Solvent effects on the conversion of dicopper(II) micro-eta(2):eta(2)-peroxo to bis-micro-oxo dicopper(III) complexes: direct probing of the solvent interaction.
A new tridentate ligand, PYAN, is employed to investigate solvent influences for dioxygen reactivity with [Cu(PYAN)(MeCN)]B(C(6)F(5))(4) (1). Stopped-flow kinetic studies confirm that the adducts [[u(II)(PYAN)]2)(O(2))][B(C(6)F(5))(4)](2) (2(Peroxo)) and [[u(III)(PYAN)]2)(O)(2)][B(C(6)F(5))(4)](2) (2(Oxo)) are in rapid equilibrium. Thermodynamic parameters for the equilibrium between 2(Peroxo) and 2(Oxo) re as follows: THF, deltaH degrees approximately -15.7 kJ/mol, deltaS degrees approximately -83 J/K.mol; acetone, deltaH degrees approximately -15.8 kJ/mol, deltaS degrees approximately -76 J/K.mol. UV-visible absorption and resonance Raman spectroscopic signatures demonstrate that the equilibrium is highly solvent dependent; the mixture is mostly 2(Peroxo) in CH(2)Cl(2), but there are significantly increasing quantities of 2(Oxo) along the series methylene chloride --> diethyl ether --> acetone --> tetrahydrofuran (THF). Copper(II)-N(eq) stretches (239, 243, 244, and 246 cm(-)(1) in CH(2)Cl(2), Et(2)O, acetone, and THF, respectively) are identified for 2(Peroxo), but they are not seen in 2(Oxo), revealing for the first time direct evidence for solvent coordination in the more open 2(Peroxo) structure.


Cervical dystonia: disease profile and clinical management.
Cervical dystonia, the most common focal dystonia, frequently results in cervical pain and disability as well as impairments affecting postural control. The predominant treatment for cervical dystonia is provided by physicians, and treatment can vary from pharmacological to surgical. Little literature examining more conservative approaches, such as physical therapy, exists. This article reviews the etiology and pathophysiology of the disease as well as medical and physical therapist management for people with cervical dystonia.


Binding of lipid droplets and mitochondria to intermediate filaments in rat Leydig cells.
We have examined the distribution of lipid droplets and mitochondria in relation to the cytoskeletons of Leydig cells in primary culture by using light and electron microscopy on living, intact and detergent-extracted cells. After mild extraction with Triton X-100 lipid droplets and mitochondria retained their original distribution within the cell. Double immunofluorescent microscopy showed that both structures co-localise with intermediate filaments. Transmission electron microscopy of intact (unextracted) and mildly extracted Leydig cells showed that intermediate filaments are closely associated with mitochondria and lipid droplets. By examination of stereo pairs, intermediate filaments were shown to establish direct contact with mitochondria and lipid droplets. The association of droplets and mitochondria with intermediate filaments suggests possible mechanisms by which the transport of cholesterol takes place from droplets to mitochondria where this substrate enters the steroidogenic pathway.


Driving Barocaloric Effects in a Molecular Spin-Crossover Complex at Low Pressures.
Barocaloric effects─thermal changes in a material induced by applied hydrostatic pressure─offer promise for creating solid-state refrigerants as alternatives to conventional volatile refrigerants. To enable efficient and scalable barocaloric cooling, materials that undergo high-entropy, reversible phase transitions in the solid state in response to a small change in pressure are needed. Here, we report that pressure-induced spin-crossover (SCO) transitions in the molecular iron(II) complex Fe[HB(tz)3]2 (HB(tz)3- = bis[hydrotris(1,2,4-triazol-1-yl)borate]) drive giant and reversible barocaloric effects at easily accessible pressures. Specifically, high-pressure calorimetry and powder X-ray diffraction studies reveal that pressure shifts as low as 10 bar reversibly induce nonzero isothermal entropy changes, and a pressure shift of 150 bar reversibly induces a large isothermal entropy change (>90 J kg-1 K-1) and adiabatic temperature change (>2 K). Moreover, we demonstrate that the thermodynamics of the SCO transition can be fine-tuned through systematic deuteration of the tris(triazolyl)borate ligand. These results provide new insights into pressure-induced SCO transitions and further establish SCO materials as promising barocaloric materials.


Flexible Ultrathin Chip-Film Patch for Electronic Component Integration and Encapsulation using Atomic Layer-Deposited Al2O3-TiO2 Nanolaminates.
Plasma-enhanced atomic layer deposition (PEALD) is utilized to improve the barrier properties of an organic chip-film patch (CFP) when it is used as an implant to prevent moisture and ions from migrating into the embedded electronic circuits. For this purpose, surface condition and material properties of eight modifications of Al2O3-TiO2 nanolaminates sequentially deposited on polyimide PI-2611 films are evaluated in detail. The effect of stress-induced warpage of the deposited Al2O3-TiO2 on the wafer level is calculated with the Stoney equation and reveals higher tensile stress values while increasing the thickness of Al2O3-TiO2 nanolaminates from 20 up to 80 nm. Contact angle measurement and atomic force microscopy are used to investigate the surface energy and wettability, as well as the surface morphology of polyimide-Al2O3-TiO2 interfaces. We show that plasma treatment of pristine polyimide leads to an enhanced adhesion force of the PEAL-deposited layer by a factor of 1.3. The water vapor transmission rate (WVTR) is determined by exposing the coated polyimide films to 85% humidity and 23 °C and yields down to 1.58 × 10-3 g(H2O)/(m2 d). The data obtained are compared with alternative coating processes using the polymers parylene-C and benzocyclobutene (BCB). The latter shows higher WVTR values of 1.2 × 10-1 and 1.7 × 10-1 g(H2O)/(m2 d) compared to the PEALD-PI-2611 systems, indicating lower barrier properties. Two Al2O3-TiO2 modifications with low WVTR values have been chosen for encapsulating the CFP substrates and exposing them in a long-time experiment to chemical and mechanical loads in a chamber filled with phosphate-buffered saline at 37 °C, pH 7.3, and a cyclically applied pressure of 160 mbar (∼120 mm Hg). The electrical leakage behavior of the CFP systems is measured and reveals reliable electrical long-term stability far beyond 11 months, highlighting the great potential of PEALD-encapsulated CFPs.


Determination of concerted or stepwise mechanism of hydrogen tunneling from isotope effects: Departure between experiment and theory.
Isotope substitution is an important experimental technique that offers deep insight into reaction mechanisms, as the measured kinetic isotope effects (KIEs) can be directly compared with theory. For multiple proton transfer processes, there are two types of mechanisms: stepwise transfer and concerted transfer. The Bell-Limbach model provides a simple theory to determine whether the proton transfer mechanism is stepwise or concerted from KIEs. Recent scanning tunneling microscopy (STM) experiments have studied the proton switching process in water tetramers on NaCl(001). Theoretical studies predict that this process occurs via a concerted mechanism; however, the experimental KIEs resemble the Bell-Limbach model for stepwise tunneling, raising questions on the underlying mechanism or the validity of the model. We study this system using ab initio instanton theory, and in addition to thermal rates, we also considered microcanonical rates, as well as tunneling splittings. The instanton theory predicts a concerted mechanism, and the KIEs for tunneling rates (both thermal and microcanonical) upon deuteration are consistent with the Bell-Limbach model for concerted tunneling but could not explain the experiments. For tunneling splittings, partial and full deuteration change the size of it in a similar fashion to how they change the rates. We further examined the Bell-Limbach model in another system, porphycene, which has both stepwise and concerted tunneling pathways. The KIEs predicted by instanton theory are again consistent with the Bell-Limbach model. This study highlights differences between KIEs in stepwise and concerted tunneling and the discrepancy between theory and recent STM experiments. New theory/experiments are desired to settle this problem.


Prognostic signature analysis and survival prediction of esophageal cancer based on N6-methyladenosine associated lncRNAs.
Esophageal cancer (ESCA) has a bad prognosis. Long non-coding RNA (lncRNA) impacts on cell proliferation. However, the prognosis function of N6-methyladenosine (m6A)-associated lncRNAs (m6A-lncRNAs) in ESCA remains unknown. Univariate Cox analysis was applied to investigate prognosis related m6A-lncRNAs, based on which the samples were clustered. Wilcoxon rank and Chi-square tests were adopted to compare the clinical traits, survival, pathway activity and immune infiltration in different clusters where overall survival, clinical traits (N stage), tumor-invasive immune cells and pathway activity were found significantly different. Through least absolute shrinkage and selection operator and proportional hazard (Lasso-Cox) model, five m6A-lncRNAs were selected to construct the prognostic signature (m6A-lncSig) and risk score. To investigate the link between risk score and clinical traits or immunological microenvironments, Chi-square test and Spearman correlation analysis were utilized. Risk score was found connected with N stage, tumor stage, different clusters, macrophages M2, B cells naive and T cells CD4 memory resting. Risk score and tumor stage were found as independent prognostic variables. And the constructed nomogram model had high accuracy in predicting prognosis. The obtained m6A-lncSig could be taken as potential prognostic biomarker for ESCA patients. This study offers a theoretical foundation for clinical diagnosis and prognosis of ESCA.


Developments with experimental and investigational drugs for axial spondyloarthritis.
Axial spondyloarthritis (AxS pA) is a chronic inflammatory disease for which, until recently, there were no valid therapeutic alternatives to TNF-α blocking agents. This unmet clinical need led to explore several therapeutic targets, from proinflammatory cytokines to intracellular signaling systems. The recent approval of Secukinumab, an anti-IL-17A monoclonal antibody, marked a new step in the evolution of AxSpA treatment. Areas covered: the authors review and discuss all the biological or synthetic agents that are currently developed or that have been tested in AxSpA. News from relevant press releases by manufacturers on past, current and future developments are also reported. Several agents that target IL-17 are currently between phase 2 and 3 of clinical development. Ustekinumab, a monoclonal antibody that blocks IL-23 and IL-12 is also in phase 3 after encouraging results from a pilot study. Expert opinion: The advent of agents that target the IL-23/IL-17 axis promises to reshape the therapeutic landscape for AxSpA in the next few years. Open questions in the research agenda for these agents involve their positioning in the therapeutic strategy, their efficacy on the spectrum of skeletal and extraskeletal manifestations of AxSpA, their effect on new bone formation and their long-term tolerance.


