Mechanism of membrane fusion induced by vesicular stomatitis virus G protein.
The glycoproteins (G proteins) of vesicular stomatitis virus (VSV) and related rhabdoviruses (e.g., rabies virus) mediate both cell attachment and membrane fusion. The reversibility of their fusogenic conformational transitions differentiates them from many other low-pH-induced viral fusion proteins. We report single-virion fusion experiments, using methods developed in previous publications to probe fusion of influenza and West Nile viruses. We show that a three-stage model fits VSV single-particle fusion kinetics: (i) reversible, pH-dependent, G-protein conformational change from the known prefusion conformation to an extended, monomeric intermediate; (ii) reversible trimerization and clustering of the G-protein fusion loops, leading to an extended intermediate that inserts the fusion loops into the target-cell membrane; and (iii) folding back of a cluster of extended trimers into their postfusion conformations, bringing together the viral and cellular membranes. From simulations of the kinetic data, we conclude that the critical number of G-protein trimers required to overcome membrane resistance is 3 to 5, within a contact zone between the virus and the target membrane of 30 to 50 trimers. This sequence of conformational events is similar to those shown to describe fusion by influenza virus hemagglutinin (a "class I" fusogen) and West Nile virus envelope protein ("class II"). Our study of VSV now extends this description to "class III" viral fusion proteins, showing that reversibility of the low-pH-induced transition and architectural differences in the fusion proteins themselves do not change the basic mechanism by which they catalyze membrane fusion.


Vascular endothelial growth factor inhibition in uveitis: a systematic review.
Vascular endothelial growth factor (VEGF) plays an important role in the pathogenesis of uveitic complications such as cystoid macular oedema (CMO), choroidal neovasularisation (CNV) and retinal neovascularisation (RNV). The use of intravitreal anti-VEGF therapies, namely bevacizumab and ranibizumab, has recently been described in the treatment of these complications. Evidence describing the use of intravitreal anti-VEGF therapy for these complications consists of case reports and case series, most of which are retrospective and have limitations in design and analysis. As such, the current level of evidence supporting the use of intravitreal anti-VEGF therapy for these complications of uveitis would be rated as very low. Furthermore, blockage of VEGF has not been shown to have an anti-inflammatory effect. Thus, treatment of the underlying inflammatory disease should play a central role in the management of uveitic CMO, CNV and RNV. A two-pronged treatment regimen that focuses on achieving disease quiescence through the use of corticosteroids and/or immunosuppressive agents, while treating complications that arise despite adequate disease quiescence with intravitreal anti-VEGF agents, may be useful. However, further data from prospective controlled trials are needed before the therapeutic role of anti-VEGF therapy in the uveitis treatment regimen can be fully determined.


Testin regulates the blood-testis barrier via disturbing occludin/ZO-1 association and actin organization.
The blood-testis barrier (BTB) separates the seminiferous epithelium into the apical and basal compartments. The BTB has to operate timely and accurately to ensure the correct migration of germ cells, meanwhile maintaining the immunological barrier. Testin was first characterized from primary Sertoli cells, it is a secretory protein and a sensitive biomarker to monitor junctions between Sertoli and germ cells. Till now, the functions of testin on BTB dynamics and the involving mechanisms are unknown. Herein, testin acts as a regulatory protein on BTB integrity. In vitro testin knockdown by RNAi caused significant damage to the Sertoli cell barrier with no apparent changes in the protein levels of several major tight junction (TJ), adhesion junction, and gap junction proteins. Also, testin RNAi caused the diffusion of two TJ structural proteins, occludin and ZO-1, diffusing away from the Sertoli cell surface into the cytoplasm. Association and colocalization between ZO-1 and occludin were decreased after testin RNAi, examined by Co-IP and coimmunofluorescent staining, respectively. Furthermore, testin RNAi induced a dramatic disruption on the arrangement of actin filament bundles and a reduced F-actin/G-actin ratio. The actin regulatory protein ARP3 appeared at the Sertoli cell interface after testin RNAi without its protein level change, whereas overexpressing testin in Sertoli cells showed no effect on TJ barrier integrity. The above findings suggest that besides as a monitor for Sertoli-germ cell junction integrity, testin is also an essential molecule to maintain Sertoli-Sertoli junctions.


Effect of health visitors working with elderly patients in general practice: a randomised controlled trial.
Health visitors were employed specifically to care for two years for a random sample of patients in general practice who were aged over 70. Independent assessments made at the beginning and end of the study showed that the health visitor in an urban practice had some impact on her caseload of patients; she provided more services for them, their mortality was reduced, and their quality of life improved, though the last measure just failed to be statistically significant. The health visitor working in a rural practice had no such effect.


Diphosphine Bioconjugates via Pt(0)-Catalyzed Hydrophosphination. A Versatile Chelator Platform for Technetium-99m and Rhenium-188 Radiolabeling of Biomolecules.
The ability to append targeting biomolecules to chelators that efficiently coordinate to the diagnostic imaging radionuclide, 99mTc, and the therapeutic radionuclide, 188Re, can potentially enable receptor-targeted "theranostic" treatment of disease. Here we show that Pt(0)-catalyzed hydrophosphination reactions are well-suited to the derivatization of diphosphines with biomolecular moieties enabling the efficient synthesis of ligands of the type Ph2PCH2CH2P(CH2CH2-Glc)2 (L, where Glc = a glucose moiety) using the readily accessible Ph2PCH2CH2PH2 and acryl derivatives. It is shown that hydrophosphination of an acrylate derivative of a deprotected glucose can be carried out in aqueous media. Furthermore, the resulting glucose-chelator conjugates can be radiolabeled with either 99mTc(V) or 188Re(V) in high radiochemical yields (>95%), to furnish separable mixtures of cis- and trans-[M(O)2L2]+ (M = Tc, Re). Single photon emission computed tomography (SPECT) imaging and ex vivo biodistribution in healthy mice show that each isomer possesses favorable pharmacokinetic properties, with rapid clearance from blood circulation via a renal pathway. Both cis-[99mTc(O)2L2]+ and trans-[99mTc(O)2L2]+ exhibit high stability in serum. This new class of functionalized diphosphine chelators has the potential to provide access to receptor-targeted dual diagnostic/therapeutic pairs of radiopharmaceutical agents, for molecular 99mTc SPECT imaging and 188Re systemic radiotherapy.


Economic and objective burden of caregiving on informal caregivers of patients with systemic vasculitis.
Systemic vasculitis (SV) is associated with substantial economic impact to patients and the healthcare system but little is known about the burden of SV on informal caregivers. We evaluated the objective caregiving burden experienced by informal caregivers of patients with SV. We surveyed adult patients and their informal caregivers on the physical, emotional, social and economic impacts of SV. We asked patients about the extent to which they felt they were a burden to their identified caregivers. Caregivers reported the direct and indirect economic impact of SV, including employment disturbance, income loss and relative time investment of caregiving for their care recipient's SV. We used the Inventory of Caregiving Activities Questionnaire to compute the objective caregiving burden. We analysed data from 68 SV patient-caregiver dyads. Patients reported moderate levels of subjective burden to their caregivers. Over one-quarter of caregivers reported ever having lost some income owing to caregiving for SV. Caregivers reported spending a median of 19 weekly hours on various caregiving tasks, including a median 17 weekly hours on household activities. Given the extended hours that caregivers spend caring for their care recipient, intervention targets should aim to reduce caregiver burnout in the SV population. Future research should examine the relationship between the objective burden of caregiving for SV and the overall physical health, mental health and quality of life of caregivers.


A brief survey on prescriber beliefs regarding compounded topical pain medications.
Pain is extremely common in the U.S. It is estimated that 116 million Americans suffer from chronic pain. Narcotics and opioid medications are heavily relied upon for the treatment of pain. Currently, there is an epidemic of opioid abuse and misuse in the U.S. and alternative methods for the treatment of pain are required. Topical compounded pain creams are thought to be one such alternative. The purpose of this study was to establish a baseline regarding the beliefs of community physicians on the safety and efficacy of these compounds. A survey was sent to practicing physicians in the U.S. regarding their prescribing behaviors for topical compounds, as well as their beliefs about the efficacy and safety of these medications. Eleven prescribing practitioners participated in this survey with a response rate of 78%. The results were compiled with basic statistical work (Excel). This survey demonstrated that at the time of the study, topical compound creams for the treatment of pain were commonly used in clinical practice; these creams were used for musculoskeletal pain and for neuropathic pain. It was also determined that the vast majority of practicing prescribers felt that compounded pain creams were more efficacious than mass-produced, U.S. Food and Drug Administration-approved creams marketed by large multi-national pharmaceutical companies. Most of the respondents felt that these compounds allowed them to reduce their narcotic prescriptions overall. In addition, the results demonstrated that the prescribers felt these compounds were safe for their patients. In this small survey of practicing prescribing providers regarding the use of topical compounds, we found that these medications were not only effective but also safe.


Excited states and two-photon absorption of some novel thiophenyl Pt(II)-ethynyl derivatives.
The photophysical characterization of two new compounds related to bis((4-(phenylethynyl)phenyl)ethynyl)bis(tributylphosphine)platinum(II), here abbreviated Pt1, is reported. For the first new compound ATP1, the inner phenyl rings (closer to the Pt atom) in Pt1 are replaced by thiophene rings bridging at the 2,5-positions. In compound ATP2, the outer phenyl groups are replaced by thiophene rings bonded at the 2-position. Specifically, we report on the fluorescence quantum yield, two-photon absorption, triplet decay times and two-photon absorption induced emission spectra of the molecules in THF solutions. The results were compared with those of Pt1 and Pt1 capped with an acetonide-protected 2,2-bis(methylol)propionic acid (bis-MPA) ester group (Pt1-G1). The photophysical properties of the organic dye 7-(diethylamino)coumarin (Coumarin 110) were determined and used as a reference material. The two-photon absorption cross section around 720-740 nm of ATP1 and ATP2 was found to be of the same order of magnitude as for Pt1-G1, i.e., between 5 and 10 GM, but slightly larger for ATP1 than for ATP2 (1 GM = 1 Göppert-Mayer = 10(-50) (cm(4) s)/photon). The fluorescence decay time of all compounds was found to be very short (subnanosecond) with quantum yields 0.0045, 0.0007, 0.0011 and 0.0020 for ATP1, ATP2, Pt1-G1 and Pt1, respectively, measured at excitation wavelength 373 nm. Just as Pt1 and Pt1-G1, both thiophenyl derivatives showed large intersystem crossing capabilities and phosphorescence, characteristic for a triplet state that can enhance the nonlinear absorption and optical power limiting by triplet state absorption. The phosphorescence emission of the thiophenyl derivatives was red-shifted in comparison with Pt1 and Pt1-G1, and the phosphorescence decay times were on the order of 200-500 ns, as for the Pt1 compound.


