An updated revision of the current situation.
The aim of this paper is to update the previous review on the state of patient and public participation in healthcare in Italy. Policymakers consider patient involvement an important aspect in health care decisions and encourage patients to actively participate in the clinical interaction. Nevertheless, the term shared decision making (SDM) is still not clearly defined. Patient associations promote patient participation in health care decisions. Several experts attended the latest consensus conference about patient engagement to reach a consensus on the definition of SDM. Research regarding SDM in Italy continues to increase with 17 articles published between 2012 and 2017. Researchers have assessed the variables associated with patient involvement and explored the use of the SDM approach in different medical settings. Despite the dedicated SDM initiative, researchers in Italy recognize room for improvement. Work is needed to reach a common language regarding SDM and its mechanisms to implement this approach at the clinical level.


Verrucous carcinoma of the bladder with koilocytosis unassociated with vesical schistosomiasis.
Verrucous carcinoma of the bladder is a very rare malignant neoplasm, histologically similar to condyloma acuminatum. Usually it shows association with vesical schistosomiasis (bilharziasis). Only 13 cases unrelated to bilharziasis have been reported to date, and none of them reported koilocytosis, a typical finding in human papillomavirus infection. We report a case of verrucous carcinoma of the bladder that was unrelated to bilharziasis, with koilocytosis and absence of human papillomavirus. The literature relating to the topic is discussed.


Evaluation of subvalvular aortic stenosis in children: a 16-year single-center experience.
Subvalvular aortic stenosis accounts for 1-2 % of all congenital heart disease and for 8-20 % of cases of left-ventricular outflow tract (LVOT) obstruction in children. Recurrence of subaortic stenosis (SAS) is not uncommon after surgical management. This study was performed to investigate the clinical and surgical outcomes and to estimate the predictability of recurrences of SAS. Seventy-nine patients age 3-21 years with SAS between 1994 and 2010 were reviewed. Fifty-one patients had discrete SAS, whereas the remaining 15 patients had fibromuscular ridge-type SAS. Mean follow-up time without surgery was 22 months (range of 1-94). Forty-one patients with a diagnosis of SAS underwent surgery. Recurrence rates were 22.7 % (15 patients), and these patients developed SAS at a mean of 4.7 years follow-up. We performed second surgical membrane resection in only 1 patient. The risk of recurrence of SAS was only linked to higher preoperative LVOT gradient. Twenty-three patients had no aortic regurgitation (AR) at preoperative echocardiography. Of these, 39.1 % had trivial, 8.7 % had mild, and 8.7 % had moderate AR after surgery; there was no significant AR. We conclude that surgical intervention was required most of the time in patients with SAS, and surgical outcomes was excellent even if there were associated cardiac defects. The risk of recurrences was higher, especially in patients with higher initial LVOT gradients, although a second surgery was rarely necessary in these patients.


Developmental expression of three proteins from the first gene of the RNA polymerase sigma 43 operon of Bacillus subtilis.
The first gene of the Bacillus subtilis RNA polymerase sigma 43 operon, P23, has a protein-coding capacity of 23,000 daltons. Sequence analysis revealed three potential translational initiation sites within the same reading frame, which could encode proteins of 23,000 (P23), 19,000 (P19), and 9,000 (P9) daltons, respectively. An internal promoter (P3), which is expressed only during the sporulation stage, is located between the second and the third translational start sites. By protein fusion to the Escherichia coli beta-galactosidase gene, we showed that all three translational initiation sites of the P23 gene are used in vivo in both E. coli and B. subtilis, and regulation for differential expression of the three proteins during the development of B. subtilis is coupled to the transcriptional promoter switching mechanism. The physiological function of these multiple gene products is unknown and is currently under investigation.


Airborne endotoxin associated with particles of different sizes and affected by water content in handled straw.
High exposures to endotoxin are observed in environments where organic materials are handled and lower exposures are found in e.g. indoor air. Inhaled endotoxin contributes significantly to the induction of airway inflammation and dysfunction. The size of an inhaled particle influences the deposition in the airways and the following health symptoms. The objective is to characterise the distribution of endotoxin on airborne particles of different sizes in straw storage halls with high exposure and in other environments with lower exposure levels to endotoxin. Furthermore we have studied the influence of water content of handled straw on the size distribution of endotoxin containing particles. Total, inhalable, thoracic and respirable endotoxin and particles have each been quantified in aerosols from boiler rooms and straw storage halls at 24 power plants, including 21 biofuel plants. Inhalable, thoracic and respirable endotoxin have been quantified in aerosols from offices and outdoor air. The endotoxin concentration was higher in airborne thoracic dust than in airborne 'total dust'. The median respirable fraction in the straw storage halls, boiler rooms at biofuel plants, boiler rooms at conventional plants, offices and outdoors was respectively 42%, 9%, 19%, 24% and 34%. Thoracic endotoxin per number of thoracic particles was higher than respirable endotoxin per number of respirable particles at the biofuel plants. In straw storage halls the fraction of endotoxin of respirable size was highest on the days with lowest water content in the received straw. Furthermore the exposures to all endotoxin fractions were highest on days with the lowest water content in the received straw. In conclusion the highest exposures and concentrations of endotoxin occur or tend to occur from thoracic dust. A high variation in endotoxin concentrations and in fractions of respirable or thoracic size is found in the different working areas. This is important in the risk assessment and makes attempts to influence the endotoxin exposure a possibility. Water content in straw affected the concentration, exposure level and size distribution of airborne endotoxin.