Ultrastructure of quick-frozen and freeze-substituted chick osteoclasts.
For comparison with chemically fixed osteoclasts, we prepared chick osteoclasts by quick freezing followed by freeze-substitution. In spite of technical difficulties this demonstrated that osteoclasts can be satisfactorily frozen in situ by the metal contact method. Ultrastructural differences were revealed between conventional fixation and quick freezing. Compared with conventional fixation, the quick freezing method appeared to improve preservation: (1) a discrete trilaminar plasma membrane and other intracellular membranes showed a smooth profile without undulation or rupture; (2) cytoskeletal components appeared to be clearer, straighter, and more numerous; (3) the interior of the ruffled finger contained interconnected lattice structures whereas highly organised microfilaments were seen in the clear zone; (4) well developed tubulovesicular structures (TVSs) that branched or anastomosed with each other were revealed in the cytoplasm; (5) the contents of intracellular membrane systems including the nuclear envelope, endoplasmic reticulum, and Golgi complex were stained to a various extent; (6) vesicles and vacuoles were much smaller, round and well-defined with electron-dense contents; (7) crystalline structures were seen at the extracellular channels of the ruffled border, in the lumen of TVSs, and in vesicles; (8) in some instances mitochondrial granules were visible; (9) within the resorptive lacuna, osteoclasts adhered to the degraded bone matrix without any intervening empty space.


In most poorly controlled glyburide-treated type 2 diabetic patients drug withdrawal causes further increase in glycemia not accompanied by changes in insulin secretion.
To find out whether the secondary failure of glyburide in type 2 diabetes is complete or partial, we studied 38 patients, age (M +/- SD) 69 +/- 9 years, suffering from diabetes from 13.5 +/- 8.4 years and treated with glyburide for 5-13 years, with poor glycemic control (glycohemoglobin 10.6 +/- 2.6%). Serum glucose, insulin and C-peptide were assayed before and 1 h and 2 h after a simulated meal load (355 Cal), after which the drug was replaced with placebo for 4 weeks, and the test repeated. After glyburide withdrawal, fasting glycemia increased from 10.3 +/- 3.3 to 15.1 +/- 4.4 mmol/L (p < 0.001), but in 3/38 patients, it even decreased and in five others the changes were less than +/- 2 mmol/L. These changes negatively but only weakly correlated with initial glycemia: r = 0.4123, p < 0.010. The mean post-meal glycemia at 1 h and 2 h increased respectively by 3.3 and 5.9 mmol/L (both p < 0.001). Neither the levels of glycemia nor its changes after the glyburide withdrawal correlated with the levels of, or changes in, insulin or C-peptide. We conclude that in most but not all type-2 diabetic patients, poorly controlled with glyburide, the drug still retains some limited therapeutic effectiveness, and therefore its withdrawal causes further deterioration of control with the almost equal increases in fasting and post-meal levels of glycemia. These changes are not accompanied by decrease in insulin secretion.


Finding our Place: Making the Connection Toward Faith Integration.
At the onset, the unfamiliarity of faith integration for nurse educators working within a faith-based university can be challenging. Two nurse educators describe the process they took to learn and navigate faith integration, while teaching undergraduate nursing students. Over the course of one year, various approaches toward faith integration with students were implemented, leading to an authentic relational interconnectedness.


Genetic diversity and drug resistance surveillance of Plasmodium falciparum for malaria elimination: is there an ideal tool for resource-limited sub-Saharan Africa?
The intensification of malaria control interventions has resulted in its global decline, but it remains a significant public health burden especially in sub-Saharan Africa (sSA). Knowledge on the parasite diversity, its transmission dynamics, mechanisms of adaptation to environmental and interventional pressures could help refine or develop new control and elimination strategies. Critical to this is the accurate assessment of the parasite's genetic diversity and monitoring of genetic markers of anti-malarial resistance across all susceptible populations. Such wide molecular surveillance will require selected tools and approaches from a variety of ever evolving advancements in technology and the changing epidemiology of malaria. The choice of an effective approach for specific endemic settings remains challenging, particularly for countries in sSA with limited access to advanced technologies. This article examines the current strategies and tools for Plasmodium falciparum genetic diversity typing and resistance monitoring and proposes how the different tools could be employed in resource-poor settings. Advanced approaches enabling targeted deep sequencing is valued as a sensitive method for assessing drug resistance and parasite diversity but remains out of the reach of most laboratories in sSA due to the high cost of development and maintenance. It is, however, feasible to equip a limited number of laboratories as Centres of Excellence in Africa (CEA), which will receive and process samples from a network of peripheral laboratories in the continent. Cheaper, sensitive and portable real-time PCR methods can be used in peripheral laboratories to pre-screen and select samples for targeted deep sequence or genome wide analyses at these CEAs.


Molecular diagnosis of populational variants of Anthonomus grandis (Coleoptera: Curculionidae) in North America.
The utility of the cytochrome oxidase I (COI) DNA sequence used for DNA barcoding and a Sequence Characterized Amplified Region for diagnosing boll weevil, Anthonomus grandis Boheman, variants was evaluated. Maximum likelihood analysis of COI DNA sequences from 154 weevils collected from the United States and Mexico supports previous evidence for limited gene flow between weevil populations on wild cotton and commercial cotton in northern Mexico and southern United States. The wild cotton populations represent a variant of the species called the thurberia weevil, which is not regarded as a significant pest. The 31 boll weevil COI haplotypes observed in the study form two distinct haplogroups (A and B) that are supported by five fixed nucleotide differences and a phylogenetic analysis. Although wild and commercial cotton populations are closely associated with specific haplogroups, there is not a fixed difference between the thurberia weevil variant and other populations. The Sequence Characterized Amplified Region marker generated a larger number of inconclusive results than the COI gene but also supported evidence of shared genotypes between wild and commercial cotton weevil populations. These methods provide additional markers that can assist in the identification of pest weevil populations but not definitively diagnose samples.


Validation of a Method for the Determination of Balenine/Ophidine in Whale.
The aim of this study was to develop and validate a method for the determination of balenine/ophidine (hereafter referred to as "balenine") in whale extracts and muscle samples from Balaenoptera acutorostrata. Further, the goal was to evaluate the method's applicability for the determination of other histidine-containing dipeptides (HCDs): anserine and carnosine and their amino acids π-methylhistidine, τ-methylhistidine, histidine, and β-alanine. For balenine, the LOD and LOQ were found to be 0.03 and 0.1 mg/g, respectively, and the linear range was validated up to 160 mg/g. Trueness was evaluated by spiking experiments with balenine, and the recovery was found to be 88-90%. A comparison of the results showed that most of the other analytes were within 80-120% of the value found with the previously developed and validated method. Precision and internal reproducibility for balenine was around 0.9 and 2%, respectively, with measurement uncertainties of 2-4%. Therefore, the method was found to be fit for purpose for the determination of balenine and other HCDs and their constituent amino acids in whale meat and extracts.


One-step measurement of firefly luciferase activity in yeast.
Firefly luciferase is often used as a sensitive genetic reporter in various cell types. The pitfall in yeast, however, has been the need to break down the rigid cells in order to measure the enzyme activity. In this study we have removed the peroxisomal targeting codons from the Photinus pyralis luciferase gene (luc) and shown that in the yeast Saccharomyces cerevisiae this modified luciferase gives high levels of light emission that is easy to measure from intact living cells. Furthermore, cells with the modified luciferase grew essentially faster than those with the wild-type luciferase, indicating that peroxisomal targeting of a foreign enzyme puts some constraints to cellular viability. As a model system we used two different reporter constructs. In the first, expression of the luciferase gene is under control of CUP1-promoter, a well known yeast promoter that is inducible by copper ions. In the second, luciferase activity is dependent on activation of the human oestrogen receptor and its interaction with oestrogen-responsive elements incorporated in a yeast promoter. The luciferase activity measurement could be done on a 96-well plate by simple addition of the substrate, D-luciferin, at a moderately acidic pH of 5.0. The ease of use of the non-peroxisomal luciferase makes it an interesting alternative for reporter genes that are conventionally used in yeast, such as lacZ.


Polymorphism and synergism of angiotensin-converting enzyme (ACE) and plasminogen activator inhibitor-1 (PAI-1) genes in coronary artery disease.
Among the genetic factors for coronary artery diseases, PAI-1 4G/5G and ACE I/D polymorphisms can be noted. This study was carried out to investigate the association of these two polymorphisms and their synergism in coronary artery disease (CAD) from a sample of the Iranian population. Sixty-one patients with a history of CAD and 92 healthy controls participated in our study. After DNA extraction from leukocytes, PCR was performed to characterize PAI-1 4G/5G and ACE I/D polymorphisms, using an amplification refractory mutation system technique. In the studied patients, PAI-1 polymorphisms were 24.6%, 45.9%, and 29.5% for 4G/4G, 4G/5G and 5G/5G, respectively; the values for controls were 20.7%, 42.2% and 37.0%. The distribution rates of genotypes I/I, I/D and D/D in patients accounted for 29.5%, 45.9% and 24.6%; in the control group these figures were estimated to be 40.2%, 40.2% and 19.6%. Single and multivariate analyses showed a significant difference for the conventional risk factors, including hypertension, diabetes, hyperlipidemia, smoking and family history, for CAD between patients and controls (p value ≤ 0.001). However, no significant correlation was demonstrated considering ACE and PAI-1 polymorphisms either in association with 4G/4G or D/D genotypes or a combination of them in the Iranian population in the current study.


Cone calix[4]arene diethyl ester strapped calix[4]pyrrole: a selective receptor for the fluoride anion.
A calix[4]pyrrole strapped by a bulky calix[4]arene diester locked in its cone conformation has been prepared. On the basis of (1)H NMR spectroscopic analyses carried out in CDCl3, it is concluded that this hybrid system, receptor 2, binds only the fluoride anion and does so with remarkably high affinity even in the presence of an excess of various potentially competing environmentally and biologically ubiquitous anions (studied as the tetrabutylammonium, cesium, or lithium salts). Solid state structural analyses provide support for the notion that receptor 2 interacts well with the fluoride anion in the solid state.


The Potential for Topical Probiotic Treatment of Chronic Rhinosinusitis, a Personal Perspective.
This review describes the rationale for topical probiotic intervention, the obstacles we are facing and a strategy for future research in the use of probiotics to modify CRS symptoms and disease expression. Recent advances in molecular microbiology has revealed a plethora of microbial DNA in the nasal cavity and sinuses of healthy subjects as well as in chronic sinusitis (CRS) patients. An infection is today rather seen as an imbalance between the commensal microbiome and the bacterial pathogens, resulting in a reduction in commensal bacterial diversity, combined with an increase in the growth of microbes eliciting an inflammatory response. This will in turn lead to the clinical symptoms of sinusitis. Probiotics (microorganisms that confer a health benefit) can be used either as a form of living antibiotics treatment, or as an immune-modulatory intervention. Topical probiotics, which is the focus of this review, have shown efficacy in a limited number of trials in otitis media and tonsillitis, but to date not in CRS. Although bacterial interference capacity against pathogens can be determined in in vitro experiments, it may not translate to a health benefit. This limits the role of laboratory research in identifying probiotic strains with a clinical benefit. To gain more clinical experience without further delay, I recommend future research to focus on empirical clinical trials in well-defined CRS patient populations and study the underlying mechanisms in more detail once a clinical benefit has been established.


