Targeted delivery of a model immunomodulator to the lymphatic system: comparison of alkyl ester versus triglyceride mimetic lipid prodrug strategies.
A lipophilic prodrug approach has been used to promote the delivery of a model immunomodulator, mycophenolic acid (MPA), to the lymphatic system after oral administration. Lymphatic transport was employed to facilitate enhanced drug uptake into lymphocytes, as recent studies demonstrate that targeted drug delivery to lymph resident lymphocytes may enhance immunomodulatory effects. Two classes of lymph-directing prodrugs were synthesised. Alkyl chain derivatives (octyl mycophenolate, MPA-C8E; octadecyl mycophenolate, MPA-C18E; and octadecyl mycophenolamide, MPA-C18AM), to promote passive partitioning into lipids in lymphatic transport pathways, and a triglyceride mimetic prodrug (1,3-dipalmitoyl-2-mycophenoloyl glycerol, 2-MPA-TG) to facilitate metabolic integration into triglyceride deacylation-reacylation pathways. Lymphatic transport, lymphocyte uptake and plasma pharmacokinetics were assessed in mesenteric lymph and carotid artery cannulated rats following intraduodenal infusion of lipid-based formulations containing MPA or MPA prodrugs. Patterns of prodrug hydrolysis in rat digestive fluid, and cellular re-esterification in vivo, were evaluated to examine the mechanisms responsible for lymphatic transport. Poor enzyme stability and low absorption appeared to limit lymphatic transport of the alkyl derivatives, although two of the three alkyl chain prodrugs - MPA-C18AM (6-fold) and MPA-C18E (13-fold) still increased lymphatic drug transport when compared to MPA. In contrast, 2-MPA-TG markedly increased lymphatic drug transport (80-fold) and drug concentrations in lymphocytes (103-fold), and this was achieved via biochemical incorporation into triglyceride deacylation-reacylation pathways. The prodrug was hydrolysed rapidly to 2-mycophenoloyl glycerol (2-MPA-MG) in the presence of rat digestive fluid, and 2-MPA-MG was subsequently re-esterified in the enterocyte with oleic acid (most likely originating from the co-administered formulation) prior to accessing the lymphatics and lymphocytes. Importantly, after administration of 2-MPA-TG, the concentrations of free MPA in the mesenteric lymph nodes were significantly enhanced (up to 28 fold) when compared to animals administered equimolar quantities of MPA, suggesting the efficient conversion of the esterified prodrug back to the pharmacologically active parent drug. The data suggest that triglyceride mimetic prodrugs have potential as a means of enhancing immunotherapy via drug targeting to lymphocytes and lymph nodes.


Virus-like particle-induced fusion from without in tissue culture cells: role of outer-layer proteins VP4 and VP7.
We recently described an assay that measures fusion from without induced in tissue culture cells by rotavirus, a nonenveloped, triple-protein-layered member of the Reoviridae family (M. M. Falconer, J. M. Gilbert, A. M. Roper, H. B. Greenberg, and J. S. Gavora, J. Virol. 69:5582-5591, 1995). The conditions required for syncytium formation are similar to those for viral penetration of the plasma membrane during the course of viral infection of host cells, as the presence of the outer-layer proteins VP4 and VP7 and the cleavage of VP4 are required. Here we present evidence that virus-like particles (VLPs) produced in Spodoptera frugiperda Sf-9 cells from recombinant baculoviruses expressing the four structural proteins of rotavirus can induce cell-cell fusion to the same extent as native rotavirus. This VLP-mediated fusion activity was dependent on trypsinization of VP4, and the strain-specific phenotype of individual VP4 molecules was retained in the syncytium assay similar to what has been seen with reassortant rotaviruses. We show that intact rotavirus and VLPs induce syncytia with cells that are permissive to rotavirus infection whereas nonpermissive cells are refractory to syncytium formation. This finding further supports our hypothesis that the syncytium assay accurately reflects very early events involved in viral infection and specifically the events related to viral entry into the cell. Our results also demonstrate that neither viral replication nor rotavirus proteins other than VP2, VP6, VP4, and VP7 are required for fusion and that both VP4 and VP7 are essential. The combination of a cell-cell fusion assay and the availability of recombinant VLPs will permit us to dissect the mechanisms of rotavirus penetration into host cells.