The Role of the Central Amygdala in Alcohol Dependence.
Alcohol dependence is a chronically relapsing disorder characterized by compulsive drug-seeking and drug-taking, loss of control in limiting intake, and the emergence of a withdrawal syndrome in the absence of the drug. Accumulating evidence suggests an important role for synaptic transmission in the central nucleus of the amygdala (CeA) in mediating alcohol-related behaviors and neuroadaptive mechanisms associated with alcohol dependence. Acute alcohol facilitates γ-aminobutyric acid (GABA)ergic transmission in the CeA via both pre- and postsynaptic mechanisms, and chronic alcohol increases baseline GABAergic transmission. Acute alcohol inhibits glutamatergic transmission via effects at N-methyl-d-aspartate (NMDA) and α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptors in the CeA, whereas chronic alcohol up-regulates NMDA receptor (NMDAR)-mediated transmission. Pro- (e.g., corticotropin-releasing factor [CRF]) and antistress (e.g., nociceptin/orphanin FQ, oxytocin) neuropeptides affect alcohol- and anxiety-related behaviors, and also alter the alcohol-induced effects on CeA neurotransmission. Alcohol dependence produces plasticity in these neuropeptide systems, reflecting a recruitment of those systems during the transition to alcohol dependence.


The exercise metaboreflex is maintained in the absence of muscle acidosis: insights from muscle microdialysis in humans with McArdle's disease.
1. In McArdle's disease, muscle glycogenolysis is blocked, which results in absent lactate and enhanced ammonia production in working muscle. Using McArdle patients as an experimental model, we studied whether lactate and ammonia could be mediators of the exercise pressor reflex. 2. Changes in muscle interstitial ammonia and lactate were compared with changes in blood pressure and muscle sympathetic nerve activity (MSNA) during static arm flexor exercise at 30% of maximal contraction force. Muscle interstitial changes in lactate and ammonia were assessed by microdialysis of the biceps muscle, and MSNA by peroneal nerve microneurography, in six McArdle patients and 11 healthy, matched controls. One McArdle patient also had myoadenylate deaminase deficiency, a condition associated with abolished ammonia production in exercise. 3. Exercise-induced increases were higher in McArdle patients vs. controls for MSNA (change of 164 +/- 71 vs. 59 +/- 19%) and blood pressure (change of 47 +/- 7 vs. 38 +/- 4 mmHg). Interstitial lactate increased in controls (peak change 1.3 +/- 0.2 mmol x l(-1)) and decreased in McArdle patients (peak change -0.5 +/- 0.1 mmol x l(-1)) during and after exercise. Interstitial ammonia did not change during exercise in either group, but was higher post-exercise in McArdle patients, except in the patient with myoadenylate deaminase deficiency who had a flat ammonia response. This patient had an increase in MSNA and blood pressure comparable to other patients. MSNA and blood pressure responses were maintained during post-exercise ischaemia in both groups, indicating that sympathetic activation was caused, at least partly, by a metaboreflex. 4. In conclusion, changes in muscle interstitial lactate and ammonia concentrations during and after exercise are temporally dissociated from changes in MSNA and blood pressure in both patients with McArdle's disease and healthy control subjects. This suggests that muscle acidification and changes in interstitial ammonia concentration are not mediators of sympathetic activation during exercise.


RLIP, an effector of the Ral GTPases, is a platform for Cdk1 to phosphorylate epsin during the switch off of endocytosis in mitosis.
The Ral signaling pathway is critically involved in Ras-dependent oncogenesis. One of its key actors, RLIP/RalBP1, which participates in receptor endocytosis during interphase, is also involved in mitotic processes when endocytosis is switched off. During mitosis, RLIP76 is located on the duplicated centrosomes and is required for their proper separation and movement to the poles. We have looked for actors that associate with RLIP during mitosis. We show here that RLIP/RalBP1 interacts with an active p34cdc2.cyclinB1 (cdk1) enzyme and that this interaction is crucial for the mitotic phosphorylation of Epsin that, once phosphorylated, is no longer competent for endocytosis. We show also that this latter phosphorylation is dependent on Ral signaling. We propose that RLIP/RalBP1 is used as a platform by the mitotic cdk1 to facilitate the phosphorylation of Epsin, which makes Epsin incompetent for endocytosis during mitosis, when endocytosis is switched off.


Diuretic and natriuretic activity of two mistletoe species in rats.
In different cultural groups, the hemiparasitic plants of the families Loranthaceae and Viscaceae (mistletoes) are frequently used in the treatment of hypertension and/or as diuretic agents. However, it remains unclear as to what commonality makes them diuretic agents or a remedy for hypertension. In this article, the diuretic activity of methanol extracts of Viscum articulatum (VA) Burm. f. and Helicanthus elastica (HE) (Ders.) Dans. in rats is reported. The extracts were administered orally at doses of 100, 200 and 400 mg/kg to rats that had been fasted and deprived of water for 18 hours. Investigations were carried out for diuretic, saluretic and natriuretic effects. The polyphenolic and triterpenoid contents were determined quantitatively using chemical assays and high performance liquid chromatography (HPLC) analysis, respectively. The extracts of VA and HE demonstrated significant and dose-dependent diuretic activity in rats. It was found that while VA mimics the furosemide pattern, HE demonstrated a dose-dependent increase in diuresis, along with an increase in potassium-sparing effects. Phytochemical analysis revealed that polyphenolics and triterpenoids, such as oleanolic acid and lupeol, are the major phytochemicals involved. It was also found that in different combinations, these phytochemicals differed in the way they influenced the electrolyte excretion. A higher content of polyphenolics in association with lower triterpenoid content was found to favor potassium-sparing effects.


Identifying common treatments from Electronic Health Records with missing information. An application to breast cancer.
The aim of this paper is to analyze the sequence of actions in the health system associated with a particular disease. In order to do that, using Electronic Health Records, we define a general methodology that allows us to: (i) identify the actions in the health system associated with a disease; (ii) identify those patients with a complete treatment for the disease; (iii) and discover common treatment pathways followed by the patients with a specific diagnosis. The methodology takes into account the characteristics of the EHRs, such as record heterogeneity and missing information. As an example, we use the proposed methodology to analyze breast cancer disease. For this diagnosis, 5 groups of treatments, which fit in with medical practice guidelines and expert knowledge, were obtained.


Secular trend of earlier onset and decelerated development of third molars: evidence from Croatia.
The purpose of this study was to evaluate the influence of the secular trend on development of third molars in 30 years period in Littoral region of Croatia. A total of 1103 panoramic radiographs of subjects aged from 6 to 18 years were analysed. First group comprised 531 panoramic radiographs (258 girls) taken from year period 1977 till 1979 and second group 572 radiographs (325 girls) taken from year period 2007 till 2009. Demirjian's method was used to determine the developmental stage of third molars. A secular trend in 30 years period was observed as earlier onset of third molars development in boys (6 months on average maxillary and 11 months mandibular) and girls (6 months for maxillary and 4 months for mandibular). Now-a-days, an increase in mineralisation for one Demirjian stage occurs at a slower pace of 5.2 months in boys and 3.4 in girls in maxilla and 2.3 or 2 months in mandible for boys and girls, respectively. The study showed a secular trend present as earlier onset but decelerated development of third molars in both genders and both jaws.


Hot electron field emission via individually transistor-ballasted carbon nanotube arrays.
We present electronically controlled field emission characteristics of arrays of individually ballasted carbon nanotubes synthesized by plasma-enhanced chemical vapor deposition on silicon-on-insulator substrates. By adjusting the source-drain potential we have demonstrated the ability to controllable limit the emission current density by more than 1 order of magnitude. Dynamic control over both the turn-on electric field and field enhancement factor have been noted. A hot electron model is presented. The ballasted nanotubes are populated with hot electrons due to the highly crystalline Si channel and the high local electric field at the nanotube base. This positively shifts the Fermi level and results in a broad energy distribution about this mean, compared to the narrow spread, lower energy thermalized electron population in standard metallic emitters. The proposed vertically aligned carbon nanotube field-emitting electron source offers a viable platform for X-ray emitters and displays applications that require accurate and highly stable control over the emission characteristics.


Biofilm forming abilities of Salmonella are correlated with persistence in fish meal- and feed factories.
Feed contaminated with Salmonella spp. constitutes a risk of Salmonella infections in animals, and subsequently in the consumers of animal products. Salmonella are occasionally isolated from the feed factory environment and some clones of Salmonella persist in the factory environment for several years. One hypothesis is that biofilm formation facilitates persistence by protecting bacteria against environmental stress, e.g. disinfection. The aim of this study was to investigate the biofilm forming potential of Salmonella strains from feed- and fishmeal factories. The study included 111 Salmonella strains isolated from Norwegian feed and fish meal factories in the period 1991-2006 of serovar Agona, serovar Montevideo, serovar Senftenberg and serovar Typhimurium. Significant differences were found between serovars regarding the abilities to form biofilm on polystyrene (microtiter plate assay) and in the air-liquid interface of nutrient broth (pellicle assay). Strains of serovar Agona and serovar Montevideo were good biofilm producers. In Norwegian factories, clones of these serovars have been observed to persist for several years. Most serovar Senftenberg clones appear to persist for a shorter period, and strains of this serovar were medium biofilm producers in our test systems. Strains of the serovar Typhimurium were relatively poor biofilm producers. Salmonella ser. Typhimurium clones have not been observed to persist even though this serovar is resident in Norwegian wild life. When classifying strains according to persistence or presumed non-persistence, persistent strains produced more biofilm than presumed non-persisting strains. The results indicate a correlation between persistence and biofilm formation which suggests that biofilm forming ability may be an important factor for persistence of Salmonella in the factory environment.