[Relationship of nitric oxide with the apoptosis of keratinocytes].
Nitric oxide (NO) is a kind of free radical, also is a signaling molecule, which universally distributed in the tissues of organism. NO play important role in the dermato-, cardia-, encephalo-, hepat- and immunological regulation on the physio- and patho- situation. Recently studying show that NO can bifunctionally regulate the apoptosis of keratinocytes, which play an important role in the occurrence and development of many skin diseases. However, the regulatory mechanism still can't be completely elucidated at present time. In this review, we are reviewing the recent investigation progress on the apoptosis of keratinocytes and some diseases of skin that related to the regulating disturbance of NO.


Isolation and characterization of human urinary kallikrein.
Human urinary kallikrein was purified by gel filtration on Sephacryl S-200 and affinity chromatography on aprotinin-Sepharose, followed by ion exchange chromatography on DEAE-Sepharose. Thus an enzyme preparation with a specific activity (using AcPheArgOEt as substrate) of 1 100 U/mg protein was obtained. A specific bioligical activity of 2 300 KE/mg was measured in the dog blood pressure assay and of 0.742 HMW kininogen-U/mg, corresponding to the liberation of 787 micrograms bradykinin per mg enzyme per min from HMW-kininogen, in the rat uterus assay. In dodecyl sulfate electrophoresis two protein bands with apparent molecular weights of 41 000 and 34 000 were separated. The amino acid composition was determined and isoleucine was identified as the only aminoterminal amino acid. On isoelectric focusing six protein bands with isoelectric points of 3.75, 3.80, 3.90, 4.00, 4.05 and 4.25 were separated. The kinetic constants for the kallikrein-catalyzed hyrdolysis of AcPheArgOEt and D ValLeuArgNHNp were determined. The bimolecular velocity constant for the inhibition by diisopropyl fluorophosphate was determined as 9 +/- 2l x mol-1 x min-1. Immunological studies showed that a close relationship exists between the urinary enzyme and other human glandular kallikreins. Deoxycholate, lysolecithin and other amphiphiles activated human urinary kallikrein.


Circulating copeptin level and the clinical prognosis of patients with chronic liver disease.
The relationship between copeptin and the severity of circulatory dysfunction and systemic stress response in patients with chronic liver disease (CLD) has been established. Nevertheless, the potential of serum copeptin levels to predict the prognosis of CLD patients remains unclear. To conduct a systematic review and meta-analysis to investigate the correlation between serum copeptin and transplant-free survival (TFS) in this population. To achieve the objective of the meta-analysis, PubMed, Embase, the Cochrane Library, and the Web of Science were searched to identify observational studies with longitudinal follow-up. The Cochrane Q test was utilized to assess between-study heterogeneity, and the I2 statistic was estimated. Random-effects models were employed to combine the outcomes, taking into account the potential influence of heterogeneity. Ten datasets including 3133 patients were involved. The follow-up durations were 1 to 48 mo (mean: 12.5 mo). Overall, it was shown that a high level of serum copeptin was associated with a poor TFS [risk ratio (RR): 1.82, 95% confidence interval: 1.52-2.19, P < 0.001; I2 = 0%]. In addition, sensitivity analysis by omitting one dataset at a time showed consistent results (RR: 1.73-2.00, P < 0.05). Finally, subgroup analyses according to study country, study design, patient diagnosis, cutoff of copeptin, follow-up duration, and study quality score also showed similar results (P for subgroup difference all > 0.05). Patients with CLD who have high serum copeptin concentrations may be associated with a poor clinical prognosis.


Effect of in-feed inclusion of a natural zeolite (clinoptilolite) on certain vitamin, macro and trace element concentrations in the blood, liver and kidney tissues of sows.
The study was conducted to evaluate, under field conditions, the effect of the long-term dietary use of a natural zeolite (clinoptilolite, CLI) and antibacterials (chlortetracycline, CTC) on the concentrations of certain vitamins (vitamin A and vitamin E) and minerals (K, Na, P, Ca, Mg, Cu and Zn) in blood and body tissues of the sow. Twenty-four sows were assigned to two main experimental groups and four subgroups, depending on the presence or absence of CLI and CTC in their feed, respectively. CLI was provided to the sows from weaning, during the service, gestation and lactation periods and up to the date of the next service, while CTC was administered for a 2-week period post-service, as well as for a 2-week period following the allocation of the sows in the farrowing house, around 5 days prior to the expected parturition. Blood samples were collected on the starting day of the trial, on the 30th and the 90th day of each pregnancy, on the day of each parturition and on the day of each weaning. Furthermore, 20 sows were similarly distributed in the same experimental groups and subgroups and at the end of the trial they were slaughtered and liver and kidney samples were collected for biochemical analysis. Neither CLI nor CTC supplementation of the diets had any significant effect on vitamins' and minerals' uptake and their distribution in the body, since there was no alteration in their blood serum and liver/kidney concentrations. Furthermore, no CLI x CTC interaction was noticed.


Identification of cell types responsible for bone resorption in rheumatoid arthritis and juvenile rheumatoid arthritis.
Focal resorption of bone at the bone-pannus interface is common in rheumatoid arthritis (RA) and juvenile rheumatoid arthritis (JRA) and can result in significant morbidity. However, the specific cellular and hormonal mechanisms involved in this process are not well established. We examined tissue sections from areas of bone erosion in patients with RA and JRA. Multinucleated cells (MNCs) were present in resorption lacunae in areas of calcified cartilage and in subchondral bone immediately adjacent to calcified cartilage, as previously described. mRNA for the calcitonin receptor (CTR) was localized to these MNCs in bone resorption lacunae, a finding that definitively identifies these cells as osteoclasts. These MNCs were also positive for tartrate-resistant acid phosphatase (TRAP) mRNA and TRAP enzymatic activity. Occasional mononuclear cells on the bone surface were also CTR positive. Mononuclear cells and MNCs not on bone surfaces were CTR negative. The restriction of CTR-positive cells to the surface of mineralized tissues suggests that bone and/or calcified cartilage provide signals that are critical for the differentiation of hematopoietic osteoclast precursors to fully differentiated osteoclasts. Some MNCs and mononuclear cells off bone and within invading tissues were TRAP positive. These cells likely represent the precursors of the CTR-TRAP-positive cells on bone. Parathyroid hormone receptor mRNA was present in cells with the phenotypic appearance of osteoblasts, in close proximity to MNCs, and in occasional cells within pannus tissue, but not in the MNCs in bone resorption lacunae. These findings demonstrate that osteoclasts within the rheumatoid lesion do not express parathyroid hormone receptor. In conclusion, the resorbing cells in RA exhibit a definitive osteoclastic phenotype, suggesting that pharmacological agents that inhibit osteoclast recruitment or activity are rational targets for blocking focal bone erosion in patients with RA and JRA.


The genesis and implementation of the Tasmanian obstetric survey.
The genesis and implementation of a continuing obstetric audit in Tasmania is described. The survey includes all public and private deliveries in Tasmania. Some of the early difficulties and their resolution are discussed. Because of its simplicity, the suggestion has been made that such a system would be suitable for a national audit.


Remote Generation of Wigner Negativity through Einstein-Podolsky-Rosen Steering.
Negativity of the Wigner function is seen as a crucial resource for reaching a quantum computational advantage with continuous variable systems. However, these systems, while they allow for the deterministic generation of large entangled states, require an extra element such as photon subtraction to obtain such negativity. Photon subtraction is known to affect modes beyond the one where the photon is subtracted, an effect which is governed by the correlations of the state. In this Letter, we build upon this effect to remotely prepare states with Wigner negativity. More specifically, we show that photon subtraction can induce Wigner negativity in a correlated mode if and only if that correlated mode can perform Einstein-Podolsky-Rosen steering in the mode of subtraction.


Regional circadian period difference in the suprachiasmatic nucleus of the mammalian circadian center.
The suprachiasmatic nucleus (SCN) is the mammalian circadian rhythm center. Individual oscillating neurons have different endogenous circadian periods, but they are usually synchronized by an intercellular coupling mechanism. The differences in the period of each oscillating neuron have been extensively studied; however, the clustering of oscillators with similar periods has not been reported. In the present study, we artificially disrupted the intercellular coupling among oscillating neurons in the SCN and observed regional differences in the periods of the oscillating small-latticed regions of the SCN using a transgenic rat carrying a luciferase reporter gene driven by regulatory elements from a per2 clock gene (Per2::dluc rat). The analysis divided the SCN into two regions--aregion with periods shorter than 24 h (short-period region, SPR) and another with periods longer than 24 h (long-period region, LPR). The SPR was located in the smaller medial region of the dorsal SCN, whereas the LPR occupied the remaining larger region. We also found that slices containing the medial region of the SCN generated shorter circadian periods than slices that contained the lateral region of the SCN. Interestingly, the SPR corresponded well with the region where the SCN phase wave is generated. We numerically simulated the relationship between the SPR and a large LPR. A mathematical model of the SCN based on our findings faithfully reproduced the kinetics of the oscillators in the SCN in synchronized conditions, assuming the existence of clustered short-period oscillators.


The effects of extinction-aroused attention on context conditioning.
Two experiments assessed the effects of extinguishing a conditioned cue on subsequent context conditioning. Each experiment used a different video-game method where sensors predicted attacking spaceships and participants responded to the sensor in a way that prepared them for the upcoming attack. In Experiment 1 extinction of a cue which signaled a spaceship-attack outcome facilitated subsequent learning when the attack occurred unsignaled. In Experiment 2 extinction of a cue facilitated subsequent learning, regardless of whether the spaceship outcome was the same or different as used in the earlier training. In neither experiment did the extinction context become inhibitory. Results are discussed in terms of current associative theories of attention and conditioning.


Cerebral arterial gas embolism in a diver using closed circuit rebreather diving apparatus.
A Singaporean naval diver suffered symptoms and signs of cerebral arterial gas embolism (CAGE) following a panicked ascent from a depth of 5-6 metres' sea water while using a Dräger LAR V closed-circuit oxygen rebreather system. He presented with altered mental status and paresis. CAGE due to gas mixtures with high oxygen content has seldom been reported. The diver had no sequelae following prompt recompression therapy. This positive clinical outcome may be attributable to the high oxygen content in the diver's inspired gas and/or the promptness of recompression.