[Long-term results of dental implantation in military personnel].
Currently, the use of the dental implantation method for the medical service of the Armed Forces of the Russian Federation is quite new. The method of dental implantation makes it possible to eliminate defects in the dentition of military personnel and other contingents eligible for medical care in military medical institutions of the Ministry of Defense of Russia. The aim of the study is to substantiate the medical effectiveness of the use of dental implantation in military healthcare. The study was conducted on the basis of the Department of Maxillofacial Surgery and Surgical Dentistry of the Military Medical Academy named after S.M. Kirov. The study is based on the analysis of long-term results of dental implantation according to medical histories and outpatient records. The study was conducted in 3 stages: stage 1 - study of the medical histories of patients who underwent dental implantation; stage 2 - examination of patients with dental implants and analysis of long-term results; stage 3 - a patient questionnaire was conducted to determine the effectiveness of dental implantation. The structure of complications during dental implantation was revealed. Clinical examples of long-term results of prosthetics using dental implants are presented. It has been established that due to the peculiarity of military service and the remoteness of the locations of military units from the military medical organization, it is not always possible to conduct regular medical examinations and examinations of patients. To identify early symptoms of peri-implantitis, which is more often asymptomatic, preventive examinations of military personnel should be carried out 1-2 times a year. To increase the service life of an orthopedic structure with installed dental implants, regular preventive examinations of patients to identify pathological processes are important. In favor of implantation, there is a choice for some categories of military personnel. This is especially important for flight and naval personnel, since dentures installed on dental implants are the only way to preserve their professional suitability in the military accounting specialty. The presence of teeth and the absence of removable dentures is an important requirement when performing combat missions.


Delivery of home care services after discharge: what really happens.
Social workers in hospitals develop discharge plans for in-home patient care with little systematic feedback about postdischarge implementation. A telephone follow-up study of patients discharged from an urban teaching hospital in 1990 was undertaken to determine the extent to which discharge plans for home services were carried out and to identify factors associated with unsuccessful implementation. Overall, 72 percent of the patients received all, 19 percent some, and 9 percent none of the planned home care services. Great variability was found in service delivery: Registered nurse visits were the most successfully delivered type of service; 24-hour companions were the least successfully delivered service. Further, over one-third of patients experienced termination or reduction of services between discharge and the follow-up interview 21 to 28 days after discharge. Such unexpected and varied outcomes suggest the need for development of discharge follow-up programs that move beyond hospital walls to ensure that patients receive needed services.


Comparative efficiency of wheat straw and sugarcane bagasse biochar reduces the cadmium bioavailability to spinach and enhances the microbial activity in contaminated soil.
Biochar is considered a novel soil amendment for cadmium (Cd) stabilization in contaminated soils. A pot experiment was conducted to examine the efficiency of wheat straw and sugarcane bagasse induced biochar on Cd mobility in soil and its bioavailability to spinach in contaminated soil. Soil pH, Cd contents in plant tissues and microbial biomass were examined. Results showed that Cd was significantly decreased by 30.95% and 20.83% with wheat straw and sugarcane bagasse biochar at 2% application rate respectively, relative to the control. Similarly, Cd contents were decreased in plants shoots by 15.41 and 14.33%, while in roots by 48.3 and 35.54%, when wheat straw and sugarcane biochar were added at 2% application rate respectively. Moreover, soil microbial biomass was significantly increased with the application of all biochar types and their applications rates. Finally, wheat straw biochar at 2% application rate can be considered as an effective approach for Cd stabilization in contaminated soils.


Out Like a Light? The Effects of a Diurnal Husbandry Schedule on Mouse Sleep and Behavior.
Sleep disruption in humans, caused by shift work, can be detrimental to physical and behavioral health. Nocturnal laboratory mice may experience a similar disruption caused by human daytime activities, but whether this disruption affects their welfare is unknown. We used 48 mice (CD1, C57BL/6, and BALB/c of both sexes) in a factorial design to test a sleep disruption treatment, in which mice were disturbed by providing routine husbandry at either 1000 or 2200 during a 12:12-h light:dark cycle, with lights on at 0700. All mice were exposed for 1 wk to each disruption treatment, and we used a noninvasive sleep monitoring apparatus to monitor and record sleep. To determine whether providing nesting material ameliorated effects of sleep disruption, we tested 4 amounts of nesting material (3, 6, 9, or 12 g) and continuously recorded sleep in the home cage for 2 wk. C57BL/6 mice, regardless of sex or disruption timing, slept the least overall. There was a strong interaction of sex and type of mouse on sleep across 24 h. Mice slept less during the first day of the daytime disturbance than on day 6. These results suggest that disturbance timing affects sleep patterns in mice but not their overall amount of sleep and that the changes in sleep patterns vary between mouse type and sex. In addition, mice appear to both anticipate and acclimate to human activity during the day. Our welfare checks were possibly too predictable and inconsequential to induce true sleep disruption.