Mindfulness approaches to childbirth and parenting.
Mindfulness meditation is increasingly being used as a way of managing pain, reducing stress and anxiety and, in the form of mindfulness-based cognitive therapy (MBCT), as a way of reducing the risk of recurrence in depression (NICE, 2004). This article considers its potential for parents preparing for childbirth focusing on three areas: managing pain during pregnancy and labour; reducing risk of perinatal depression; and increasing 'availability' of attention for the infant. The encouraging evidence to date suggests the possibility that mindfulness has an important contribution to make, both for reducing vulnerability in high-risk groups and as a universal intervention.


Reaction-based Indicator displacement Assay (RIA) for the selective colorimetric and fluorometric detection of peroxynitrite.
Using the self-assembly of aromatic boronic acids with Alizarin Red S (ARS), we developed a new chemosensor for the selective detection of peroxynitrite. Phenylboronic acid (PBA), benzoboroxole (BBA) and 2-(N,N-dimethylaminomethyl)phenylboronic acid (NBA) were employed to bind with ARS to form the complex probes. In particular, the ARS-NBA system with a high binding affinity can preferably react with peroxynitrite over hydrogen peroxide and other ROS/RNS due to the protection of the boron via the solvent-insertion B-N interaction. Our simple system produces a visible colorimetric change and on-off fluorescence response towards peroxynitrite. By coupling a chemical reaction that leads to an indicator displacement, we have developed a new sensing strategy, referred to herein as RIA (Reaction-based Indicator displacement Assay).


Nut and seed consumption and inflammatory markers in the multi-ethnic study of atherosclerosis.
Nuts and seeds are rich in unsaturated fat and other nutrients that may reduce inflammation. Frequent nut consumption is associated with lower risk of cardiovascular disease and type 2 diabetes. The authors examined associations between nut and seed consumption and C-reactive protein, interleukin-6, and fibrinogen in the Multi-Ethnic Study of Atherosclerosis. This 2000 cross-sectional analysis included 6,080 US participants aged 45-84 years with adequate information on diet and biomarkers. Nut and seed consumption was categorized as never/rare, less than once/week, 1-4 times/week, and five or more times/week. After adjustment for age, gender, race/ethnicity, site, education, income, smoking, physical activity, use of fish oil supplements, and other dietary factors, mean biomarker levels in categories of increasing consumption were as follows: C-reactive protein-1.98, 1.97, 1.80, and 1.72 mg/liter; interleukin-6-1.25, 1.24, 1.21, and 1.15 pg/ml; and fibrinogen-343, 338, 338, and 331 mg/dl (all p's for trend < 0.01). Further adjustment for hypertension, diabetes, medication use, and lipid levels yielded similar results. Additional adjustment for body mass index moderately attenuated the magnitude of the associations, yielding borderline statistical significance. Associations of nut and seed consumption with these biomarkers were not modified by body mass index, waist:hip ratio, or race/ethnicity. Frequent nut and seed consumption was associated with lower levels of inflammatory markers, which may partially explain the inverse association of nut consumption with cardiovascular disease and diabetes risk.


Reinforcement learning and human behavior.
The dominant computational approach to model operant learning and its underlying neural activity is model-free reinforcement learning (RL). However, there is accumulating behavioral and neuronal-related evidence that human (and animal) operant learning is far more multifaceted. Theoretical advances in RL, such as hierarchical and model-based RL extend the explanatory power of RL to account for some of these findings. Nevertheless, some other aspects of human behavior remain inexplicable even in the simplest tasks. Here we review developments and remaining challenges in relating RL models to human operant learning. In particular, we emphasize that learning a model of the world is an essential step before or in parallel to learning the policy in RL and discuss alternative models that directly learn a policy without an explicit world model in terms of state-action pairs.


Cell-cell contact and membrane spreading in an ultrasound trap.
An ultrasonic standing wave trap [Langmuir 19 (2003) 3635] in which the morphologies of 2-D latex-microparticle aggregates, forming a pressure node plane, were characterised has been applied here to different cell suspensions with increasing order of specificity of cross-linking molecule, i.e. polylysine with chondrocytes; wheat germ agglutinin (WGA) with erythrocytes and surface receptors on neural cells. The outcome of initial cell-cell contact, i.e. whether the cells stuck at the point of contact (collision efficiency = 1) or rolled around each other (collision efficiency = 0), was monitored in situ by video-microscopy. The perimeter fractal dimensions (FD) of 2-D hexagonally symmetric, closely packed aggregates of control erythrocytes and chondrocytes were 1.16 and 1.18, respectively while those for the dendrititc aggregates formed initially by erythrocytes in 0.5microg/ml WGA and chondrocytes in 20 microg/ml polylysine were 1.49 and 1.66. The FDs for control and molecularly cross-linked cells were typical of reaction-limited aggregation (RLA) and transport diffusion-limited aggregation (DLA), respectively. The FDs of the aggregates of cross-linked cells decreased with time to give more closely packed aggregates without clear hexagonal symmetry. Suspensions of neural cells formed dendritic aggregates. Spreading of inter-cellular membrane contact area occurred over 15 min for both erythrocyte and neural cell dendritic aggregates. The potential of the technique to characterise and control the progression of cell adhesion in suspension away from solid substrata is discussed.


Demographic characteristics of U.S. Chinese older adults in the Greater Chicago area: assessing the representativeness of the PINE study.
We aimed to assess the representativeness of the demographic characteristics of the PINE study to the Chinese aging population in the Greater Chicago area. The PINE study is a population-based study of Chinese older adults aged 60 and over in the Greater Chicago area. In preparation of the PINE study, we conducted a random block census study in Chicago. Demographic characteristics of the PINE study were compared with the data drawn from U.S. Census 2010 and random block census study using chi-square tests. The PINE study is representative of the Chinese aging population in the Greater Chicago area. No significant difference was found in key attributes including age, sex, income, education, number of children, and country of origin. Our report is critical in understanding the vast socio-demographic diversity of U.S. Chinese older adults. Rigorous studies are needed to explore the heterogeneity among the diverse aging populations.


The alternative reading frame tumor suppressor antagonizes hypoxia-induced cancer cell migration via interaction with the COOH-terminal binding protein corepressor.
The alternative reading frame (ARF) tumor suppressor exerts both p53-dependent and p53-independent activities critical to the prevention of cancer in mice and humans. Recent evidence from mouse models suggests that when p53 is absent, further loss of ARF can widen the tumor spectrum, and potentiate invasion and metastasis. A major target of the p53-independent activity of ARF is the COOH-terminal binding protein (CtBP) family of metabolically regulated transcriptional corepressors, which are degraded upon acute exposure to the ARF protein. CtBPs are activated under conditions of metabolic stress, such as hypoxia, to repress epithelial and proapoptotic genes, and can mediate hypoxia-induced migration of cancer cells. The possibility that ARF could suppress tumor cell migration as part of its p53-independent activities was thus explored. Small-interfering RNA (siRNA)-mediated knockdown of ARF in human lung carcinoma cells led to increased cell migration, especially during hypoxia, and this effect was blocked by concomitant treatment with CtBP2 siRNA. Introduction of ARF into p53 and ARF-null human colon cancer cells inhibited hypoxia-induced migration. Furthermore, overexpression of CtBP2 in ARF-expressing cells enhanced cell migration, and an ARF mutant defective in CtBP-family binding was impaired in its ability to inhibit cell migration induced by CtBP2. ARF depletion or CtBP2 overexpression was associated with decreased PTEN expression and activation of the phosphatidylinositol 3-kinase pathway, and a phosphatidylinositol 3-kinase inhibitor blocked CtBP2-mediated cell migration. Thus, ARF can suppress cell migration by antagonizing CtBP2 and the phosphatidylinositol 3-kinase pathway, and these data may explain the increased aggressiveness of ARF-null tumors in mouse models.


Evaluation of the organised mammographic screening programme in Australia.
An organised approach to mammographic screening in Australia commenced in 1992 as a free service for asymptomatic women in the target age range 50-69 years and accessible to all women aged >/=40 years. Screening is performed by two-view mammography, women aged 50-69 years being sent reminders every 2 years for repeat screens. National Accreditation Standards have been agreed and BreastScreen Australia has developed mechanisms to monitor and report performance including systems and standards for data collection. The national report includes participation rates, small cancer detection rates and programme sensitivities. These indicate that BreastScreen Australia is meeting its accreditation standards with respect to most measures. Routinely collected outcome data reflect an increasing trend in the proportion of women receiving breast conserving surgery; a decline in the proportion of women diagnosed with ductal carcinoma in situ undergoing axillary dissection and an increasing use of combination therapies with a corresponding decrease in the use of hormonal therapy alone. There have been increases in breast cancer incidence and falls in mortality with the largest changes in women aged 50-69 years. As the BreastScreen Australia database is routinely linked in future to population cancer registries, the mortality reduction due to mammographic screening may be estimated more precisely.


Emotional intelligence: a primal dimension of nursing leadership?
Emotional intelligence, a concept well known in the world of business, is quickly gain ing recognition in the healthcare arena. Described as the primal aspect of leadership, emotional intelligence refers to the ability to monitor and discriminate among emotions, and to use the information to guide thought and action. This paper will explore the theoretical and empirical basis of emotional intelligence and its linkages to leadership. Readers will be informed of the origins and theoretical perspectives regarding emotional intelligence, evidence related to the existence of emotional intelligence, benefits and limitations of the concept and implications for nursing leadership.


Hypertension as related to renal function in diabetes mellitus.
The relationships between blood pressure and renal function were investigated in 78 hypertensive patients with diabetes mellitus type I or II. Renal function was assessed by determining the glomerular filtration rate and the para-aminohippurate clearance in 32 and serum creatinine in 46 subjects. In the latter, the reciprocal serum creatinine, corrected for age and changing creatinine/insulin clearance ratio, was used as an estimate of glomerular filtration rate. In the 54 patients with serial determinations, the duration of follow-up averaged 10.5 years. In older patients with type II diabetes without clinical proteinuria, hypertension developed either before or after the onset of diabetes. When it appeared, renal function was only slightly reduced. During follow-up, the decline in reciprocal serum creatinine averaged 2.7% per year, a figure very similar to that found in nondiabetic patients with benign essential hypertension. It did not correlate with the blood pressure. In patients with a proteinuria greater than 2.5 g per day and histologic and/or clinical evidence of diabetic glomerulosclerosis, the severity of hypertension correlated inversely with the level of renal function. The rate of decline in function averaged 11% per year but varied widely. It was not significantly related to the blood pressure. These data suggest that different types of hypertension (essential, diabetic, and nephrogenic) may be associated with diabetes mellitus. The rate of decline in renal function is closely related to the presence or absence of clinical proteinuria but not to the level of blood pressure.