Time course of 8-methoxypsoralen-induced skin photosensitization in PUVA-bath photochemotherapy.
In recent years PUVA-bath photochemotherapy has been shown to be an effective treatment modality for several dermatoses. A limitation of PUVA-bath photochemotherapy has been the lack of guidelines for optimal performance, including the time course of photosensitization of the skin exposed to the 8-methoxypsoralen (8-MOP) bath water solution. In the present study 12 healthy volunteers were exposed to a 20 min bath in 150 l of an 8-MOP water solution (0.5 mg/l, 37 degrees C). Immediately, as well as 1, 2, 3 and 5 h after the 8-MOP bath, irradiation was performed with increasing doses of UVA (0.5, 1, 2, 3, 5 J/cm2) on 2 cm2 test areas. The minimal phototoxic dose (MPD) was determined 72 h after the UVA exposure. In all volunteers, photosensitization was highest immediately after the bath, with a MPD significantly below 5 J/cm2 (0.5-2 J/cm2). One hour after the bath, erythema could be induced by 2 to 5 J/cm2 UVA. Two hours after the bath, erythema could be induced using irradiation of 5 J/cm2 only in two volunteers. Three and five hours after the 8-MOP bath, no erythema could be induced in any volunteer by UVA doses up to 5 J/cm2. Our results indicate that optimal bath-PUVA requires UVA irradiation immediately after the 8-MOP bath. Further, these results imply that no restrictions on further sun exposure are mandatory 3 h after the 8-MOP bath, thus allowing the patient to pursue normal life activities.


Influence of primary payer status on non-ST-segment elevation myocardial infarction: 18-year retrospective cohort national temporal trends, management and outcomes.
The role of insurance on outcomes in non-ST-segment-elevation myocardial infarction (NSTEMI) patients is limited in the contemporary era. From the National Inpatient Sample, adult NSTEMI admissions were identified [2000-2017]. Expected primary payer was classified into Medicare, Medicaid, private, uninsured and others. Outcomes included in-hospital mortality, overall and early coronary angiography, percutaneous coronary intervention (PCI), resource utilization and discharge disposition. Of the 7,290,565 NSTEMI admissions, Medicare, Medicaid, private, uninsured and other insurances were noted in 62.9%, 6.1%, 24.1%, 4.6% and 2.3%, respectively. Compared to others, those with Medicare insurance older (76 vs. 53-60 years), more likely to be female (48% vs. 25-44%), of white race, and with higher comorbidity (all P<0.001). Population from the Medicare cohort had higher in-hospital mortality (5.6%) compared to the others (1.9-3.4%), P<0.001. With Medicare as referent, in-hospital mortality was higher in other {adjusted odds ratio (aOR) 1.15 [95% confidence interval (CI), 1.11-1.19]; P<0.001}, and lower in Medicaid [aOR 0.95 (95% CI, 0.92-0.97); P<0.001], private [aOR 0.77 (95% CI, 0.75-0.78); P<0.001] and uninsured cohorts [aOR 0.97 (95% CI, 0.94-1.00); P=0.06] in a multivariable analysis. Coronary angiography (overall 52% vs. 65-74%; early 15% vs. 22-27%) and PCI (27% vs. 35-44%) were used lesser in the Medicare population. The Medicare population had longer lengths of stay, lowest hospitalization costs and fewer home discharges. Compared to other types of primary payers, NSTEMI admissions with Medicare insurance had lower use of coronary angiography and PCI, and higher in-hospital mortality.


Role of the extracellular matrix in epithelial morphogenesis: a view from C. elegans.
The extracellular matrix (ECM) plays an essential role in organizing tissues, defining their shapes or in presenting growth factors. Their components have been well described in most species, but our understanding of the mechanisms that control ECM remodeling remains limited. Likewise, how the ECM contributes to cellular mechanical responses has been examined in few cases. Here, I review how studies performed in C. elegans have brought several significant advances on those topics. Focusing only on epithelial cells, I discuss basement membrane invasion by the anchor cell during vulva morphogenesis, a process that has greatly expanded our knowledge of ECM remodeling in vivo. I then discuss the ECM role in a novel mechanotransduction process, whereby muscle contractions stimulate the remodeling of hemidesmosome-like junctions in the epidermis, which highlights that these junctions are mechanosensitive. Finally, I discuss progress in defining the composition and potential roles of the apical ECM covering epidermal cells in embryos.


Dual Roles of Nicotinamide Phosphoribosyltransferase as a Promising Target for Cancer Radiotherapy.
Nicotinamide phosphoribosyltransferase (NAMPT) is the key rate-limiting enzyme in the regulation of nicotinamide adenine dinucleotide (NAD) biosynthesis, and its activity is critical for the replenishment of NAD level as well as cell survival or death. As one of the most important components in the electron transport chain of complex I in mitochondrion, sustained supply of NAD is essential to the maintenance of energy metabolism both in normal and cancer cells. Recent research showed that X-ray radiation sharply downregulated the expression of NAMPT, which may be the main cause of radiation damage in salivary gland. Consistently, upregulation of NAMPT by phenylephrine restored the function and tissue structure of salivary gland, indicating the cytoprotective role of NAMPT in preventing radiation damage in normal tissues of patients with head and neck cancer during radiotherapy. On the other hand, NAMPT downregulation and NAD depletion could induce cell death in oral squamous cell cancer, suggesting that a combination of NAMPT inhibitor and radiotherapy presents a promising therapeutic strategy for cancer treatment. Based on our and other's studies, NAMPT may have dual roles in cancer radiotherapy: the upregulation of NAMPT could serve to suppress radiotherapy complications such as radiation sialadenitis, and combination regimens that involve NAMPT inhibitors may enhance efficacy of radiotherapy for cancer treatment.


[Permanence of the piamontese culture on food practices in third generation descendants in Piamonte (Province of Santa Fe, Argentina)].
This paper describes the influence of the piemontese culture on food practices of students living in Piamonte, Santa Fe, Argentina. Food practices of 96 students with Piamontese ancestry (PA) (n = 57) and without Piamontese ancestry (No-PA) (n = 39) were studied along 2002 using a self-administered questionnare. Data were analysed by Chi square test, Fisher's exact test, multiple correspondance analysis and logistic regression. Consumption of bagna cauda (p < 0.05) and polenta (p < 0.1) were higher among PA. Differences on the elaboration of polenta and pasta were found: PA add them cheese (p < 0.05) and cream (p < 0.05) whereas no-PA make use of meat (p < 0.05) and tomato sauce (p < 0.05), respectively. The frequency of consumption of traditional Piamontese meals and the role of the mother in the purchase, the elaboration and the serving of the food were similar on both groups. In conclusion, food practices of Piamontese's descendants recall the food culture of their ancestry with some reasonable adaptations to the local context.


Investigation of the Effect of Different Emulsifiers on the Transdermal Delivery of EGCG Entrapped in a Polymeric Micelle System.
Epigallocatechin gallate, one of the most active antioxidant compounds, has a low chemical stability and ability to permeate the human epidermis. The encapsulation in polymeric micelles would be beneficial to improve both stability and permeation of epigallocatechin gallate and, therefore, to facilitate the pharmacological effects. Polymeric micelles containing epigallocatechin gallate were incorporated in O/W emulsions prepared by using different types of emulsifying systems. All emulsions were uniform in colour and aspect, without evidences of phase separation after centrifugation at the preparation time and over a 6-month period of storage at room temperature. Emulsions containing epigallocatechin gallate incorporated in polymeric micelles showed a colour variation, probably due to epigallocatechin gallate degradation, over the stability period. The skin permeability study evidenced a significant increase in epigallocatechin gallate permeation after encapsulation in micelles. Pure epigallocatechin gallate was not able to permeate the skin and only limited amounts were retained in the epidermis, while both permeated and retained amounts after 24 h were measured in the case of polymeric micelles containing epigallocatechin gallate. Moreover, the epigallocatechin gallate release and human skin permeability were affected by the type of emulsifier. The epigallocatechin gallate release in the presence of an emulsifier system based on cereal and fruit fibres never occurred. The best results in terms of release and skin permeability were obtained using glycerides of synthetic or semisynthetic origin or esters.


Development and Transferability of EST-SSR Markers for Pinus koraiensis from Cold-Stressed Transcriptome through Illumina Sequencing.
Pinus koraiensis has significant economic and ecological value in Northeast China. However, due to the lack of suitable molecular markers, only a few available microsatellite markers were developed for further population genetics studies. In this study, for the first time we developed expressed sequence tag-simple sequence repeat (EST-SSR) markers from the cold-stressed transcriptome of P. koraiensis using Illumina Sequencing. We identified a total of 7,235 EST-SSRs from 97,376 sequences, and we tested their transferability among seven related Pinus species. The results showed that trinucleotides were the most abundant type of repeat (1287, 18.74%) excluding mononucleotides, followed by dinucleotides (1284, 18.7%) and tetranucleotides (72, 1.05%). The most dominant dinucleotides and trinucleotide repeat motifs were AT/AT (535, 7.79%) and AAT/ATT (103, 1.5%). The observed heterozygosity (Ho) and expected heterozygosity (He) ranged from 0.002 to 0.986 and 0.017 to 0.743, respectively, and the polymorphism information content (PIC) values and number of alleles (Na) varied from 0.029 to 0.794 and 2 to 23, respectively. A total of 8 natural P. koraiensis populations were divided into two main genetic clusters. Furthermore, nine of twenty polymorphic primer pairs were successfully amplified in seven Pinus species, and at least 80% of the successful P. koraiensis EST-SSR primers could be amplified in more than four species (16, 80%). Combined results for the development of EST-SSR markers in P. koraiensis and transferability among related species would contribute to improved studies on the genetic diversity and population structure in P. koraiensis and phylogenetic relationships among Pinus species. They would also provide a significant source for quantitative trait locus analysis.


Retrospective Molecular Epidemiology Study of PD-L1 Expression in Patients with EGFR-Mutant Non-small Cell Lung Cancer.
Data are limited on programmed death ligand 1 (PD-L1) expression in epidermal growth factor receptor (EGFR)-mutant non-small cell lung cancer (NSCLC). We retrospectively evaluated the relationship between PD-L1 expression and recurrence-free survival (RFS) and overall survival in 319 patients with EGFR-mutant NSCLC who were treated at Samsung Medical Center from 2006 to 2014. Membranous PD-L1 expression on tumor cells was measured using the PD-L1 IHC 22C3 pharmDx antibody and reported as tumor proportion score (TPS). Kaplan-Meier methods, log-rank test, and Cox proportional hazards models were used for survival analysis. All patients had ≥1 EGFR mutation-54% in exon 19 and 39% in exon 21. Overall, 51% of patients had PD-L1-positive tumors. The prevalence of PD-L1 positivity was higher among patients with stages II-IV versus stage I disease (64% vs. 44%) and among patients with other EGFR mutations (75%) than with L858R mutation (39%) or exon 19 deletion (52%). PD-L1 positivity was associated with shorter RFS, with an adjusted hazard ratio of 1.52 (95% confidence interval [CI], 0.81 to 2.84; median, 18 months) for the PD-L1 TPS ≥ 50% group, 1.51 (95% CI, 1.02 to 2.21; median, 31 months) for the PD-L1 TPS 1%-49% group, and 1.51 (95% CI, 1.05 to 2.18) for the combined PD-L1-positive groups (TPS ≥ 1%) compared with the PD-L1-negative group (median, 35 months). PD-L1 expression is associated with disease stage and type of EGFR mutation. PD-L1 positivity might be associated with worse RFS among patients with surgically treated EGFR-mutant NSCLC.