The crystal structures of Klebsiella pneumoniae acetolactate synthase with enzyme-bound cofactor and with an unusual intermediate.
Acetohydroxyacid synthase (AHAS) and acetolactate synthase (ALS) are thiamine diphosphate (ThDP)-dependent enzymes that catalyze the decarboxylation of pyruvate to give a cofactor-bound hydroxyethyl group, which is transferred to a second molecule of pyruvate to give 2-acetolactate. AHAS is found in plants, fungi, and bacteria, is involved in the biosynthesis of the branched-chain amino acids, and contains non-catalytic FAD. ALS is found only in some bacteria, is a catabolic enzyme required for the butanediol fermentation, and does not contain FAD. Here we report the 2.3-A crystal structure of Klebsiella pneumoniae ALS. The overall structure is similar to AHAS except for a groove that accommodates FAD in AHAS, which is filled with amino acid side chains in ALS. The ThDP cofactor has an unusual conformation that is unprecedented among the 26 known three-dimensional structures of nine ThDP-dependent enzymes, including AHAS. This conformation suggests a novel mechanism for ALS. A second structure, at 2.0 A, is described in which the enzyme is trapped halfway through the catalytic cycle so that it contains the hydroxyethyl intermediate bound to ThDP. The cofactor has a tricyclic structure that has not been observed previously in any ThDP-dependent enzyme, although similar structures are well known for free thiamine. This structure is consistent with our proposed mechanism and probably results from an intramolecular proton transfer within a tricyclic carbanion that is the true reaction intermediate. Modeling of the second molecule of pyruvate into the active site of the enzyme with the bound intermediate is consistent with the stereochemistry and specificity of ALS.


Effect of exclusive breastfeeding and complementary feeding on infant growth and morbidity.
A cohort study was conducted in the Islamic Republic of Iran between January 1997 and February 1998 to compare the growth and morbidity of 100 infants who were exclusively breastfed for 6 months and 100 who received breast milk and complementary foods between 4-6 months. Infants' feeding pattern, weight and height were assessed and recorded. There were no significant differences in infants' weight and height gain between 4 and 6 months. The rate of diarrhoea between ages 4 and 6 months was significantly lower in exclusively breastfed infants than in complementary food-fed infants (11% versus 27%) and respiratory infections were also lower (23% versus 35%). We conclude that exclusive breastfeeding is superior at least until an infant is 6 months of age.


Cone-beam breast computed tomography using ultra-fast image reconstruction with constrained, total-variation minimization for suppression of artifacts.
Compressed sensing based iterative reconstruction algorithms for computed tomography such as adaptive steepest descent-projection on convex sets (ASD-POCS) are attractive due to their applicability in incomplete datasets such as sparse-view data and can reduce radiation dose to the patients while preserving image quality. Although IR algorithms reduce image noise compared to analytical Feldkamp-Davis-Kress (FDK) algorithm, they may generate artifacts, particularly along the periphery of the object. One popular solution is to use finer image-grid followed by down-sampling. This approach is computationally intensive but may be compensated by reducing the field of view. Our proposed solution is to replace the algebraic reconstruction technique within the original ASD-POCS by ordered subsets-simultaneous algebraic reconstruction technique (OS-SART) and with initialization using FDK image. We refer to this method as Fast, Iterative, TV-Regularized, Statistical reconstruction Technique (FIRST). In this study, we investigate FIRST for cone-beam dedicated breast CT with large image matrix. The signal-difference to noise ratio (SDNR), the difference of the mean value and the variance of adipose and fibroglandular tissues for both FDK and FIRST reconstructions were determined. With FDK serving as the reference, the root-mean-square error (RMSE), bias, and the full-width at half-maximum (FWHM) of microcalcifications in two orthogonal directions were also computed. Our results suggest that FIRST is competitive to the finer image-grid method with shorter reconstruction time. Images reconstructed using the FIRST do not exhibit artifacts and outperformed FDK in terms of image noise. This suggests the potential of this approach for radiation dose reduction in cone-beam breast CT.