Transcranial Doppler ultrasonography to confirm brain death: a meta-analysis.
Barbiturate therapy or hypothermia precludes proper diagnosis of brain death either clinically or by EEG. Specific intracranial flow patterns indicating cerebral circulatory arrest (CCA) can be visualized by transcranial Doppler ultrasonography (TCD). The aim of this study was to assess the validity of TCD in confirming brain death. Meta-analysis of studies assessing the validity of TCD in confirming brain death. A systematic review of articles in English on the diagnosis brain death by TCD, published between 1980 and 2004, was performed. An oscillating or reverberating flow and systolic spikes were considered to be compatible with CCA. The quality of each study was assessed using standardized methodological criteria. The literature was searched for any article reporting a false-positive result. Two high-quality and eight low-quality studies were included. Meta-analysis of the two high-quality studies showed a sensitivity of 95% (95% CI 92-97%) and a specificity of 99% (95% CI 97-100%) to detect brain death. Meta-analysis of all ten studies showed a sensitivity of 89% and a specificity of 99%. In the literature we found two false-positive results; however, in both patients brain-stem function did show brain death shortly thereafter. CCA by TCD in the anterior and posterior circulation predicted fatal brain damage in all patients; therefore, TCD can be used to determine the appropriate moment for angiography. Further research is needed to demonstrate that CCA by TCD on repeated examination can also predict brain death in all patients.


Functional anatomy of the masking level difference, an fMRI study.
Masking level differences (MLDs) are differences in the hearing threshold for the detection of a signal presented in a noise background, where either the phase of the signal or noise is reversed between ears. We use N0/Nπ to denote noise presented in-phase/out-of-phase between ears and S0/Sπ to denote a 500 Hz sine wave signal as in/out-of-phase. Signal detection level for the noise/signal combinations N0Sπ and NπS0 is typically 10-20 dB better than for N0S0. All combinations have the same spectrum, level, and duration of both the signal and the noise. Ten participants (5 female), age: 22-43, with N0Sπ-N0S0 MLDs greater than 10 dB, were imaged using a sparse BOLD fMRI sequence, with a 9 second gap (1 second quiet preceding stimuli). Band-pass (400-600 Hz) noise and an enveloped signal (.25 second tone burst, 50% duty-cycle) were used to create the stimuli. Brain maps of statistically significant regions were formed from a second-level analysis using SPM5. The contrast NπS0- N0Sπ had significant regions of activation in the right pulvinar, corpus callosum, and insula bilaterally. The left inferior frontal gyrus had significant activation for contrasts N0Sπ-N0S0 and NπS0-N0S0. The contrast N0S0-N0Sπ revealed a region in the right insula, and the contrast N0S0-NπS0 had a region of significance in the left insula. Our results extend the view that the thalamus acts as a gating mechanism to enable dichotic listening, and suggest that MLD processing is accomplished through thalamic communication with the insula, which communicate across the corpus callosum to either enhance or diminish the binaural signal (depending on the MLD condition). The audibility improvement of the signal with both MLD conditions is likely reflected by activation in the left inferior frontal gyrus, a late stage in the what/where model of auditory processing.


The Americleft Psychosocial Outcomes Project: A Multicenter Approach to Advancing Psychosocial Outcomes for Youth With Cleft Lip and Palate.
Cleft lip and/or palate (CL/P) are among the most common of all birth defects. Habilitation requires multiple surgeries and other therapies throughout childhood and adolescence. While multidisciplinary care is recommended, there is a great deal of variation in treatment protocols for this condition. There is ample evidence that children with CL/P are at risk for psychosocial problems. However, to date, few studies have systematically investigated parent and patient self-reported psychosocial and quality of life (QOL) outcomes for children with CL/P as they relate to variations in treatment protocols. The Americleft Outcomes project was initiated to demonstrate and document outcomes to be expected with team care, and to define the key features or characteristics of various team treatment protocols and procedures that are associated with more or less favorable/desirable outcomes. This article will describe the psychosocial component of the Americleft Outcomes project that is aimed at developing a protocol that will allow cross team assessment of psychosocial outcomes for children with CL/P in relationship to the treatments they received. The protocol will be detailed along with a description of the process and considerations that were instrumental in the development of the project. Stakeholder input about the project's perceived relevance to families of children with CL/P will be reported. The paper concludes with a discussion of the challenges encountered with this project, clinical implications, and future directions.


Reaction screening and optimization of continuous-flow atropine synthesis by preparative electrospray mass spectrometry.
Preparative electrospray (ES) exploits the acceleration of reactions in charged microdroplets to perform a small scale chemical synthesis. In combination with on-line mass spectrometric (MS) analysis, it constitutes a rapid screening tool to select reagents to generate specific products. A successful reaction in preparative ES triggers a refined microfluidic reaction screening procedure which includes the optimization for stoichiometry, temperature and residence time. We apply this combined approach for refining a flow synthesis of atropine. A successful preparative ES pathway for the synthesis of the phenylacetyl ester intermediate, using tropine/HCl/phenylacetyl chloride, was optimized for solvent in both the preparative ES and microfluidics flow systems and a base screening was conducted by both methods to increase atropine yield, increase percentage conversion and reduce byproducts. In preparative ES, the first step yielded 55% conversion (judged using MS) to intermediate and the second step yielded 47% conversion to atropine. When combined in two discrete steps in continuous-flow microfluidics, a 44% conversion of the starting material and a 30% actual yield of atropine were achieved. When the reactions were continuously telescoped in a new form of preparative reactive extractive electrospray (EES), atropine was synthesized with a 24% conversion. The corresponding continuous-flow microfluidics experiment gave a 55% conversion with an average of 34% yield in 8 min residence time. This is the first in depth study to utilize telescoped preparative ES and the first use of dual ESI emitters for multistep synthesis.


"Inverse Drug Discovery" Strategy To Identify Proteins That Are Targeted by Latent Electrophiles As Exemplified by Aryl Fluorosulfates.
Drug candidates are generally discovered using biochemical screens employing an isolated target protein or by utilizing cell-based phenotypic assays. Both noncovalent and covalent hits emerge from such endeavors. Herein, we exemplify an "Inverse Drug Discovery" strategy in which organic compounds of intermediate complexity harboring weak, but activatable, electrophiles are matched with the protein(s) they react with in cells or cell lysate. An alkyne substructure in each candidate small molecule enables affinity chromatography-mass spectrometry, which produces a list of proteins that each distinct compound reacts with. A notable feature of this approach is that it is agnostic with respect to the cellular proteins targeted. To illustrate this strategy, we employed aryl fluorosulfates, an underexplored class of sulfur(VI) halides, that are generally unreactive unless activated by protein binding. Reversible aryl fluorosulfate binding, correct juxtaposition of protein side chain functional groups, and transition-state stabilization of the S(VI) exchange reaction all seem to be critical for conjugate formation. The aryl fluorosulfates studied thus far exhibit chemoselective reactivity toward Lys and, particularly, Tyr side chains, and can be used to target nonenzymes (e.g., a hormone carrier or a small-molecule carrier protein) as well as enzymes. The "Inverse Drug Discovery" strategy should be particularly attractive as a means to explore latent electrophiles not typically used in medicinal chemistry efforts, until one reacts with a protein target of exceptional interest. Structure-activity data can then be used to enhance the selectivity of conjugate formation or the covalent probe can be used as a competitor to develop noncovalent drug candidates. Here we use the "Inverse Drug Discovery" platform to identify and validate covalent ligands for 11 different human proteins. In the case of one of these proteins, we have identified and validated a small-molecule probe for the first time.


Symmetry-controlled colloidal nanocrystals: nonhydrolytic chemical synthesis and shape determining parameters.
Since inorganic nanocrystals exhibit unique shape-dependent nanoscale properties and can be utilized as basic building blocks for futuristic nanodevices, a systematic study on the shape control of these nanocrystals remains an important subject in materials and physical chemistry. In this feature article, we overview the recent progress on the synthetic development of symmetry-controlled colloidal nanocrystals of semiconductor and metal oxide, which are prepared through nonhydrolytic chemical routes. We describe their shape-guiding processes and illustrate the detailed key factors controlling their growth by examining various case studies of zero-dimensional spheres and cubes, one-dimensional rods, and quasi multidimensional structures such as disks, multipods, and stars. Specifically, the crystalline phase of nucleating seeds, surface energy, kinetic vs thermodynamic growth, and selective adhesion processes of capping ligands are found to be most crucial for the determination of the nanocrystal shape.


The changes of the number and regional distribution of dentists and dental institutions 9 years after the implementation of postgraduate year training program for dentists in Taiwan.
In Taiwan, there are three major dentist occupation problems: a surplus of dentists, an uneven regional distribution of dentists, and a concentration of dentists in the metropolitan areas. The purpose of this study was to evaluate the distributions of the population and dentists in each city or county in 2001, 2010 and 2019 to further analyze the changes of regional distributions of dentists 9 years after the implementation of postgraduate year training program for dentists (PGYD) in 2010 in Taiwan. This study collected the data of population, the number of dentists in each city or county in 2001, 2010 and 2019, and the numbers of dentists and dental institutions from 2001 to 2019 for evaluating the changes of regional distributions of dentists 9 years after the PGYD implementation in Taiwan. Nine years after the PGYD implementation, the uneven distribution of dentists persisted. The number of dentists still increased and dentists were more concentrated in municipalities than in non-municipalities in Taiwan. Moreover, the number of dental institutions increased slowly, but dental institutions tended to become larger. Because the PGYD system is not mandatory, dentists choose their practice locations as driven by the market factors. The increase in the average number of dentists in dental institutions is also driven by the market factors. The market-driven movement of dentists to the people-concentrated municipalities finally persist the imbalance of dentist distribution in Taiwan. This situation also enables the development of large-scale dental institutions in municipalities in Taiwan.