Protectors or Traitors: The Roles of PON2 and PON3 in Atherosclerosis and Cancer.
Cancer and atherosclerosis are major causes of death in western societies. Deregulated cell death is common to both diseases, with significant contribution of inflammatory processes and oxidative stress. These two form a vicious cycle and regulate cell death pathways in either direction. This raises interest in antioxidative systems. The human enzymes paraoxonase-2 (PON2) and PON3 are intracellular enzymes with established antioxidative effects and protective functions against atherosclerosis. Underlying molecular mechanisms, however, remained elusive until recently. Novel findings revealed that both enzymes locate to mitochondrial membranes where they interact with coenzyme Q10 and diminish oxidative stress. As a result, ROS-triggered mitochondrial apoptosis and cell death are reduced. From a cardiovascular standpoint, this is beneficial given that enhanced loss of vascular cells and macrophage death forms the basis for atherosclerotic plaque development. However, the same function has now been shown to raise chemotherapeutic resistance in several cancer cells. Intriguingly, PON2 as well as PON3 are frequently found upregulated in tumor samples. Here we review studies reporting PON2/PON3 deregulations in cancer, summarize most recent findings on their anti-oxidative and antiapoptotic mechanisms, and discuss how this could be used in putative future therapies to target atherosclerosis and cancer.


Occurrence, temporal variation, and estrogenic burden of five parabens in sewage sludge collected across the United States.
Five parabens used as preservatives in pharmaceuticals and personal care products (PPCPs) were measured in sewage sludges collected at 14 U.S. wastewater treatment plants (WWTPs) located in nine states. Detected concentration ranges (ng/g, dry weight) and frequencies were as follows: methyl paraben (15.9 to 203.0; 100%), propyl paraben (0.5 to 7.7; 100%), ethyl paraben (<0.6 to 2.6; 63%), butyl paraben (<0.4 to 4.3; 42%) and benzyl paraben (<0.4 to 3.3; 26%). The estrogenicity inherent to the sum of parabens detected in sewage sludge (ranging from 10.1 to 500.1pg/kg 17β-estradiol equivalents) was insignificant when compared to the 106-times higher value calculated for natural estrogens reported in the literature to occur in sewage sludge. Temporal monitoring at one WWTP provided insights into temporal and seasonal variations in paraben concentrations. This is the first report on the occurrence of five parabens in sewage sludges from across the U.S., and internationally, the first on temporal variations of paraben levels in sewage sludge. Study results will help to inform the risk assessment of sewage sludge destined for land application (biosolids).


Screening the Reference Genes for Quantitative Gene Expression by RT-qPCR During SE Initial Dedifferentiation in Four Gossypium hirsutum Cultivars that Have Different SE Capability.
RNA sequencing (RNA-Seq)-based gene expression analysis is applicable to a wide range of biological purposes in various species. Reverse transcription quantitative PCR (RT-qPCR) is also used to assess target gene expression utilizing stably expressed reference genes as internal control under a given set of conditions. However, investigations of the reference genes for RT-qPCR normalization in the process of somatic embryogenesis (SE) initial dedifferentiation in Gossypium hirsutum are rarely reported. In this study, on the basis of our previous transcriptome data of three different induction stages during SE initial dedifferentiation process in four G. hirsutum cultivars that have different SE capability, 15 candidate genes were selected during SE initial dedifferentiation process, and their expression stability was evaluated by geNorm, NormFinder, and BestKeeper. The results indicated that the two genes of endonuclease 4 (ENDO4) and 18S ribosomal RNA (18S rRNA) showed stable expression in the four different G. hirsutum cultivars, endowing them to be appropriate reference genes during three induction stages in the four cotton cultivars. In addition, the stability and reliability of the two reference genes of ENDO4 and 18S rRNA were further verified by comparing the expressions of auxin-responsive protein 22 (AUX22) and ethylene-responsive transcription factor 17 (ERF17) between RT-qPCR results and the RNA-seq data, which showed strong positive correlation coefficient (R2 = 0.8396-0.9984), validating again the steady expression of ENDO4 and 18S rRNA as the reliable reference genes. Our results provide effective reference genes for RT-qPCR normalization during SE process in different G. hirsutum cultivars.