Vitamin D and the brain: Genomic and non-genomic actions.
1,25(OH)2D3 (vitamin D) is well-recognized as a neurosteroid that modulates multiple brain functions. A growing body of evidence indicates that vitamin D plays a pivotal role in brain development, neurotransmission, neuroprotection and immunomodulation. However, the precise molecular mechanisms by which vitamin D exerts these functions in the brain are still unclear. Vitamin D signalling occurs via the vitamin D receptor (VDR), a zinc-finger protein in the nuclear receptor superfamily. Like other nuclear steroids, vitamin D has both genomic and non-genomic actions. The transcriptional activity of vitamin D occurs via the nuclear VDR. Its faster, non-genomic actions can occur when the VDR is distributed outside the nucleus. The VDR is present in the developing and adult brain where it mediates the effects of vitamin D on brain development and function. The purpose of this review is to summarise the in vitro and in vivo work that has been conducted to characterise the genomic and non-genomic actions of vitamin D in the brain. Additionally we link these processes to functional neurochemical and behavioural outcomes. Elucidation of the precise molecular mechanisms underpinning vitamin D signalling in the brain may prove useful in understanding the role this steroid plays in brain ontogeny and function.


Glassy states in adsorbing surfactant-microgel soft nanocomposites.
Mixtures of polymer-colloid hybrids such as star polymers and microgels with non-adsorbing polymeric additives have received a lot of attention. In these materials, the interplay between entropic forces and softness is responsible for a wealth of phenomena. By contrast, binary mixtures where one component can adsorb onto the other one have been far less studied. Yet real formulations in applications often contain low molecular weight additives that can adsorb onto soft colloids. Here we study the microstructure and rheology of soft nanocomposites made of surfactants and microgels using linear and nonlinear rheology, SAXS experiments, and cryo-TEM techniques. The results are used to build a dynamical state diagram encompassing various liquid, glassy, jammed, metastable, and reentrant liquid states, which results from a subtle interplay between enthalpic, entropic, and kinetic effects. We rationalize the rheological properties of the nanocomposites in each domain of the state diagram, thus providing exquisite solutions for designing new rheology modifiers at will.


Solvent effect in the synthesis of hydrophobic drug-loaded polymer nanoparticles.
Quercetin is an abundant flavonoid in fruits, vegetables such as onion, tea leaves, cranberry, radish leaves etc. with numerous biological activities and widely used as an effective antioxidant. Its low solubility in water and chemical decomposition in intestinal environment are predicaments in delivery through dietary or oral intake. Noble polymeric nanoparticles are of particular interest today because of their applications in many areas. Polymer nanoparticles have attracted the interest of many research groups and have been utilised in an increasing number of fields such as site targeted drug delivery in cancer research during the last decades. Various techniques can be used to produce polymer nanoparticles, such as solvent evaporation, salting-out, dialysis, supercritical fluid technology etc. The choice of method depends on a number of factors, such as, particle size, particle size distribution, area of application, etc. In the present study, single emulsion-solvent evaporation technique has been utilised with two different organic solvents: acetone and chloroform/methanol to prepare quercetin loaded poly(D,L-lactide-co-glycolide) nanoparticles. According to the authors' observations acetone is a better solvent for encapsulating quercetin in polymer nanoparticles owing to its physical and chemical properties.