Patterns and Predictors of Relapse Following Radical Chemoradiation Therapy Delivered Using Intensity Modulated Radiation Therapy With a Simultaneous Integrated Boost in Anal Squamous Cell Carcinoma.
Our purpose was to describe the patterns and predictors of treatment failure in patients receiving definitive chemoradiation therapy (CRT) for anal squamous cell carcinoma (ASCC), delivered using intensity modulated radiation therapy (IMRT). Our study was a retrospective cohort analysis of consecutive patients treated with curative intent for ASCC using CRT delivered with a standardized IMRT technique in 5 UK cancer centers. Patients were included from the start of UK IMRT guidance from February 2013 to October 31, 2017. Collected data included baseline demographics, treatment details, tumor control, sites of relapse, and overall survival. Statistical analysis to calculate outcomes and predictive factors for outcome measures were performed using SPSS and R. The medical records of 385 consecutive patients were analyzed. Median follow-up was 24.0 months. Within 6 months of completing CRT, 86.7% of patients achieved a complete response. Three-year disease-free survival and overall survival were 75.6% and 85.6%, respectively. Of all relapses, 83.4% occurred at the site of primary disease. There were 2 isolated relapses in regional nodes not involved at outset. Predictive factors for cancer recurrence included male sex, high N-stage, and failure to complete radiation therapy as planned. The treatment results compare favorably to published outcomes from similar cohorts using 3-dimensional conformal CRT. The observed patterns of failure support the current UK IMRT voluming guidelines and dose levels, highlighting our prophylactic nodal dose as sufficient to prevent isolated regional relapse in uninvolved nodes. Further investigation of strategies to optimize CR should remain a priority in ASCC because the site of primary disease remains the overwhelming site of relapse.


Combinatorial Drug Screening Identifies Ewing Sarcoma-specific Sensitivities.
Improvements in survival for Ewing sarcoma pediatric and adolescent patients have been modest over the past 20 years. Combinations of anticancer agents endure as an option to overcome resistance to single treatments caused by compensatory pathways. Moreover, combinations are thought to lessen any associated adverse side effects through reduced dosing, which is particularly important in childhood tumors. Using a parallel phenotypic combinatorial screening approach of cells derived from three pediatric tumor types, we identified Ewing sarcoma-specific interactions of a diverse set of targeted agents including approved drugs. We were able to retrieve highly synergistic drug combinations specific for Ewing sarcoma and identified signaling processes important for Ewing sarcoma cell proliferation determined by EWS-FLI1 We generated a molecular target profile of PKC412, a multikinase inhibitor with strong synergistic propensity in Ewing sarcoma, revealing its targets in critical Ewing sarcoma signaling routes. Using a multilevel experimental approach including quantitative phosphoproteomics, we analyzed the molecular rationale behind the disease-specific synergistic effect of simultaneous application of PKC412 and IGF1R inhibitors. The mechanism of the drug synergy between these inhibitors is different from the sum of the mechanisms of the single agents. The combination effectively inhibited pathway crosstalk and averted feedback loop repression, in EWS-FLI1-dependent manner. Mol Cancer Ther; 16(1); 88-101. ©2016 AACR.


Redundant Innate and Adaptive Sources of IL17 Production Drive Colon Tumorigenesis.
IL17-producing Th17 cells, generated through a STAT3-dependent mechanism, have been shown to promote carcinogenesis in many systems, including microbe-driven colon cancer. Additional sources of IL17, such as γδ T cells, become available under inflammatory conditions, but their contributions to cancer development are unclear. In this study, we modeled Th17-driven colon tumorigenesis by colonizing Min(Ap) (c+/-) mice with the human gut bacterium, enterotoxigenic Bacteroides fragilis (ETBF), to investigate the link between inflammation and colorectal cancer. We found that ablating Th17 cells by knocking out Stat3 in CD4(+) T cells delayed tumorigenesis, but failed to suppress the eventual formation of colonic tumors. However, IL17 blockade significantly attenuated tumor formation, indicating a critical requirement for IL17 in tumorigenesis, but from a source other than Th17 cells. Notably, genetic ablation of γδ T cells in ETBF-colonized Th17-deficient Min mice prevented the late emergence of colonic tumors. Taken together, these findings support a redundant role for adaptive Th17 cell- and innate γδT17 cell-derived IL17 in bacteria-induced colon carcinogenesis, stressing the importance of therapeutically targeting the cytokine itself rather than its cellular sources. Cancer Res; 76(8); 2115-24. ©2016 AACR.


Radiofrequency ablation of osteoid osteoma.
Osteoid osteoma is a benign bone neoplasm with a reported incidence of 2-3% among all bone primary tumors. Although it is a small and benign lesion, it is often cause of patient complaint and discomfort. It is generally characterized by a long lasting, unremitting pain that typically exacerbates at night, often leading to sleep deprivation and functional limitation of the skeletal segment involved, with a significant reduction of patient daily life activities and consequent worsening of the overall quality of life. Over decades, complete surgical resection has represented the only curative treatment for symptomatic patients. In the last years, new percutaneous ablation techniques, especially radiofrequency ablation, have been reported to be a safe and effective alternative to classical surgery, with a low complication and recurrence rate, and a significant reduction in hospitalization cost and duration. The aim of this article is to provide an overview about the radiofrequency thermal ablation procedure in the treatment of osteoid osteoma.


Investigation of UV optical fibers under synchrotron irradiation.
The focus of the present paper deals, for the first time, with commercial UV optical fibers, characterizing their behaviour as they are subjected to very high flux wiggler generated synchrotron radiation. Five distinct types of UV optical fibers, produced by three manufactures, were exposed to total doses between 5 Gy and 2000 Gy. The exposure to synchrotron radiation was performed in two campaigns. The tests were run off-line and considered the dependence of the radiation induced attenuation (RIA) as function of the total dose. The recovery of the radiation induced colour centres was studied at room temperature and after heating the samples up to 560 K. As a première, we also investigated through THz imaging and spectroscopy the irradiated optical fiber samples. Under these conditions, three of the optical fibers proved to be radiation resistant. The two optical fibers sensitive to synchrotron radiation exhibited a linear variation of the optical absorption at the wavelengths of λ = 229 nm, λ = 248 nm, and λ = 265 nm, for total doses between 60 Gy and 2000 Gy. These two samples showed also an increase of the optical absorption in the UV spectral range when heated to 560 K. The optical fibers sensitive to synchrotron radiation can potentially be used for on-line radiation dosimetry.


Population biobanking and international collaboration.
Population-based biobanks promise to be important resources for genetic research. However, the study of normal genomic variation across populations requires the collection of data and biological samples from individuals on a large scale. While international collaboration has become both a scientific and an ethical imperative, international sharing of data and samples poses many challenges. Significant variation persists among the legal and ethical norms governing population biobanks in different jurisdictions. Many of these norms do not clearly provide for international access. To illustrate these problems, we collected and compared applicable legislative instruments, as well as ethical guidelines issued by national, regional, and international bodies. In addition, harmonization is faced with important limitations and may not be sufficient to ensure effective international sharing. Population biobanks are therefore looking for new ways to promote sharing and improve interoperability. The formation of biobank networks and the development of common governance tools are two approaches that are setting the groundwork for international collaboration in genetic research.


EphA4 signaling regulates phospholipase Cgamma1 activation, cofilin membrane association, and dendritic spine morphology.
Specialized postsynaptic structures known as dendritic spines are the primary sites of glutamatergic innervation at synapses of the CNS. Previous studies have shown that spines rapidly remodel their actin cytoskeleton to modify their shape and this has been associated with changes in synaptic physiology. However, the receptors and signaling intermediates that restructure the actin network in spines are only beginning to be identified. We reported previously that the EphA4 receptor tyrosine kinase regulates spine morphology. However, the signaling pathways downstream of EphA4 that induce spine retraction on ephrin ligand binding remain poorly understood. Here, we demonstrate that ephrin stimulation of EphA4 leads to the recruitment and activation of phospholipase Cgamma1 (PLCgamma1) in heterologous cells and in hippocampal slices. This interaction occurs through an Src homology 2 domain of PLCgamma1 and requires the EphA4 juxtamembrane tyrosines. In the brain, PLCgamma1 is found in multiple compartments of synaptosomes and is readily found in postsynaptic density fractions. Consistent with this, PLC activity is required for the maintenance of spine morphology and ephrin-induced spine retraction. Remarkably, EphA4 and PLC activity modulate the association of the actin depolymerizing/severing factor cofilin with the plasma membrane. Because cofilin has been implicated previously in the structural plasticity of spines, this signaling may enable cofilin to depolymerize actin filaments and restructure spines at sites of ephrin-EphA4 contact.


Gangliosides induce selective modulation of CD4 from helper T lymphocytes.
The cluster designation (CD)4 molecule is one of several nonpolymorphic T lymphocyte surface proteins that have been implicated in T cell-target cell interactions, and is thought to play an important role in regulating T helper cell function. Previously, we found that gangliosides inhibited the function of rat T helper cell lines, and simultaneously inhibited the expression of the rat CD4 molecule identified by the W3/25 antibody. We have now evaluated the generality and mechanism(s) of ganglioside-induced modulation of CD4 expressed by mouse, rat, and human T helper lymphocytes. Ganglioside pretreatment induced rapid and selective disappearance of the CD4 molecule from T helper cells of all three species. The ganglioside effect was temperature- and dose-dependent, reversible within 24 hr of ganglioside removal, azide-insensitive, and was neutralized completely by 10% serum. CD4 modulation appeared to be a general property of gangliosides since the effect could be induced similarly by highly purified individual gangliosides with varying amounts of sialic acid, or by mixed gangliosides. The activity of gangliosides appeared to require both the lipid and sialated oligosaccharide moieties. Gangliosides did not inactivate antibody function, but prevented binding at the cell surface by 12 different monoclonal antibodies specific for a variety of different CD4 epitopes. Preclearance of CD4 by antibody-mediated capping reduced binding of 3H-GM1 to T helper cells. Labeled GM1 bound to several detergent-extracted and transblotted lymphocyte-associated proteins, but apparently did not bind directly to the CD4 molecule under these conditions. These results indicate that gangliosides induce a profound change in the molecular orientation of CD4 within the T helper cell membrane which renders epitopes on the CD4 molecule inaccessible to antibody. This ganglioside effect represents a novel pathway which may contribute to the understanding of the role of CD4 as a regulatory molecule and as a specific receptor for the acquired immune deficiency syndrome virus.