Synthesis and biological evaluation of quinoline salicylic acids as P-selectin antagonists.
Leukocyte recruitment of sites of inflammation and tissue injury involves leukocyte rolling along the endothelial wall, followed by firm adherence of the leukocyte, and finally transmigration of the leukocyte across cell junctions into the underlying tissue. The initial rolling step is mediated by the interaction of leukocyte glycoproteins containing active moieties such as sialyl Lewisx (sLex) with P-selectin expressed on endothelial cells. Consequently, inhibition of this interaction by means of a small molecule P-selectin antagonist is an attractive strategy for the treatment of inflammatory diseases such as arthritis. High-throughput screening of the Wyeth chemical library identified the quinoline salicylic acid class of compounds (1) as antagonists of P-selectin, with potency in in vitro and cell-based assays far superior to that of sLex. Through iterative medicinal chemistry, we identified analogues with improved P-selectin activity, decreased inhibition of dihydrooratate dehydrogenase, and acceptable CYP profiles. Lead compound 36 was efficacious in the rat AIA model of rheumatoid arthritis.


Expression of Ki-67, Cyclin D1, P53, and P16 in patients with oral leukoplakia and leukoplakia cancerization with spicy diet in Chengdu.
To investigate the expression of Ki-67, Cyclin D1, P53, and P16 in patients with oral leukoplakia (OLK) and OLK cancerization who have aspicy diet in Chengdu. Thirtypatients with OLK andspicy diet and 15 patients with OLK without spicy diet in Chengdu were divided into three groups: hyperplastic OLK (OLK-), OLK with mild to moderate dysplasia (OLK+), and severe dysplastic OLK or oral squamous cell carcinoma (OSCC) transforming from OLK (OLK++/OSCC). The expression of Ki-67, Cyclin D1, P53, and P16 were detected by immunohistochemistry and statistically analyzed. The expression of Ki-67 and P53 in patients with or without spicy diet in the OLK+and OLK++/OSCC groups were stronger than that of the OLK- group (P<0.05). The OLK++/OSCC group showed a higher expression of Cyclin D1 and lower expression of P16 than the OLK- group (P<0.05). The expression of Ki-67, Cyclin D1, P53, and P16 in patients with spicy diet and without spicy diet had no substantial difference. The expression of Ki-67 and Cyclin D1 showed a positive correlation (r=0.439, P=0.015). Spicy diet did not have an influence on the expression of Ki-67, Cyclin D1, P53, and P16 in patients with OLK and OSCC. The expression of Ki-67, Cyclin D1, and P53 increased with the development of OLK, whereas P16 showed opposite expression trend.


Whole-Exome Sequencing Identifies Loci Associated with Blood Cell Traits and Reveals a Role for Alternative GFI1B Splice Variants in Human Hematopoiesis.
Circulating blood cell counts and indices are important indicators of hematopoietic function and a number of clinical parameters, such as blood oxygen-carrying capacity, inflammation, and hemostasis. By performing whole-exome sequence association analyses of hematologic quantitative traits in 15,459 community-dwelling individuals, followed by in silico replication in up to 52,024 independent samples, we identified two previously undescribed coding variants associated with lower platelet count: a common missense variant in CPS1 (rs1047891, MAF = 0.33, discovery + replication p = 6.38 × 10(-10)) and a rare synonymous variant in GFI1B (rs150813342, MAF = 0.009, discovery + replication p = 1.79 × 10(-27)). By performing CRISPR/Cas9 genome editing in hematopoietic cell lines and follow-up targeted knockdown experiments in primary human hematopoietic stem and progenitor cells, we demonstrate an alternative splicing mechanism by which the GFI1B rs150813342 variant suppresses formation of a GFI1B isoform that preferentially promotes megakaryocyte differentiation and platelet production. These results demonstrate how unbiased studies of natural variation in blood cell traits can provide insight into the regulation of human hematopoiesis.


[Prognostic significance of doppler-derived and left ventricular diastolic filling variables in patients with dilated cardiomyopathy].
The purpose of this echocardiographic study was to evaluate the frequency of restrictive left ventricular filling pattern in dilated cardiomyopathy, as well as its clinical and echocardiographic correlations and prognostic implications. The study included 49 pts (39 males, 10 females) aged 32 to 56 years (mean age: 46 years), with a diagnosis of dilated cardiomyopathy, according to the World Health Organization criteria. Pulsed Doppler echocardiographic analysis was performed at discharge and during a two year follow-up and compared with clinical and 2-dimensional echocardiographic findings. The patients were divided into two groups according to measurement of E decelaration time (DT) of transmitral flow patterns: a restrictive group (Group A-21 pts) with DT < 115 ms. and a non-restrictive group (Group B-28 pts) with DT > 115 ms. Of 49 pts, 15 died during a two-year follow-up, 12 in the restrictive group and only 3 in the non-restrictive group. Eleven of these pts (73%) died of worsening of congestive heart failure and four (27%) died suddenly. The restrictive filling pattern was associated at Doppler study with a higher E wave velocity, lower A wave velocity and higher E/A ratio. Pts in the restrictive group were in a higher New York Heart Association functional class, and had a lower ejection fraction and more severe mitral regurgitation. In addition, they had a significantly larger right ventricle and higher mean pulmonary artery pressure. Mortality rate in the restrictive group was markedly higher than that in the non-restrictive group at 1 year (24% vs. 0%, respectively, p < 0.001) and at 2 years (57% vs. 11%, respectively, p < 0.001). The results from this study indicate that a restrictive left ventricular filling pattern is frequent in dilated cardiomyopathy, and is associated with more severe disease and is the single best clinical predictor for cardiac death in pts with dilated cardiomyopathy.


Preface to special topic: dielectrophoresis.
This Special Topic section is on dielectrophoresis, a growing area of widespread interest and relevance to the microfluidics and nanofluidics community.


[Preproghrelin gene, ghrelin receptor and metabolic syndrome].
Obesity is a multi-gene syndrome, expression of which is modulated not only by environmental factors but above all by a number of modified genes interacting with each other. Among candidate genes related to obesity phenotype is ghrelin gene. Ghrelin plays a significant role in feeding regulation and is the strongest stimulator of growth hormone secretion. Ghrelin acts by GHS1a receptor (growth hormone secretagogue receptor). Mutations in preproghrelin and ghrelin gene or ghrelin receptor gene could be responsible for low ghrelin levels observed in obese individuals. Among identificated mutations, two Arg51 Gln and Leu72Met are most often described and change amino-acid sequence of ghrelin (Arg51Gln) and preproghrelin (Leu72Met). Although no direct relationship between Arg51Gln mutation and obesity phenotype was found, it had been shown that carriers of Arg51Gln mutation had significantly decreased plasma ghrelin levels. Furthermore 51Gln allele carriers had higher prevalence of type 2 diabetes mellitus and hypertension than non-carriers. Met 72 carrier status is associated with higher serum IGF-1 levels and seems to be a protective factor against fat accumulation and cardiovascular complications of obesity. No evidence of relationship between ghrelin receptor gene polymorphisms and body mass regulation was found, however, until now there is no study on relationships between these polymorphisms and metabolic complications of obesity. The presence of genetic variants in ghrelin or GHS receptor gene could be responsible for impaired GH secretion in visceral type obesity and development of metabolic syndrome in some of obese subjects. On the other hand, some mutations in preproghrelin gene could be protective against metabolic syndrome.


Ceftriaxone versus latamoxef in febrile neutropenic patients: empirical monotherapy in patients with solid tumours.
121 patients with 132 febrile episodes were randomised to ceftriaxone or latamoxef monotherapy in order to compare antibiotic efficacy in neutropenic patients treated with cytotoxic chemotherapy for solid tumours. In 80 evaluable episodes no significant differences were observed between the two groups with respect to efficacy and fatal failure rates. Of episodes treated with ceftriaxone, 67% showed a favourable clinical response vs. 61% in the latamoxef group. The clinical response rates in episodes with documented bacterial infections were 67 and 56% in the two treatment groups. In 18% of the episodes with documented initial infections the patients died of presumably uncontrolled infection. The convenient once daily dosage schedule combined with fewer severe adverse reactions favours the use of ceftriaxone instead of latamoxef. Although a relative high degree of response was seen, empirical antibiotic monotherapy apparently does not offer a sufficient antibacterial cover in infections in this type of patient with defective host immunity.


OpenDEP: An Open-Source Platform for Dielectrophoresis Spectra Acquisition and Analysis.
Dielectrophoretic (DEP) cell separation, which utilizes electric fields to selectively manipulate and separate cells based on their electrical properties, has emerged as a cutting-edge label-free technique. DEP separation techniques rely on differences in the electrical and morphological properties of cells, which can be obtained by a thorough analysis of DEP spectra. This article presents a novel platform, named OpenDEP, for acquiring and processing DEP spectra of suspended cells. The platform consists of lab-on-a-chip and open-source software that enables the determination of DEP spectra and electric parameters. The performance of OpenDEP was validated by comparing the results obtained using this platform with the results obtained using a commercially available device, 3DEP from DEPtech. The lab-on-a-chip design features two indium tin oxide-coated slides with a specific geometry, forming a chamber where cells are exposed to an inhomogeneous alternating electric field with different frequencies, and microscopic images of cell distributions are acquired. A custom-built software written in the Python programing language was developed to convert the acquired images into DEP spectra, allowing for the estimation of membrane and cytoplasm conductivities and permittivities. The platform was validated using two cell lines, DC3F and NIH 3T3. The OpenDEP platform offers several advantages, including easy manufacturing, statistically robust computations due to large cell population analysis, and a closed environment for sterile work. Furthermore, continuous observation using any microscope allows for integration with other techniques.


ZC4H2 deletions can cause severe phenotype in female carriers.
ZC4H2 is involved in human brain development, and, if mutated, can be responsible for a rare X-linked disorder, originally presented in literature as Wieacker-Wolff syndrome and Miles-Carpenter syndrome. In males, severe intellectual disability is associated with variable symptoms of central and peripheral nervous system involvement, such as spasticity, hyperreflexia, muscle weakness, and arthrogryposis. Female carriers are usually described as asymptomatic or only mildly affected. Here, we report on a girl carrying a de novo deletion of ZC4H2 detected by array-CGH analysis. She showed a complex neurodevelopmental disorder resembling the clinical picture commonly observed in male patients. X-inactivation was found to be random. Additionally, she had an unusual appearance of fingers and hand creases, and electromyography showed a peculiar pattern of both neurogenic and myopathic anomalies. The present patient confirms that female carriers can also be severely affected. Systematic clinical investigations of both males and females are needed to define the variety in nature and severity of phenotypes related to ZC4H2 variants.