Impact of Vascular Hemodynamics on Aortic Stenosis Evaluation: New Insights Into the Pathophysiology of Normal Flow-Small Aortic Valve Area-Low Gradient Pattern.
About 50% of normal-flow/low-gradient patients (ie, low mean gradient [MG] or peak aortic jet velocity and small aortic valve area) have severe aortic valve calcification as measured by computed tomography. However, they are considered to have moderate aortic stenosis (AS) in current American College of Cardiology/American Heart Association guidelines. The objective was thus to evaluate the effect of hypertension and reduced arterial compliance (rAC) on MG and Vpeak measurements. Doppler-echocardiography was performed in 4 sheep with experimentally induced severe and critical AS at: (1) normal aortic pressure, (2) during hypertension, and (3) with rAC. Hypertension and rAC induced a substantial decrease in MG/Vpeak compared with normal stage (both P≤0.03) despite a stable transvalvular flow (P>0.16). Hypertension and rAC resulted in a greater reduction of MG in critical (-42%) compared with severe (-35%) AS (P˂0.0001). Comprehensive Doppler-echocardiography and computed tomography were performed in 220 AS patients (mean age: 69±13 years; MG 29±18 mm Hg) with normal flow. The population was divided in 3 groups according to the presence of hypertension and rAC. The slope of the linear association between MG/Vpeak and aortic valve calcification divided by the cross-sectional area of the aortic annulus was significantly reduced in patients with hypertension and/or rAC compared with normotensive/normal AC patients (P<0.01). Accordingly, patients with normal-flow/low-gradient and severe aortic valve calcification density were more frequent in hypertension and rAC groups compared with the normotensive/normal-AC group (16% and 12% compared with 2%; P=0.03). Hypertension and rAC are associated with a substantial reduction in MG/Vpeak for similar aortic valve calcification (ie, similar AS anatomic severity), which may lead to underestimation of AS hemodynamic severity.


Direct interaction between USF and SREBP-1c mediates synergistic activation of the fatty-acid synthase promoter.
To understand the molecular mechanisms underlying transcriptional activation of fatty-acid synthase (FAS), we examined the relationship between upstream stimulatory factor (USF) and SREBP-1c, two transcription factors that we have shown previously to be critical for FAS induction by feeding/insulin. Here, by using a combination of tandem affinity purification and coimmunoprecipitation, we demonstrate, for the first time, that USF and SREBP-1 interact in vitro and in vivo. Glutathione S-transferase pulldown experiments with various USF and sterol regulatory element-binding protein (SREBP) deletion constructs indicate that the basic helix-loop-helix domain of USF interacts directly with the basic helix-loop-helix and an N-terminal region of SREBP-1c. Furthermore, cotransfection of USF and SREBP-1c with an FAS promoter-luciferase reporter construct in Drosophila SL2 cells results in highly synergistic activation of the FAS promoter. We also show similar cooperative activation of the mitochondrial glycerol-3-phosphate acyltransferase promoter by USF and SREBP-1c. Chromatin immunoprecipitation analysis of mouse liver demonstrates that USF binds constitutively to the mitochondrial glycerol 3-phosphate acyltransferase promoter during fasting/refeeding in vivo, whereas binding of SREBP-1 is observed only during refeeding, in a manner identical to that of the FAS promoter. In addition, we show that the synergy we have observed depends on the activation domains of both proteins and that mutated USF or SREBP lacking the N-terminal activation domain could inhibit the transactivation of the other. Closely positioned E-boxes and sterol regulatory elements found in the promoters of several lipogenic genes suggest a common mechanism of induction by feeding/insulin.


Comparative biomembrane permeation of tacrine using Yucatan minipigs and domestic pigs as the animal model.
Tacrine (THA), a centrally acting acetylcholine-esterase inhibitor, is presently administered perorally for the treatment of Alzheimer's disease (AD). However, its low bioavailablity (i.e., 17%) and short half-life (2-4 h) demand the search for alternative routes of administration. The primary objective of this study was to assess the potential of absorptive mucosae and skin as routes for improving the systemic delivery of THA. The Yucatan minipig, which has been used increasingly in biomedical research as a useful model for humans, and the domestic pig, which is available at low cost, were evaluated for their suitability as animal model. Permeation kinetics of THA across various absorptive mucosae (nasal, buccal, sublingual, and rectal) of both species of swine were studied in the hydrodynamically well-calibrated Valia-Chien permeation cells. For comparison, permeation through various intestinal segments (duodenum, jejunum, and ileum) was also measured. Results indicated that both species display similar permeation characteristics. However, the data obtained for the domestic pigs shows lower intra- and inter-animal variabilities than that of the Yucatan minipigs. The nasal mucosa was found to have the highest permeability, while the buccal mucosa had the lowest among the absorptive mucosae. The intrinsic permeabilities and diffusivity of THA across the four absorptive mucosae were not significantly different between species but lower than that for the intestinal segments for both species. Using dorsal skin as the model, the skin permeation of THA was also investigated and the results indicated that the domestic swine has a significantly higher skin permeability than the Yucatan minipig, with more than a 2-fold difference in intrinsic permeabilities. The intrinsic permeability, partition coefficient, and diffusivity for domestic pig skin are very similar to that for human cadaver skin. Considering the potential of bypassing the hepatic "first-pass" elimination, the absorptive mucosae may be useful routes for systemic delivery of THA to achieve improved bioavailability. With additional advantages of lower variability, ease of membrane excision, good accessibility, and lower cost, it is concluded that the domestic swine is a better animal model than the Yucatan minipig for preclinical studies on the systemic delivery of tacrine.