Hedgehog acts by distinct gradient and signal relay mechanisms to organise cell type and cell polarity in the Drosophila abdomen.
The epidermis of the adult Drosophila abdomen is formed by a chain of anterior (A) and posterior (P) compartments, each segment comprising one A and one P compartment. In the accompanying paper (Struhl et al., 1997), we provide evidence that Hedgehog protein (Hh), being secreted from P compartment cells, organises the pattern and polarity of A compartment cells. Here we test whether Hh acts directly or by a signal relay mechanism. We use mutations in Protein Kinase A (PKA) or smoothened (smo) to activate or to block Hh signal transduction in clones of A compartment cells. For cell type, a scalar property, both manipulations cause strictly autonomous transformations: the cells affected are exactly those and only those that are mutant. Hence, we infer that Hh acts directly on A compartment cells to specify the various types of cuticular structures that they differentiate. By contrast, these same manipulations cause non-autonomous effects on cell polarity, a vectorial property. Consequently, we surmise that Hh influences cell polarity indirectly, possibly by inducing other signalling factors. Finally, we present evidence that Hh does not polarise abdominal cells by utilising either Decapentaplegic (Dpp) or Wingless (Wg), the two morphogens through which Hh acts during limb development. We conclude that, in the abdomen, cell type and cell polarity reflect distinct outputs of Hh signalling and propose that these outputs are controlled by separable gradient and signal relay mechanisms.


Impulsivity resulting from frontostriatal dysfunction in drug abuse: implications for the control of behavior by reward-related stimuli.
Drug abuse and dependence define behavioral states involving increased allocation of behavior towards drug seeking and taking at the expense of more appropriate behavioral patterns. As such, addiction can be viewed as increased control of behavior by the desired drug (due to its unconditioned, rewarding properties). It is also clear that drug-associated (conditioned) stimuli acquire heightened abilities to control behaviors. These phenomena have been linked with dopamine function within the ventral striatum and amygdala and have been described specifically in terms of motivational and incentive learning processes. New data are emerging that suggest that regions of the frontal cortex involved in inhibitory response control are directly affected by long-term exposure to drugs of abuse. The result of chronic drug use may be frontal cortical cognitive dysfunction, resulting in an inability to inhibit inappropriate unconditioned or conditioned responses elicited by drugs, by related stimuli or by internal drive states. Drug-seeking behavior may thus be due to two related phenomena: (1) augmented incentive motivational qualities of the drug and associated stimuli (due to limbic/amygdalar dysfunction) and (2) impaired inhibitory control (due to frontal cortical dysfunction). In this review, we consider the neuro-anatomical and neurochemical substrates subserving inhibitory control and motivational processes in the rodent and primate brain and their putative impact on drug seeking. The evidence for cognitive impulsivity in drug abuse associated with dysfunction of the frontostriatal system will be discussed, and an integrative hypothesis for compulsive reward-seeking in drug abuse will be presented.


How 6 Organizations Developed Tools and Processes for Social Determinants of Health Screening in Primary Care: An Overview.
Little is known about how health care organizations are developing tools for identifying/addressing patients' social determinants of health (SDH). We describe the processes recently used by 6 organizations to develop SDH screening tools for ambulatory care and the barriers they faced during those efforts. Common processes included reviewing literature and consulting primary care staff. The organizations prioritized avoiding redundant data collection, integrating SDH screening into existing workflows, and addressing diverse clinic needs. This article provides suggestions for others hoping to develop similar tools/strategies for identifying patients' SDH needs in ambulatory care settings, with recommendations for further research.


Medicinal and cosmetic uses of Bee's Honey - A review.
Bee's honey is one of the most valued and appreciated natural substances known to mankind since ancient times. There are many types of bee's honey mentioned in Ayurveda. Their effects differ and 'Makshika' is considered medicinally the best. According to modern scientific view, the best bee's honey is made by Apis mellifera (Family: Apidae). In Sri Lanka, the predominant honey-maker bee is Apis cerana. The aim of this survey is to emphasize the importance of bee's honey and its multitude of medicinal, cosmetic and general values. Synonyms, details of formation, constitution, properties, and method of extraction and the usages of bee's honey are gathered from text books, traditional and Ayurvedic physicians of Western and Southern provinces, villagers of 'Kalahe' in Galle district of Sri Lanka and from few search engines. Fresh bee's honey is used in treatment of eye diseases, throat infections, bronchial asthma, tuberculosis, hiccups, thirst, dizziness, fatigue, hepatitis, worm infestation, constipation, piles, eczema, healing of wounds, ulcers and used as a nutritious, easily digestible food for weak people. It promotes semen, mental health and used in cosmetic purposes. Old bee's honey is used to treat vomiting, diarrhea, rheumatoid arthritis, obesity, diabetes mellitus and in preserving meat and fruits. Highly popular in cosmetic treatment, bee's honey is used in preparing facial washes, skin moisturizers, hair conditioners and in treatment of pimples. Bee's honey could be considered as one of the finest products of nature that has a wide range of beneficial uses.


Multiple lineages of ancient CR1 retroposons shaped the early genome evolution of amniotes.
Chicken repeat 1 (CR1) retroposons are long interspersed elements (LINEs) that are ubiquitous within amniote genomes and constitute the most abundant family of transposed elements in birds, crocodilians, turtles, and snakes. They are also present in mammalian genomes, where they reside as numerous relics of ancient retroposition events. Yet, despite their relevance for understanding amniote genome evolution, the diversity and evolution of CR1 elements has never been studied on an amniote-wide level. We reconstruct the temporal and quantitative activity of CR1 subfamilies via presence/absence analyses across crocodilian phylogeny and comparative analyses of 12 crocodilian genomes, revealing relative genomic stasis of retroposition during genome evolution of extant Crocodylia. Our large-scale phylogenetic analysis of amniote CR1 subfamilies suggests the presence of at least seven ancient CR1 lineages in the amniote ancestor; and amniote-wide analyses of CR1 successions and quantities reveal differential retention (presence of ancient relics or recent activity) of these CR1 lineages across amniote genome evolution. Interestingly, birds and lepidosaurs retained the fewest ancient CR1 lineages among amniotes and also exhibit smaller genome sizes. Our study is the first to analyze CR1 evolution in a genome-wide and amniote-wide context and the data strongly suggest that the ancestral amniote genome contained myriad CR1 elements from multiple ancient lineages, and remnants of these are still detectable in the relatively stable genomes of crocodilians and turtles. Early mammalian genome evolution was thus characterized by a drastic shift from CR1 prevalence to dominance and hyperactivity of L2 LINEs in monotremes and L1 LINEs in therians.


Properties of resistant cells generated from lung cancer cell lines treated with EGFR inhibitors.
Epidermal growth factor receptor (EGFR) signaling plays an important role in non-small cell lung cancer (NSCLC) and therapeutics targeted against EGFR have been effective in treating a subset of patients bearing somatic EFGR mutations. However, the cancer eventually progresses during treatment with EGFR inhibitors, even in the patients who respond to these drugs initially. Recent studies have identified that the acquisition of resistance in approximately 50% of cases is due to generation of a secondary mutation (T790M) in the EGFR kinase domain. In about 20% of the cases, resistance is associated with the amplification of MET kinase. In the remaining 30-40% of the cases, the mechanism underpinning the therapeutic resistance is unknown. An erlotinib resistant subline (H1650-ER1) was generated upon continuous exposure of NSCLC cell line NCI-H1650 to erlotinib. Cancer stem cell like traits including expression of stem cell markers, enhanced ability to self-renew and differentiate, and increased tumorigenicity in vitro were assessed in erlotinib resistant H1650-ER1 cells. The erlotinib resistant subline contained a population of cells with properties similar to cancer stem cells. These cells were found to be less sensitive towards erlotinib treatment as measured by cell proliferation and generation of tumor spheres in the presence of erlotinib. Our findings suggest that in cases of NSCLC accompanied by mutant EGFR, treatment targeting inhibition of EGFR kinase activity in differentiated cancer cells may generate a population of cancer cells with stem cell properties.


Effect of orthostatic hypotension on sustained attention in patients with autonomic failure.
Orthostatic hypotension has been associated with impaired cognitive function, but cognitive function during orthostatic hypotension has hardly been studied. We studied the effect of orthostatic hypotension, induced by head-up tilt (HUT), on sustained attention in patients with autonomic failure. We studied the sustained attention to response task (SART) in the supine position and during HUT in 10 patients with autonomic failure and 10 age-matched and sex-matched controls. To avoid syncope, the tilting angle was tailored to patients to reach a stable systolic blood pressure below 100 mm Hg. Controls were all tilted at an angle of 60°. Cerebral blood flow velocity, blood pressure and heart rate were measured continuously. In patients, systolic blood pressure was 61.4 mm Hg lower during HUT than in the supine position (p<0.001). Patients did not make more SART errors during HUT than in the supine position (-1.3 errors, p=0.3). Controls made 2.3 fewer errors during SART in the HUT position compared to the supine position (p=0.020). SART performance led to an increase in systolic blood pressure (+11.8 mm Hg, p=0.018) and diastolic blood pressure (+5.8 mm Hg, p=0.017) during SART in the HUT position, as well as to a trend towards increased cerebral blood flow velocity (+3.8 m/s, p=0.101). Orthostatic hypotension in patients with autonomic failure was not associated with impaired sustained attention. This might partly be explained by the observation that SART performance led to a blood pressure increase. Moreover, the upright position was associated with better performance in controls and, to a lesser extent, also in patients.


Reevaluation of Host Specificity of the Closely Related Species Pseudoperonospora humuli and P. cubensis.
Pseudoperonospora cubensis causes some of the most devastating diseases of cucurbitaceous crops, while P. humuli is an important pathogen of hop (Cannabaceae). Although parasitic to different Angiosperm orders, these pathogens are highly similar, both in morphology and based on molecular comparisons. Considering the close relationship of P. humuli and P. cubensis, it was hypothesized that cross infectivity of the pathogens between their optimum hosts might be possible. Two strains of P. humuli and one of P. cubensis used in this study were able to reproduce on the two wild cucurbit relatives Bryonia dioica and Sicyos angulatus. Interestingly, limited infectivity of the P. cubensis strain to hop was also observed, and the P. humuli strain was also able to infest Cucumis sativus. The cross infections were verified by comparing the nuclear ribosomal-internal transcribed spacer sequences of newly produced sporangiophores from the different hosts. Morphologically the two pathogens could be distinguished by statistical analyses on their original hosts but these differences diminished on some alternate hosts. The ability of P. cubensis to infest the perennial Bryonia dioica and Humulus lupulus might offer a limited possibility for overwintering in temperate regions and needs to be evaluated in future studies. The ability of P. humuli to complete its life cycle on all the tested cucurbits provides evidence that the host jump across orders may continue and points to the high virulence potential of this pathogen.