The "inherent vice" in the anti-angiogenic theory may cause the highly metastatic cancer to spread more aggressively.
Although anti-angiogenic (AA) therapy is widely used in clinical practice, it is often challenged by insufficient efficacy and intrinsic resistance. Some studies have reported that AA therapy can even increase tumor metastasis. However, whether this is due to a specific AA drug causing a specific tumor to metastasize or because the anti-angiogenic theory has some "inherent vice" that may inevitably lead to tumor dissemination remains a mystery. Herein, we designed a model that completely blocks tumor blood supply using a physical barrier to examine tumor behavior in such circumstances. Surprisingly, we found that cutting off the blood supply could neither eliminate the primary tumor cells nor prevent local invasion or formation of distant metastases. By using a mathematical method to simulate tumor behavior, we found that blocking tumor blood supply may lead to an inevitable consequence: the cells that can tolerate blood deficiency are "naturally selected" and survive, whereas a portion of cells are promoted to escape from the "starvation" area by the consistent environmental stress until they are spread throughout the body. This may be an intrinsic disadvantage of the AA strategy, which will inevitably cause the tumor, particularly highly metastatic tumors, to spread more aggressively.


Rapid mixing with high-throughput in a semi-active semi-passive micromixer.
In this paper, we investigate a novel alternating current electrothermal (ACET) micromixer driven by a high efficiency ACET micropump. The micromixer consists of thin film asymmetric pairs of electrodes on the microgrooved channel floor and array of electrode pairs fabricated on the top wall. By connecting electrodes with AC voltage, ACET forces are induced. Asymmetric microgrooved electrodes force the fluids along the channel, while lateral vortex pairs are generated by symmetric electrode pairs located on the top wall. Waviness of the floor increases contact area between two confluent streams within a narrow confinement. An active mixer operates as a semi active semi passive mixer. Effects of various parameters are investigated in details in order to arrive at an optimal configuration that provides for efficient mixing as well as appreciable transport. It is found that using a specific design, uniform and homogeneous mixing quality with mixing efficiency of 97.25% and flow rate of 1.794μm2/ min per unit width of the channel can be achieved.


Epigenetic silencing of microRNA-218 via EZH2-mediated H3K27 trimethylation is involved in malignant transformation of HBE cells induced by cigarette smoke extract.
Abnormal expression of miRNAs has been implicated in the pathogenesis of human lung cancers, most of which are attributable to cigarette smoke. The mechanisms of action, however, remain obscure. Here, we report that there are decreased expression of miR-218 and increased expression of EZH2 and H3K27me3 during cigarette smoke extract (CSE)-induced transformation of human bronchial epithelial (HBE) cells. Depletion of EZH2 by siRNA or by the EZH2 inhibitor, 3-deazaneplanocin A, attenuated CSE-induced decreases of miR-218 levels and increases of H3K27me3, which epigenetically controls gene transcription, and BMI1, an oncogene. Furthermore, ChIP assays demonstrated that EZH2 and H3K27me3 are enriched at the miR-218-1 promoter in HBE cells exposed to CSE, indicating that EZH2 mediates epigenetic silencing of miR-218 via histone methylation. In addition, miR-218 directly targeted BMI1, through which miR-218 ablates cancer stem cells (CSCs) self-renewal in transformed HBE cells. In CSE-transformed HBE cells, the protein level of Oct-4 and mRNA levels of CD133 and CD44, indicators of the acquisition of CSC-like properties, were reduced by over-expression of miR-218, and over-expression of miR-218 decreased the malignancy of transformed HBE cells. Thus, we conclude that epigenetic silencing of miR-218 via EZH2-mediated H3K27 trimethylation is involved in the acquisition of CSC-like properties and malignant transformation of HBE cells induced by CSE and thereby contributes to the carcinogenesis of cigarette smoke.


HYPOTHYROXINAEMIA AND BRAIN DEVELOPMENT.
The aim of this review is to indicate the current position on the role of thyroxine (T4) and fetal brain development with particular relevance to the human situation. Adequate maternal iodine nutrition and maternal circulating thyroxine (T4) concentrations are essential to ensure optimum T4 placental passage which in turn will ensure transport of T4 into fetal brain cells. These processes are discussed and the role of thyroid hormone transporters is considered. The emphasis on isolated maternal hypothyroxinaemia (IH) as an important factor affecting brain development is discussed from the animal experimental point of view as well as in the clinical setting. There is evidence of neurocognitive impairment as assessed by different modalities in children up to the age of 8 years and some suggestion of increased psychiatric disorder in older persons whose mothers had IH during gestation. Although international guidelines have not in general recommended thyroxine therapy for IH the recent demonstration of adverse obstetric outcomes in women with isolated maternal hypothyroxinaemia may warrant a revision of this strategy.


A high resolution scanning electron microscope for in situ investigation of swift heavy ion induced modification of solid surfaces.
A new in situ high resolution electron microscope (HRSEM) setup has been designed and integrated into the materials science beamline (M-branch) of the universal linear accelerator at Gesellschaft für Schwerionenforschung, Darmstadt for in situ investigations of swift heavy ion induced surface modifications. Special ports for in situ experiments are installed at the HRSEM chamber to connect it to the high vacuum beamline, which is equipped with suitable beam control and shaping devices. In order to demonstrate the feasibility and power of this new instrument, first experiments were performed on a 50-nm-thick Fe(2)O(3) film on Si substrate, which exhibited submicrometer size holes due to irradiation induced dewetting in a previous experiment. We have demonstrated that with our new instrument, the development of individual dewetting holes as a function of the ion fluence can be investigated and even the interaction between them can be studied. To illustrate pattern formation during grazing incidence, 3.6 MeV/u (131)Xe ion irradiation was carried out on a 25-nm-thick NiO film on SiO(2)/Si at a tilt angle of 75 degrees. The SEM image sequence recorded during the experiment reveals the development of a lamellaelike structure also seen before in ex situ experiments. With our new in situ setup, however, we are able to not only investigate the overall average pattern formation, but also to track the formation and decay of individual linking structures, which would be hardly possible in a standard ex situ experiment.


Identifying CpG sites with different differential methylation frequencies in colorectal cancer tissues based on individualized differential methylation analysis.
A big challenge to clinical diagnosis and therapy of colorectal cancer (CRC) is its extreme heterogeneity, and thus it would be of special importance if we could find common biomarkers besides subtype-specific biomarkers for CRC. Here, with DNA methylation data produced by different laboratories, we firstly revealed that the relative methylation-level orderings (RMOs) of CpG sites within colorectal normal tissues are highly stable but widely disrupted in the CRC tissues. This finding provides the basis for using the RankComp algorithm to identify differentially methylated (DM) CpG sites in every individual CRC sample through comparing the RMOs within the individual sample with the stable RMOs predetermined in normal tissues. For 75 CRC samples, RankComp detected averagely 4,062 DM CpG sites per sample and reached an average precision of 91.34% in terms that the hypermethylation or hypomethylation states of the DM CpG sites detected for each cancer sample were consistent with the observed differences between this cancer sample and its paired adjacent normal sample. Finally, we applied RankComp to identify DM CpG sites for each of the 268 CRC samples from The Cancer Genome Atlas and found 26 and 143 genes whose promoter regions included CpG sites that were hypermethylated and hypomethylated, respectively, in more than 95% of the 268 CRC samples. Individualized pathway analysis identified six pathways that were significantly enriched with DM genes in more than 90% of the CRC tissues. These universal DNA methylation biomarkers could be important diagnostic makers and therapy targets for CRC.


Service evaluation of an educational intervention to improve sexual health services in primary care implemented using a step-wedge design: analysis of chlamydia testing and diagnosis rate changes.
Providing sexual health services in primary care is an essential step towards universal provision. However they are not offered consistently. We conducted a national pilot of an educational intervention to improve staff's skills and confidence to increase chlamydia testing rates and provide condoms with contraceptive information plus HIV testing according to national guidelines, known as 3Cs&HIV. The effectiveness of the pilot on chlamydia testing and diagnosis rates in general practice was evaluated. The pilot was implemented using a step-wedge design over three phases during 2013 and 2014 in England. The intervention combined educational workshops with posters, testing performance feedback and continuous support. Chlamydia testing and diagnosis rates in participating general practices during the control and intervention periods were compared adjusting for seasonal trends in chlamydia testing and differences in practice size. Intervention effect modification was assessed for the following general practice characteristics: chlamydia testing rate compared to national median, number of general practice staff employed, payment for chlamydia screening, practice urban/rurality classification, and proximity to sexual health clinics. The 460 participating practices conducted 26,021 tests in the control period and 18,797 tests during the intervention period. Intention-to-treat analysis showed no change in the unadjusted median tests and diagnoses per month per practice after receiving training: 2.7 vs 2.7; 0.1 vs 0.1. Multivariable negative binomial regression analysis found no significant change in overall testing or diagnoses post-intervention (incidence rate ratio (IRR) 1.01, 95 % confidence interval (CI) 0.96-1.07, P = 0.72; 0.98 CI 0.84-1.15, P = 0.84, respectively). Stratified analysis showed testing increased significantly in practices where payments were in place prior to the intervention (IRR 2.12 CI 1.41-3.18, P < 0.001) and in practices with 6-15 staff (6-10 GPs IRR 1.35 (1.07-1.71), P = 0.012; 11-15 GPs IRR 1.37 (1.09-1.73), P = 0.007). This national pilot of short educational training sessions found no overall effect on chlamydia testing in primary care. However, in certain sub-groups chlamydia testing rates increased due to the intervention. This demonstrates the importance of piloting and evaluating any service improvement intervention to assess the impact before widespread implementation, and the need for detailed understanding of local services in order to select effective interventions.


Attitude of dental hygienists, general practitioners and periodontists towards preventive oral care: an exploratory study.
Promoting oral hygiene and stimulating patient's responsibility for his/her personal health remain challenging objectives. The presence of dental hygienists has led to delegation of preventive tasks. However, in some countries, such as Belgium, this profession is not yet legalized. The aim of this exploratory study was to compare the attitude towards oral-hygiene instructions and patient motivational actions by dental hygienists and by general practitioners/periodontists in a context without dental hygienists. A questionnaire on demographics (six items), oral-hygiene instructions (eight items) and patient motivational actions (six items) was distributed to 241 Dutch dental hygienists, 692 general practitioners and 32 periodontists in Flanders/Belgium. Statistical analysis included Fisher's exact-test, Pearson's chi-square test and multiple (multinomial) logistic regression analysis to observe the influence of profession, age, workload, practice area and chair-assistance. Significant variance was found between general practitioners and dental hygienists (in 13 of 14 items), between general practitioners and periodontists (in nine of 14 items) and between dental hygienists and periodontists (in five of 14 items). In addition to qualification, chair-assistance was also identified as affecting the attitude towards preventive oral care. The present study identified divergence in the application of, and experienced barriers and opinions about, oral-hygiene instructions and patient motivational actions between dental hygienists and general practitioners/periodontists in a context without dental hygienists. In response to the barriers reported it is suggested that preventive oriented care may benefit from the deployment of dental hygienists to increase access to qualified preventive oral care.