Endothelial nitric oxide production during in vitro simulation of external limb compression.
External pneumatic compression (EPC) is effective in preventing deep vein thrombosis (DVT) and is thought to alter endothelial thromboresistant properties. We investigated the effect of EPC on changes in nitric oxide (NO), a critical mediator in the regulation of vasomotor and platelet function. An in vitro cell culture system was developed to simulate flow and vessel collapse conditions under EPC. Human umbilical vein endothelial cells were cultured and subjected to tube compression (C), pulsatile flow (F), or a combination of the two (FC). NO production and endothelial nitric oxide synthase (eNOS) mRNA expression were measured. The data demonstrate that in the F and FC groups, there is a rapid release of NO followed by a sustained increase. NO production levels in the F and FC groups were almost identical, whereas the C group produced the same low amount of NO as the control group. Conditions F and FC also upregulate eNOS mRNA expression by a factor of 2.08 +/- 0.25 and 2.11 +/- 0.21, respectively, at 6 h. Experiments with different modes of EPC show that NO production and eNOS mRNA expression respond to different time cycles of compression. These results implicate enhanced NO release as a potentially important factor in the prevention of DVT.


Subcellular distribution of daunorubicin in P-glycoprotein-positive and -negative drug-resistant cell lines using laser-assisted confocal microscopy.
Four well defined multidrug-resistant cell lines and their drug-sensitive counterparts were examined for intracellular distribution of daunorubicin (DNR) by laser-assisted confocal fluorescence microscopy: P-glycoprotein-negative HL-60/AR cells, and P-glycoprotein-positive P388/ADR, KBV-1, and MCF-7/ADR cells. Both drug sensitive cell lines (HL-60/S, P388/S, KB3-1, and MCF-7/S) and drug-resistant cell lines (HL-60/AR, P388/ADR, KBV-1, and MCF-7/ADR) exposed to DNR showed a similar rapid distribution of drug from the plasma membrane to the perinuclear region within the first 2 min. From 2-10 min, the drug sensitive HL-60/S, P388/S, and MCF-7/S cells redistributed drug to the nucleus and to the cytoplasm in a diffuse pattern. In contrast, drug-resistant HL-60/AR, P388/ADR, and MCF-7/ADR redistributed DNR from the perinuclear region into vesicles distinct from nuclear structures, thereby assuming a "punctate" pattern. This latter redistribution could be inhibited by glucose deprivation (indicating energy dependence), or by lowering the temperature of the medium below 18 degrees C. The differences in distribution between sensitive and resistant cells did not appear to be a function of intracellular DNR content, nor the result of drug cytotoxicity. Drug-sensitive KB3-1 and -resistant KBV-1 cells did not fully follow this pattern in that they demonstrated an intracellular DNR distribution intermediate between HL-60/S and HL-60/AR cells with both "punctate" and nuclear/cytoplasmic uptake sometimes in the same cell. These data indicate that the intracellular distribution of DNR is an important determinant of drug resistance regardless of the overexpression of P-glycoprotein. The intracellular movement of drug requires the presence of glucose and a temperature above 18 degrees C, implicating energy-dependent processes and vesicle fusion in the distribution process. This intracellular transport of DNR away from the nucleus in multidrug-resistant cells may protect putative cell targets such as DNA against drug toxicity.