Selected cannabis terpenes synergize with THC to produce increased CB1 receptor activation.
The cannabis plant exerts its pharmaceutical activity primarily by the binding of cannabinoids to two G protein-coupled cannabinoid receptors, CB1 and CB2. The role that cannabis terpenes play in this activation has been considered and debated repeatedly, based on only limited experimental results. In the current study we used a controlled in-vitro heterologous expression system to quantify the activation of CB1 receptors by sixteen cannabis terpenes individually, by tetrahydrocannabinol (THC) alone and by THC-terpenes mixtures. The results demonstrate that all terpenes, when tested individually, activate CB1 receptors, at about 10-50% of the activation by THC alone. The combination of some of these terpenes with THC significantly increases the activity of the CB1 receptor, compared to THC alone. In some cases, several fold. Importantly, this amplification is evident at terpene to THC ratios similar to those in the cannabis plant, which reflect very low terpene concentrations. For some terpenes, the activation obtained by THC- terpene mixtures is notably greater than the sum of the activations by the individual components, suggesting a synergistic effect. Our results strongly support a modulatory effect of some of the terpenes on the interaction between THC and the CB1 receptor. As the most effective terpenes are not necessarily the most abundant ones in the cannabis plant, reaching "whole plant" or "full spectrum" composition is not necessarily an advantage. For enhanced therapeutic effects, desired compositions are attainable by enriching extracts with selected terpenes. These compositions adjust the treatment for various desired medicinal and personal needs.


Concordant evolution of plumage colour, feather microstructure and a melanocortin receptor gene between mainland and island populations of a fairy-wren.
Studies of the patterns of diversification of birds on islands have contributed a great deal to the development of evolutionary theory. In white-winged fairy-wrens, Malurus leucopterus, mainland males develop a striking blue nuptial plumage whereas those on nearby islands develop black nuptial plumage. We explore the proximate basis for this divergence by combining microstructural feather analysis with an investigation of genetic variation at the melanocortin-1 receptor locus (MC1R). Fourier analysis revealed that the medullary keratin matrix (spongy layer) of the feather barbs of blue males was ordered at the appropriate nanoscale to produce the observed blue colour by coherent light scattering. Surprisingly, the feather barbs of black males also contained a spongy layer that could produce a similar blue colour. However, black males had more melanin in their barbs than blue males, and this melanin may effectively mask any structural colour produced by the spongy layer. Moreover, the presence of this spongy layer suggests that black island males evolved from a blue-plumaged ancestor. We also document concordant patterns of variation at the MC1R locus, as five amino acid substitutions were perfectly associated with the divergent blue and black plumage phenotypes. Thus, with the possible involvement of a melanocortin receptor locus, increased melanin density may mask the blue-producing microstructure in black island males, resulting in the divergence of plumage coloration between mainland and island white-winged fairy-wrens. Such mechanisms may also be responsible for plumage colour diversity across broader geographical and evolutionary scales.


Protein-protein interactions in paralogues: Electrostatics modulates specificity on a conserved steric scaffold.
An improved knowledge of protein-protein interactions is essential for better understanding of metabolic and signaling networks, and cellular function. Progress tends to be based on structure determination and predictions using known structures, along with computational methods based on evolutionary information or detailed atomistic descriptions. We hypothesized that for the case of interactions across a common interface, between proteins from a pair of paralogue families or within a family of paralogues, a relatively simple interface description could distinguish between binding and non-binding pairs. Using binding data for several systems, and large-scale comparative modeling based on known template complex structures, it is found that charge-charge interactions (for groups bearing net charge) are generally a better discriminant than buried non-polar surface. This is particularly the case for paralogue families that are less divergent, with more reliable comparative modeling. We suggest that electrostatic interactions are major determinants of specificity in such systems, an observation that could be used to predict binding partners.


ICTV Virus Taxonomy Profile: Bromoviridae.
Bromoviridae is a family of plant viruses with tri-segmented, positive-sense, single-stranded RNA genomes of about 8 kb in total. Genomic RNAs are packaged in separate virions that may also contain subgenomic, defective or satellite RNAs. Virions are variable in morphology (spherical or bacilliform) and are transmitted between hosts mechanically, in/on the pollen and non-persistently by insect vectors. Members of the family are responsible for major disease epidemics in fruit, vegetable and fodder crops such as tomato, cucurbits, bananas, fruit trees and alfalfa. This is a summary of the International Committee on Taxonomy of Viruses (ICTV) Report on the family Bromoviridae, which is available at www.ictv.global/report/bromoviridae.


Validity of actigraphs uniaxial and triaxial accelerometers for assessment of physical activity in adults in laboratory conditions.
Few studies to date have directly compared the Actigraphs GT1M and the GT3X, it would be of tremendous value to know if these accelerometers give similar information about intensities of PA. Knowing if output is similar would have implications for cross-examination of studies. The purpose of the study was to assess the validity of the GT1M and the GT3X Actigraph accelerometers for the assessment of physical activity against oxygen consumption in laboratory conditions. Forty-two college-aged participants aged 18-25 years wore the GT1M and the GT3X on their right hip during treadmill exercise at three different speeds, slow walking 4.8 km.h-1, fast walking 6.4 km.h-1, and running 9.7 km.h-1). Oxygen consumption was measured minute-by minute using a metabolic system. Bland-Altman plots were used to assess agreement between activity counts from the GT3X and GT1M, and correlations were assessed the ability of the accelerometers to assess physical activity. Bias for 4.8 km.h-1 was 2814.4 cpm (limits 1211.3 to 4417.4), for 6.4 km.h-1 was 3713.6 cpm (limits 1573.2 to 5854.0), and for 9.7 km.h-1 was-3811.2 cpm (limits 842.1 to 6780.3). Correlations between counts per minute for the GT1M and the GT3X were significantly correlated with VO2 (r = 0.881, p < 0.001; r = 0.810, p < 0.001 respectively). The present study showed that both the GT1M and the GT3X accurately measure physical activity when compared to oxygen consumption.


Enhancement of radiation and chemotherapeutic drug responses by 2-deoxy-D-glucose in animal tumors.
The development of an approach based on the energy-linked modification of DNA repair and cellular recovery processes using 2-deoxy-D-glucose (2-DG; inhibitor of glycolytic ATP production) has shown promising results in a number of model systems of cancer. Following encouraging results on the tolerance and toxicity (acute as well as late effects) of the combination (2-DG and hypofractionated radiotherapy) in Phase I and II clinical trials, its efficacy is currently under evaluation in Phase III clinical trials for glioma patients. Since heterogeneous physiologic and metabolic status in tumors as well as host-tumor interactions influence the local tumor control, which coupled with systemic disturbances could determine the cure (long-term tumor free survival), investigations on the in vivo responses of tumors to the combined treatment have received considerable attention. This communication provides a brief overview on the in vivo studies related to radio- and chemosensitization of tumors by 2-DG, besides the normal tissue toxicity induced by the combined treatment of 2-DG and radiation or chemotherapeutic drugs.


Nalbuphine for postoperative pain treatment in children.
Several surveys over the past few years have demonstrated that postoperative pain in children is not treated appropriately. One pharmacological treatment option in a multimodal approach for postoperative pain treatment is the systemic administration of opioids. However, opioids are rarely used for postoperative pain treatment in children due to fear of adverse events. One long-standing opioid for systemic use is nalbuphine, a kappa-receptor agonist and µ-receptor antagonist. The efficacy of nalbuphine is believed to be similar to morphine. Increased dosing might result in a ceiling effect, and thus less analgesia than expected. In addition, there might be a lower risk for opioid-induced side effects (nausea, vomiting) and severe adverse events (respiratory depression) due to the antagonistic effect of the µ-receptor. Nalbuphine may be an useful opioid for postoperative use in children, but exact efficacy (e.g. compared to other commonly used opioids) has not been determined yet. To assess the efficacy and adverse events of nalbuphine for acute postoperative pain treatment in children undergoing surgery. We systematically searched the following databases: The Cochrane Central Register of Controlled Trials (CENTRAL) (The Cochrane Library 2013, Issue 7), MEDLINE via Pubmed (January 1966 to July 2013) and EMBASE via Ovid (January 1947 to July 2013). We did not impose any restrictions regarding language or publication date. We checked all reference lists of retrieved articles for additional references. All randomised controlled trials (RCTs) investigating nalbuphine compared with placebo or other opioids were included. Two review authors independently scanned the retrieved articles and made a decision regarding inclusion or exclusion of studies for this review. The same authors also performed the data extraction and the assessment of risk of bias. Ten RCTs including 658 patients were finally included in this systematic review. Five trials compared nalbuphine with placebo. Data from one out of five studies for the outcome moderate/severe pain following nalbuphine compared to placebo gave a risk ratio (RR) 1 hour postoperatively (postop) of 0.1 (95% confidence interval (CI) 0.01 to 0.71; low quality evidence) and a RR 2 hours postop of 0.14 (95% CI 0.02 to 1.06; low quality evidence). The estimated RR based on data from a single study indicated that nalbuphine reduced the requirement for analgesia two hours postop (RR 0.47; 95% CI 0.27 to 0.84; low quality evidence). Two included trials compared nalbuphine with morphine and showed a nonsignificant lower or comparable RR for moderate/severe pain at 1 hour postop (RR 0.84; 95% CI 0.12 to 5.74; low quality evidence), and 2 hours postop (RR 1.09; 95% CI 0.59 to 2.01; low quality evidence) for nalbuphine versus morphine. Four trials compared nalbuphine with tramadol for postoperative pain; data from one trial (per outcome) revealed a lower but nonsignificant RR for the need of additional rescue analgesics in children receiving nalbuphine (RR 2 hours postop 0.75; 95% CI 0.39 to 1.43; low quality evidence) (RR 12 hours postop 0.33; 95% CI 0.04 to 2.77; low quality evidence). One out of three trials comparing nalbuphine with pethidine demonstrated that the RR was not significantly lower following nalbuphine administration compared to pethidine (RR 2 hours postop 1.07; 95% CI 0.52 to 2.23; low quality evidence) (RR 24 hours postop 1.13; 95% CI 0.52 to 2.44; very low quality evidence). The most common adverse event was postoperative nausea and vomiting (PONV). Only one included trial reported that the RR for PONV in the postoperative care unit (PACU) was not significantly higher following nalbuphine compared to placebo (RR 1.00; 95% CI 0.16 to 6.42; low quality evidence) nor to morphine (RR 1.33; 95% CI 0.64 to 2.77; low quality evidence). Because the overall quality of available evidence was low, this systematic review could not definitively show that the analgesic efficacy of nalbuphine is superior compared to placebo. Furthermore, due to the lack of significant results the comparison with other common opioids is also unclear. The same holds true for the evidence focusing on adverse events following nalbuphine compared to placebo or other opioid administration. The evidence is limited, because studies did not report conclusively all important postoperative pain outcomes (e.g. number of patients with the need for rescue analgesia, postoperative pain scores). Thus, a quantitative analysis was not possible for many major aspects (e.g. rescue analgesia, pain scores) and heterogeneity could not be further explored.