Sphingosine 1-phosphate receptors regulate TLR4-induced CXCL5 release from astrocytes and microglia.
Sphingosine 1-phosphate receptors (S1PR) are G protein-coupled and compose a family with five subtypes, S1P1R-S1P5R. The drug Gilenya® (Novartis, Basel, Switzerland) (Fingolimod; FTY720) targets S1PRs and was the first oral therapy for patients with relapsing-remitting multiple sclerosis (MS). The phosphorylated form of FTY720 (pFTY720) binds S1PRs causing initial agonism, then subsequent receptor internalization and functional antagonism. Internalization of S1P1R attenuates sphingosine 1-phosphate (S1P)-mediated egress of lymphocytes from lymph nodes, limiting aberrant immune function in MS. pFTY720 also exerts direct actions on neurons and glial cells which express S1PRs. In this study, we investigated the regulation of pro-inflammatory chemokine release by S1PRs in enriched astrocytes and microglial cultures. Astrocytes and microglia were stimulated with lipopolysaccharide (LPS) and increases in C-X-C motif chemokine 5 (CXCL5), also known as LIX (lipopolysaccharide-induced CXC chemokine) expression were quantified. Results showed that pFTY720 attenuated LPS-induced CXCL5 (LIX) protein release from astrocytes, as did the S1P1R selective agonist, SEW2871. In addition, pFTY720 blocked messenger ribonucleic acid (mRNA) transcription of the chemokines, (i) CXCL5/LIX, (ii) C-X-C motif chemokine 10 (CXCL10) also known as interferon gamma-induced protein 10 (IP10) and (iii) chemokine (C-C motif) ligand 2 (CCL2) also known as monocyte chemoattractant protein 1 (MCP1). Interestingly, inhibition of sphingosine kinase attenuated LPS-induced increases in mRNA levels of all three chemokines, suggesting that LPS-TLR4 (Toll-like receptor 4) signalling may enhance chemokine expression via S1P-S1PR transactivation. Lastly, these observations were not limited to astrocytes since we also found that pFTY720 attenuated LPS-induced release of CXCL5 from microglia. These data highlight a role for S1PR signalling in regulating the levels of chemokines in glial cells and support the notion that pFTY720 efficacy in multiple sclerosis may involve the direct modulation of astrocytes and microglia.


Synthesis, binding properties, and 18F labeling of fluorocarazolol, a high-affinity beta-adrenergic receptor antagonist.
New beta-adrenergic receptor antagonists, 2-(R)-(+)- and 2-(S)-(-)-1-(9H-carbazol-4-yl-oxy)-3-[[1-(fluoromethyl)ethyl]amino ]-2-propanol ((S)- and (R)-fluorocarazolols), were labeled with fluorine-18 at the no-carrier-added level by reductive alkylation of desisopropylcarazolol (4-(2-hydroxy-3-amino-1-propoxy)carbazole) with [18F]fluoroacetone. The latter was prepared by nucleophilic substitution of fluoride on acetol tosylate and may serve as a useful synthetic precursor for other radiotracers. The radiochemical yield of [18F]fluorocarazolol (500-1200 Ci/mmol) from [18F]fluoride was 40 +/- 10% at the end of the 45 min synthesis. Chiral HPLC showed > 99% enantiomeric purity of 2-(S)- and 2-(R)-[18F]fluorocarazolols. The log P of fluorocarazolol was 2.2 at pH 7.4. The in vitro KD values of (S)- and (R)-fluorocarazolol for the beta-adrenergic receptor were measured in a rat heart preparation to be KD = 68 and 1128 pM, respectively. Biodistribution experiments in mice demonstrated specific beta-adrenergic receptor binding of (S)-[18F]fluorocarazolol. (R)-[18F]fluorocarazolol showed no observable specific binding to beta-receptors in vivo. The uptake of (R)-[18F]fluorocarazolol may therefore be used as an estimation of nonspecific binding. Positron emission tomography images of pigs showed receptor-specific uptake of (S)-[18F]fluorocarazolol in the heart and lung. Washout of dissociated ligand from the tissue was observed only after 70 min postinjection. The maximum ratio of specific to nonspecific uptake in pig heart and lung was ca. 10 at 150 min postinjection. Observed levels of fluorocarazolol metabolites in mouse and pig blood were relatively low and remained fairly constant during the period from 10 to 180 min postinjection. These results indicate that (S)-(-)-[18F]fluorocarazolol is of interest for use as a radiopharmaceutical for estimation of beta-adrenergic receptors with positron tomography.


Novel mechanism whereby nuclear factor kappaB mediates DNA damage repair through regulation of O(6)-methylguanine-DNA-methyltransferase.
O(6)-Methylguanine-DNA-methyltransferase (MGMT) and nuclear factor kappaB (NF-kappaB) are two key effectors associated with the development of resistance to alkylating agent-based chemotherapy. This prompted us to hypothesize that NF-kappaB might be involved in MGMT regulation. Consistent with this hypothesis, we have discovered two putative NF-kappaB binding sites within the MGMT promoter region and showed a specific and direct interaction of NF-kappaB at each of these sites. Forced expression of the NF-kappaB subunit p65 in HEK293 cells induced an increase in MGMT expression whereas addition of the NF-kappaB super repressor DeltaNIkappaB completely abrogated the induction. We also found a significant correlation between the extent of NF-kappaB activation and MGMT expression in the glioma cell lines and the human glial tumors tested and showed that it was independent of MGMT promoter methylation. Our results are of potential clinical significance because we show that cell lines with ectopic p65 or high constitutive NF-kappaB activity are less sensitive to nitrosourea treatment and that suppression of MGMT activity with O(6)-benzylguanine completely abolishes the chemoresistance acquired by NF-kappaB. The findings of our study strongly suggest that NF-kappaB plays a major role in MGMT regulation and that MGMT is most probably the major player in NF-kappaB-mediated chemoresistance to alkylating agents.


Strongly magnetic iron nanoparticles improve the diagnosis of small tumours in the reticuloendothelial system by magnetic resonance imaging.
Despite advances in non-invasive medical imaging, accurate nodal staging of malignancy continues to rely on surgery. Superparamagnetic iron oxide nanoparticles (IONP) with lymphotropic qualities have shown some promise as contrast agents for MRI of the lymph nodes, but recent large-scale studies failed to show consistent detection of tumours below 5 mm. Herein we compare imaging of splenic and lymph node tissue using iron/iron oxide core/shell nanoparticles (Fe NP) that have superior magnetic qualities to IONP, to determine whether improved negative contrast in T(2)-weighted MRI can enhance the diagnosis of small tumours in the reticuloendothelial system. To provide an in vivo pre-clinical model of human lymph node micrometastases, breast cancer cells were injected into the spleens of mice, providing localised areas of tumour growth. MR images of groups of tumour-bearing and sham-treated animals were generated using a 1.5 T imaging system and analysed by two independent, blinded radiologists. Fe NP improved the sensitivity and specificity of MRI when compared to IONP, enabling accurate detection of tumours as small as 1-3 mm. The use of Fe NP as contrast agents have the potential to improve the diagnostic accuracy of MRI in cancer patients, leading to more rapid and effective treatment.


Homing genes, cell therapy and stroke.
Stem cell therapies, such as bone marrow transplantation, are a promising strategy for the treatment of stroke. Bone marrow-derived stem cells (BMSCs) including both hematopoietic and mesenchymal stem cells (HSCs and MSCs) can exhibit tremendous cellular differentiation in numerous organs. BMSCs may also promote structural and functional repair in several organs such as the heart, liver, brain, and skeletal muscle via stem cell plasticity. Interestingly, ischemia is known to induce mobilization of BMSCs in both animal models and humans. The tissue injury is "sensed" by the stem cells and they migrate to the site of damage and undergo differentiation. The plasticity, differentiation, and migratory functions of BMSCs in a given tissue are dependent on the specific signals present in the local micro-environment of the damaged tissue. Therefore, the ischemic micro-environment has critical patho-biological functions that are essential for the seeding, expansion, survival, renewal, growth and differentiation of BMSCs in damaged brain remodeling. Recent studies have identified the specific molecular signals, such as SDF-1/CXCR4, required for the interaction of BMSCs and damaged host tissues. Understanding the exact molecular basis of stem cell plasticity in relation to local ischemic signals could offer new insights to permit better management of stroke and other ischemic disorders. The aim of this review is to summarize recent studies into how BMSCs reach, recognize, and function in cerebral ischemic tissues, with particular regard to phenotypical reprogramming of stem cells, or "stem cell plasticity".


Estimation of drug receptor occupancy when non-displaceable binding differs between brain regions – extending the simplified reference tissue model.
The simplified reference tissue model (SRTM) is used for estimation of receptor occupancy assuming that the non-displaceable binding in the reference region is identical to the brain regions of interest. The aim of this work was to extend the SRTM to also account for inter-regional differences in non-displaceable concentrations, and to investigate if this model allowed estimation of receptor occupancy using white matter as reference. It was also investigated if an apparent higher affinity in caudate compared with other brain regions, could be better explained by a difference in the extent of non-displaceable binding. The analysis was based on a PET study in six healthy volunteers using the 5-HT1B receptor radioligand [(11)C]-AZ10419369. The radioligand was given intravenously as a tracer dose alone and following different oral doses of the 5-HT1B receptor antagonist AZD3783. Non-linear mixed effects models were developed where differences between regions in non-specific concentrations were accounted for. The properties of the models were also evaluated by means of simulation studies. The estimate (95% CI) of Ki(PL) was 10.2 ng ml(-1) (5.4, 15) and 10.4 ng ml(-1) (8.1, 13.6) based on the extended SRTM with white matter as reference and based on the SRTM using cerebellum as reference, respectively. The estimate (95% CI) of Ki(PL) for caudate relative to other brain regions was 55% (48, 62%). The extended SRTM allows consideration of white matter as reference region when no suitable grey matter region exists. AZD3783 affinity appears to be higher in the caudate compared with other brain regions.