Abnormalities of immunoregulation in Kawasaki syndrome.
The object of our investigation was to determine the immunoregulatory abnormalities in 48 children with Kawasaki syndrome. We demonstrated a global lymphocytosis with marked expansion of the B cell subset. Despite an increase in B cell numbers, there was a decrease in in vitro immunoglobulin production in response to pokeweed mitogen and hydrocortisone stimulation. These abnormalities correlated with a marked increase in the percentage of CD4+2H4+ (CD4+CD45R+) T cells, a T cell subset thought to be responsible for inducing suppression. Other abnormalities of T cells include cutaneous and in vitro anergy and evidence of T cell activation. Our results suggest that the B cell abnormalities seen in Kawasaki syndrome may be partially explained by defects in T cell immunoregulation.


Potential role of 12 hydroxyeicosatetraenoic acid in angiotensin II-induced calcium signal in rat glomerulosa cells.
Recent evidence suggests that 12 hydroxyeicosatetraenoic acid (12HETE), a product of the 12 lipoxygenase (LO) pathway of arachidonic acid metabolism, may have a role in mediating angiotensin II (AII)-induced aldosterone secretion. The present study examined the possible role of the 12 LO product 12HETE in AII-induced calcium ([Ca++]i) signals in rat glomerulosa cells. The addition of 12HETE to glomerulosa cells induced a dose-dependent (10(-6)-10(-8) M) rise in [Ca++]i levels that was sustained over 15 min. The effects of 12HETE on [Ca++]i were attenuated but not blocked by nifedipine (5 x 10(-6) M) and were preserved in a calcium-free medium, suggesting mobilization of intracellular calcium stores. Furthermore, the 12HETE-mediated rise in [Ca++]i was almost entirely abolished by dantrolene. In parallel, 12HETE reversed the inhibitory effect of nifedipine on AII-induced aldosterone secretion [AII (10(-9) M) - 36 +/- 7, AII + nifedipine (5 x 10(-6) M) - 13 +/- 2, AII + nifedipine + 12HETE (5 x 10(-8) M) - 27 +/- 4 ng/10(6) cells]. Dantrolene also inhibited AII-dependent aldosterone secretion (AII 10(-9) M - 75.8 +/- 5.6, AII + dantrolene 10(-6) M 45.5 +/- 8.8 ng/10(-6) cells), but this inhibition could not be reversed by 12HETE 10(-8) M (45.4 +/- 10.6 ng/10(6) cells). The LO blockers baicalein and BW755C inhibited the effect of AII on aldosterone production and on [Ca++]i in a parallel fashion. During LO blockade, the addition of 12HETE (10(-7) M) restored the AII-induced rise in [Ca++]i. Collectively, these observations suggest that activation of the LO pathway in the rat adrenal glomerulosa contributes to change in cytosolic calcium, which may be important for the steroidogenic effect of AII.


Estimation of material parameters from slow and fast shear waves in an incompressible, transversely isotropic material.
This paper describes a method to estimate mechanical properties of soft, anisotropic materials from measurements of shear waves with specific polarization and propagation directions. This method is applicable to data from magnetic resonance elastography (MRE), which is a method for measuring shear waves in live subjects or in vitro samples. Here, we simulate MRE data using finite element analysis. A nearly incompressible, transversely isotropic (ITI) material model with three parameters (shear modulus, shear anisotropy, and tensile anisotropy) is used, which is appropriate for many fibrous, biological tissues. Both slow and fast shear waves travel concurrently through such a material with speeds that depend on the propagation direction relative to fiber orientation. A three-parameter estimation approach based on directional filtering and isolation of slow and fast shear wave components (directional filter inversion, or DFI) is introduced. Wave speeds of each isolated shear wave component are estimated using local frequency estimation (LFE), and material properties are calculated using weighted least squares. Data from multiple finite element simulations are used to assess the accuracy and reliability of DFI for estimation of anisotropic material parameters.


Iridium-catalyzed enantioselective allylation of silyl enol ethers derived from ketones and α,β-unsaturated ketones.
The unified Ir-catalyzed enantioselective allylic substitution reactions of silyl enol ethers derived from ketones and α,β-unsaturated ketones with branched, racemic allylic alcohols are described. This transformation is catalyzed by the Carreira system and proceeds without fluoride, and with high ee and b : l ratio. The synthetic utility of this method was illustrated by the concise enantioselective total synthesis of marine natural products calyxolane A, B and by the assignment of the absolute configuration of calyxolane A.