Role of midbrain in the control of breathing in anuran amphibians.
The present study was designed to explore systematically the midbrain of unanesthetized, decerebrate anuran amphibians (bullfrogs), using chemical and electrical stimulation and midbrain transections to identify sites capable of exciting and inhibiting breathing. Ventilation was measured as fictive motor output from the mandibular branch of the trigeminal nerve and the laryngeal branch of the vagus nerve. The results of our transection studies suggest that, under resting conditions, the net effect of inputs from sites within the rostral half of the midbrain is to increase fictive breathing frequency, whereas inputs from sites within the caudal half of the midbrain have no net effect on fictive breathing frequency but appear to act on the medullary central rhythm generator to produce episodic breathing. The results of our stimulation experiments indicate that the principal sites in the midbrain that are capable of exciting or inhibiting the fictive frequency of lung ventilation, and potentially clustering breaths into episodes, appear to be those primarily involved in visual and auditory integration, motor functions, and attentional state.


California Liaison Committee for Podiatric Medical Education and Training.
Podiatric medicine faces some significant challenges that threaten its future growth and development. The California Liaison Committee for Podiatric Medical Education and Training exemplifies an innovative approach to meeting these challenges. The California Liaison Committee has established a dialogue among California's college-based and community-based podiatric medical educators, licensing board members, and private practitioners. The work of the California Liaison Committee, unprecedented in the state, effectively facilitates the curriculum transformation process through cooperation and collaboration.


Impact of a web based interactive simulation game (PULSE) on nursing students' experience and performance in life support training--a pilot study.
The delivery of effective life support measures is highly associated with the quality, design and implementation of the education that underpins it. Effectively responding to a critical event is a requirement for all nurses illustrating the need for effective educational approaches from pre-registration training through to enhancing and maintaining life support skills after qualification. This paper reports the findings of utilising a web-based multimedia simulation game PULSE (Platform for Undergraduate Life Support Education). The platform was developed to enhance the student experience of life support education, to motivate on-going learning and engagement and to improve psychomotor skills associated with the provision of Intermediate Life Support (ILS) training. Pre training participants played PULSE and during life support training data was collected from an intervention and a control group of final year undergraduate nursing students (N=34). Quantitative analysis of performance took place and qualitative data was generated from a questionnaire assessing the learning experience. A statistically significant difference was found between the competence the groups displayed in the three skills sets of checking equipment, airway assessment and the safe/effective use of defibrillator at ILS level, and PULSE was positively evaluated as an educational tool when used alongside traditional life support training.


Mining the Genome of Streptomyces leeuwenhoekii: Two New Type I Baeyer-Villiger Monooxygenases From Atacama Desert.
Actinobacteria are an important source of commercial (bio)compounds for the biotechnological and pharmaceutical industry. They have also been successfully exploited in the search of novel biocatalysts. We set out to explore a recently identified actinomycete, Streptomyces leeuwenhoekii C34, isolated from a hyper-arid region, the Atacama desert, for Baeyer-Villiger monooxygenases (BVMOs). Such oxidative enzymes are known for their broad applicability as biocatalysts by being able to perform various chemical reactions with high chemo-, regio-, and/or enantioselectivity. By choosing this specific Actinobacterium, which comes from an extreme environment, the respective enzymes are also expected to display attractive features by tolerating harsh conditions. In this work, we identified two genes in the genome of S. leeuwenhoekii (sle_13190 and sle_62070) that were predicted to encode for Type I BVMOs, the respective flavoproteins share 49% sequence identity. The two genes were cloned, overexpressed in E. coli with phosphite dehydrogenase (PTDH) as fusion partner and successfully purified. Both flavin-containing proteins showed NADPH-dependent Baeyer-Villiger oxidation activity for various ketones and sulfoxidation activity with some sulfides. Gratifyingly, both enzymes were found to be rather robust by displaying a relatively high apparent melting temperature (45°C) and tolerating water-miscible cosolvents. Specifically, Sle_62070 was found to be highly active with cyclic ketones and displayed a high regioselectivity by producing only one lactone from 2-phenylcyclohexanone, and high enantioselectivity by producing only normal (-)-1S,5R and abnormal (-)-1R,5S lactones (ee > 99%) from bicyclo[3.2.0]hept-2-en-6-one. These two newly discovered BVMOs add two new potent biocatalysts to the known collection of BVMOs.


Molecular genetic profiles among individual Clonorchis sinensis adults collected from cats in two geographic regions of China revealed by RAPD and MGE-PCR methods.
Clonorchis sinensis causes the important food-borne zoonosis, clonorchiasis, which is endemic in East Asia, especially in China mainly in Guangdong, Guangxi and Heilongjiang provinces and Korea. Although comparisons on isoenzymes and some molecular profiles of C. sinensis collected from different parts of China and Korea have been studied, few works on the genetic variation among the individuals from different regions of China has been reported. In the present study, individual adults of C. sinensis were collected from cats in two geographic locations (Guangdong province in the South and Heilongjiang province in the North) of China and 44 of them were examined by using random amplified polymorphic DNA (RAPD)-PCR and mobile genetic elements (MGEs)-PCR techniques to assess the individual genetic variability within and between the two groups of this parasite. Six arbitrary primers and two pairs of MGE primers were employed in the genomic DNA amplification. The molecular patterns showed significant polymorphism among the individuals. The RAPD data displayed that the similarity coefficient (SC) of the individuals within Heilongjiang group was much higher than that of the Guangdong group, which was further confirmed by MGE-PCR results. Individuals from Heilongjiang were found genetically closer with lesser polymorphisms than those collected from Guangdong province. These results demonstrated that RAPD and MGE-PCR techniques, particularly RAPD method, could be useful for investigating genetic variations among C. sinensis individuals. They may also indicate that the genetic variation of C. sinensis occurs in the subtropical region--Guangdong--faster than that in the cold-region--Heilongjiang province--due to more generations (life cycle) occurred.


Temporal patterns and space-time cluster analysis of foot-and-mouth disease (FMD) cases from 2007 to 2017 in Vietnam.
In Vietnam, Foot-and-mouth disease (FMD) is endemic, but no nationwide studies have been conducted to assess the monthly variations and space-time clusters of FMD. The main objective was to identify the temporal patterns and space-time clusters of FMD from 2007 to 2017 using national surveillance data in Vietnam. A total of 163,733 cases were reported from 2007 to 2017. Among them, the proportion of buffaloes (43.31% of total reported cases; 70,909 cases) was highest followed by cattle (30.11%; 49,306 cases), pigs (26.67%; 43,662 cases) and sheep/goats (0.41%; 675 cases). The serotype O was widely distributed across the country while serotype A was observed in Northeast, Central and Southern part of Vietnam while Asia 1 has been not identified since 2007. For monthly variations, most cases were observed during the dry season (from November to March) except Central Highlands. Under the spatial window was set at 50%, a total of seven clusters were identified. The primary cluster was observed from Dec 2009 to Dec 2010 in the northwest (radius: 101.67 km), showing a ratio of 3.75. The secondary cluster was detected in the northeast region (radius: 76.54 km) with a ratio of 3.53 in Feb 2017. The 3rd cluster was the largest with a radius of 176.69 km and located in the southern part of Vietnam. Interestingly, the most temporal clusters included between December and March during the study period. Our findings provide better insight into the temporal patterns and distribution of clusters of FMD in Vietnam. This study provides useful information to policymakers on the hotspot areas and timing of outbreaks. It also identifies when and where national surveillance and control programmes could be implemented more efficiently for the prevention and control of FMD.


Biochanin-A induces apoptosis and suppresses migration in FaDu human pharynx squamous carcinoma cells.
The aim of the present study was to investigate biochanin-A-induced anticancer effects and their cellular signaling pathway in FaDu pharyngeal squamous carcinoma cells. Biochanin-A induced cell death through increased cytotoxicity of FaDu cells in a dose- and time-dependent manner. The number of cells with nucleus condensation and the apoptotic population were increased in the FaDu cells stimulated with biochanin-A for 24 h. Furthermore, extrinsic apoptotic factors such as FasL and their downstream target caspase-8 were increased and activated in the FaDu cells treated with biochanin-A in a dose-dependent manner. Moreover, biochanin-A decreased the expression of intrinsic anti-apoptotic factors such as Bcl-2 and Bcl-xL, and increased the level and activation of intrinsic apoptotic factors such as Bad and caspase-9. Finally, biochanin-A induced the activation of caspase-3 and Poly(ADP ribose) polymerase (PARP) in FaDu cells. Our results suggest that biochanin-A-induced apoptosis was mediated by death receptor mediated-extrinsic and mitochondria-dependent intrinsic apoptotic signaling pathways. Biochanin-A also inhibited wound healing migration and proliferation of FaDu cells via the downregulation and inactivation of matrix metalloproteinase-2 and -9 that are mediated by the suppression of p38, mitogen activated protein kinase (MAPK), NF-κB and Akt cellular signaling pathways. Therefore, these data suggest that the biochanin-A may act as a potential chemotherapeutic compound to treat head and neck cancer.