Additive Manufacturing for Surgical Planning of Mandibular Fracture.
Currently, imaging techniques such as Computed Tomography with three-dimensional reconstruction (3D) and Magnetic Resonances are being routinely used in pre-surgical planning in all fields of medicine. Nowadays, virtual three-dimensional images, commonly displayed on two-dimensional surfaces, such as the computer screen, can be used to produce rapidly prototyped models, with excellent dimensional accuracy and fine reproduction of anatomical structures, providing professionals with the ability to use the biomodel in planning and simulating medical and dental procedures (oral and maxillofacial surgery, making individualized facial implants and prostheses, measurements and previous adaptations of prefabricated fixation plates), thus contributing to considerable reductions in surgical time and consequently the duration of anesthesia, minimizing infection risks and reducing hospital costs. In this report, we describe a case of surgical planning and treatment of bilateral atrophic mandibular fracture, in which, for surgical planning, authors used Rapid Prototyping as an adjunct tool, considering the advantages already outlined.


The effect of HAART on humoral immune response in primary HIV-1 infected patients.
The effect of Highly Active Antiretroviral Therapy (HAART) on binding and neutralizing antibody responses to human immunodeficiency virus type-1 (HIV-1) during primary and chronic infection was investigated. Seven patients HAART treated during primary infection, six HAART treated during chronic infection and five patients treated only with ZVD (Zidovudine) were analysed. HAART inhibited the development of anti env antibodies during primary infection. Administering HAART during primary infection usually did not substantially affect the development of weak neutralizing antibody responses against autologous virus. However, we demonstrated that very early treatment, during seroconversion, induce in some cases, a strong neutralizing antibodies against autologous virus. These results may be relevant for understanding how HAART may elicit a strong protective responses and may be useful in developing new strategies designed to achieve a long term control of the HIV infection.


[Neuro-oculo-cutaneous syndrome with multiple sebaceous nevi. Presentation of a case].
The authors present a case of multiple sebaceous naevi associated with severe mental retardation and ocular lesions in a six month-old female. Several elevated yellowish lesions were present in the skin of the face and an extensive atrophic lesion was observed in the scalp. Histopathological examination of the elevated yellowish plaque disclosed hyperplasia of mature sebaceous glands. In the linear lesion it was observed hyperplasia of hypotrophic sebaceous glands. Otherwise in the atrophic lesion of the scalp atrophy of the dermis associated with marked hypoplasia of the skin appendages was observed. The authors believe that the different cutaneous pathological aspects of this syndrome are related to the clinical morphology of the lesions and not to the age of the patient. The ophthalmological examination revealed bilateral optic atrophy, bilateral choristomas of the conjuntiva and a coloboma of the choroid and retina in the right eye. A review of the literature revealed eleven cases of sebaceous naevi associated with neurologic and/or ocular manifestations but only three cases of the neuro-oculo-cutaneous syndrome. The present case probably represents the fourth case of this syndrome in the literature.


Photoreconfigurable supramolecular nanotube.
A photoreconfigurable bionanotube was developed by Mg(2+)-induced supramolecular polymerization using GroELSP, a mutant barrel-shaped chaperonin protein bearing multiple photochromic spiropyran (SP) units at its apical domains. Upon exposure to UV light, the nonionic SP units isomerize into ionic merocyanine (MC) to afford GroELMC, which is capable of polymerizing with MgCl2. The resultant nanotube (NT) is stable as a result of multiple MC···Mg(2+)···MC bridges but readily breaks up into short NTs, including monomeric GroELSP, by the reverse (MC → SP) isomerization mediated by visible light. When this scission mixture is exposed to UV light, long NTs are reconfigured. A Förster resonance energy transfer (FRET) study revealed that NTs in the dark maintain their sequential integrity. However, when exposed to visible and UV light successively, the NTs lose their sequential memory as a result of intertubular reshuffling of the constituent GroELMC units.


Human brucellosis in Maghreb: existence of a lineage related to socio-historical connections with Europe.
Despite control/eradication programs, brucellosis, major worldwide zoonosis due to the Brucella genus, is endemic in Northern Africa and remains a major public health problem in the Maghreb region (Algeria/Morocco/Tunisia). Brucella melitensis biovar 3 is mostly involved in human infections and infects mainly small ruminants. Human and animal brucellosis occurrence in the Maghreb seems still underestimated and its epidemiological situation remains hazy. This study summarizes official data, regarding Brucella melitensis infections in Algeria, from 1989 to 2012, with the purpose to provide appropriate insights concerning the epidemiological situation of human and small ruminant brucellosis in Maghreb. Algeria and Europe are closely linked for historical and economical reasons. These historical connections raise the question of their possible impact on the genetic variability of Brucella strains circulating in the Maghreb. Other purpose of this study was to assess the genetic diversity among Maghreb B. melitensis biovar 3 strains, and to investigate their possible epidemiological relationship with European strains, especially with French strains. A total of 90 B. melitensis biovar 3 Maghreb strains isolated over a 25 year-period (1989-2014), mainly from humans, were analysed by MLVA-16. The obtained results were compared with genotypes of European B. melitensis biovar 3 strains. Molecular assays showed that Algerian strains were mainly distributed into two distinct clusters, one Algerian cluster related to European sub-cluster. These results led to suggest the existence of a lineage resulting from socio-historical connections between Algeria and Europe that might have evolved distinctly from the Maghreb autochthonous group. This study provides insights regarding the epidemiological situation of human brucellosis in the Maghreb and is the first molecular investigation regarding B. melitensis biovar 3 strains circulating in the Maghreb.


The effect of contingent music to increase non-nutritive sucking of premature infants.
This study assessed music as reinforcement for non-nutritive sucking of 12 premature infants born at an average gestation of 29.3 weeks and an average birthweight of 1111.9 g. At the time of the study, the infants' average post conception age was 35.5 weeks, and their average weight was 1747.3 g. A pacifier was fitted with a pressure transducer so that a sufficient suck activated frequency and duration signals as well as 10 seconds of recorded music consisting of lullabies sung by female vocalists. A 14-minute ABAB study design included a silence baseline for 2 minutes, 5 minutes of contingent music, 2 minutes of silence, and 5 minutes of contingent music. Frequency data were recorded for each 5-second interval in which the duration light was activated for at least 3 seconds. Results demonstrated that sucking rates during the periods of contingent music were 2.43 times greater than baseline (silence) sucking rates. In this study, music contributed significantly to the development of non-nutritive sucking of premature infants.


Differences in reported linguistic thermal sensation between Bangla and Japanese speakers.
Thermal sensation is a fundamental variable used to determine thermal comfort and is most frequently evaluated through the use of subjective reports in the field of environmental physiology. However, there has been little study of the relationship between the semantics of the words used to describe thermal sensation and the climatic background. The present study investigates the linguistic differences in thermal reports from native speakers of Bangla and Japanese. A total of 1141 university students (932 in Bangladesh and 209 in Japan) responded to a questionnaire survey consisting of 20 questions. Group differences between Bangladeshi and Japanese respondents were then tested with a chi-square test in a crosstab analysis using SPSS (version 21). For the Bangla-speaking respondents, the closest feeling of thermal comfort was "neutral" (66.6%) followed by "slightly cool" (10.2%), "slightly cold" (6.0%), "slightly hot" (4.1%), and "cold" (3.8%). For the Japanese respondents, the closest feeling of thermal comfort was "cool" (38.3%) followed by "slightly cool" (20.4%), "neutral" (14.6%), "slightly warm" (13.1%), and "warm" (10.7%). Of the Bangladeshi respondents, 37.7% reported that they were sensitive to cold weather and 18.1% reported that they were sensitive to hot weather. Of the Japanese respondents, 20.6% reported that they were sensitive to cold weather and 29.2% reported that they were sensitive to hot weather. Of the Bangladeshi respondents, 51.4% chose "higher than 29 °C" as hot weather and 38.7% of the Japanese respondents chose "higher than 32 °C" as hot weather. In the case of cold weather, 43.1% of the Bangladeshi respondents selected "lower than 15 °C" as cold weather and 53.4% of the Japanese respondents selected "lower than 10 °C" as cold weather. Most of the Bangla-speaking respondents chose "neutral" as the most comfortable temperature, and most of the Japanese respondents chose "cool." Most of the Bangladeshi respondents reported that they were sensitive to "cold temperatures," but most of the Japanese respondents reported that they were sensitive to "hot temperatures."


Characterization of postsynaptic alpha-adrenoceptors in the isolated rat tail artery.
The main purpose was to assess the existence of alpha 2-adrenoceptors in isolated segments of the proximal portion of the rat tail artery. Adrenaline and phenylephrine evoked strong contractile responses, whereas in 22 out of 49 arteries, 10-100 microM clonidine evoked weak contractions. Guanabenz, idazoxan and yohimbine lacked contractile effects. Reserpine pretreatment did not modify the percentage of responsive arteries to clonidine nor the ineffectiveness of guanabenz. Raising the Ca2+ concentration to 2.5 mM did not alter the responsiveness to adrenaline and phenylephrine but enhanced the amplitude of the responses to clonidine. The exposure to 0.1 or 1.0 microM angiotensin did not affect the responsiveness to clonidine. Responses to clonidine were abolished by 10 nM prazosin and not affected by 1 microM idazoxan or yohimbine. Adrenaline and phenylephrine concentration-response curves were shifted rightward by prazosin and phentolamine (pA2 within 7.3-9.0) and by clonidine, guanabenz, idazoxan and yohimbine (pA2 lower than 6.5). These data suggest that the tail artery lacks postsynaptic alpha 2-adrenoceptors, that clonidine exerts partial alpha 1-agonist/antagonist effects and that guanabenz, idazoxan and yohimbine act as partial alpha 1-antagonists.


Bile acid-induced negative feedback regulation of the human ileal bile acid transporter.
Ileal expression of the apical sodium-dependent bile acid transporter (ASBT) in the rat is unaffected by bile salts, yet in the mouse it is under negative-feedback regulation. The bile acid responsiveness of human ASBT is unknown. The human ASBT promoter linked to a luciferase reporter was studied in Caco-2 cells treated with chenodeoxycholic acid (CDCA) and transfected with expression plasmids for farnesoid X-receptor (FXR), short heterodimer partner (SHP), and retinoic acid receptor/retinoid X receptor (RAR/RXR). CDCA treatment of Caco-2 cells led to a 75% reduction in steady-state ASBT messenger RNA levels and a 78% reduction in human ASBT promoter activity. A dominant negative FXR abrogated the response to CDCA. Site-directed mutagenesis of an RAR/RXR cis element in the human ASBT promoter reduced its activity by 50% and eliminated the bile acid response. Retinoic acid activated the human ASBT promoter fourfold. SHP repressed the activity of the ASBT promoter and reduced activation by retinoic acid. Antisense mediated knock-down of SHP in Caco-2 cells partially offset the bile acid mediated repression of ASBT promoter activity. In conclusion, the human ASBT is positively regulated by retinoic acid. Bile acids induce a negative feedback regulation of human ASBT via an FXR-mediated, SHP-dependent effect upon RAR/RXR activation of ASBT.