A Review of Extended and Continuous Infusion Beta-Lactams in Pediatric Patients.
Intravenous beta-lactam antibiotics are the most prescribed antibiotic class in US hospitalized patients of all ages; therefore, optimizing their dosing is crucial. Bactericidal killing is best predicted by the time in which beta-lactam drug concentrations are maintained above the organism's minimum inhibitory concentration (MIC), rather than achievement of a high peak concentration. As such, administration of beta-lactam antibiotics via extended or continuous infusions over a minimum of 3 hours, rather than standard infusions over approximately 30 minutes, has been associated with improved achievement of pharmacodynamic targets and improved clinical outcomes in adult medical literature. This review summarizes the pediatric medical literature. Applicable studies include pharmacodynamic models, case series, retrospective analyses, and prospective studies on the use of extended infusion and continuous infusion penicillins, cephalosporins, carbapenems, and monobactams in neonates, infants, children, and adolescents. Specialized patient populations with unique pharmacokinetics and high-risk infections (neonates, critically ill, febrile neutropenia, cystic fibrosis) are also reviewed. While more studies are needed to confirm prospective clinical outcomes, the current body of evidence suggests extended and continuous infusions of beta-lactam antibiotics are well tolerated in children and improve achievement of pharmacokineticpharmacodynamic targets with similar or superior clinical outcomes, particularly in infections associated with high MICs.


Long term data of endovascularly treated patients with severe and complex aortoiliac occlusive disease.
This paper presents the recent data of the largest series (20 patients) of endovascularly treated patients and the first long term data of 9 patients with severe aortoiliac occlusive disease. Between 2003 and 2012, 20 consecutive patients (14 men; 70 %) with Leriche syndrome underwent recanalization with solely endovascular means at our centre. The treatment strategy comprised the antegrade (transbrachial) recanalization of the occluded segments followed by retrograde (transfemoral) angioplasty with selective stent placement in the infrarenal aorta and primary stent placement in the iliac arteries. Before discharge, after 30 days and every year after the procedure, a clinical, as well as a duplex ultrasonographic examination including measurement of the ankle-brachial index was done. Bilateral success was achieved in 17 patients (85%). Unilateral success was achieved in three patients (15%). In one patient (5%) an early reocclusion of the stented segments occurred, necessitating bypass grafting. In nine patients long term data were evaluated. Here, the ankle brachial index (ABI) significantly increased (0.85 ± 0.15 vs. 0.51 ± 0.11 at baseline; P=0.002). Compared to baseline, the difference in the distribution of Rutherford category and the improvement of walking capacity were statistically significant (P=0.0006, P=0.01, respectively). This study shows the feasibility of solely endovascular management of severe aortoiliac occlusive disease with a high rate of success and low rate of complications. Significant clinical improvement of patients in long term follow up makes the endovascular approach a viable alternative to open surgery.


Polymer physics indicates chromatin folding variability across single-cells results from state degeneracy in phase separation.
The spatial organization of chromosomes has key functional roles, yet how chromosomes fold remains poorly understood at the single-molecule level. Here, we employ models of polymer physics to investigate DNA loci in human HCT116 and IMR90 wild-type and cohesin depleted cells. Model predictions on single-molecule structures are validated against single-cell imaging data, providing evidence that chromosomal architecture is controlled by a thermodynamics mechanism of polymer phase separation whereby chromatin self-assembles in segregated globules by combinatorial interactions of chromatin factors that include CTCF and cohesin. The thermodynamics degeneracy of single-molecule conformations results in broad structural and temporal variability of TAD-like contact patterns. Globules establish stable environments where specific contacts are highly favored over stochastic encounters. Cohesin depletion reverses phase separation into randomly folded states, erasing average interaction patterns. Overall, globule phase separation appears to be a robust yet reversible mechanism of chromatin organization where stochasticity and specificity coexist.


Q y and Q x Absorption Bands for Bacteriochlorophyll a Molecules from LH2 and LH3.
Light-harvesting systems 2 and 3 (LH2 and LH3) act as antennas for the initial light capture by photosynthetic purple bacteria, thus initiating the conversion of solar energy into chemical energy. The main absorbers are carotenoids and bacteriochlorophylls (BChls), which harvest different parts of the solar spectrum. The first two optical transitions in BChl produce the Q y and Q x absorption bands. The large size of BChl molecules has prevented accurate computational determination of the electronic structures for the relevant states, until we recently succeeded in obtaining the excitation energies and transition dipole moments of the first (Q y) transition for all BChls in LH2 and LH3 using multi-state multiconfigurational second-order perturbation theory calculations. In this work, we go one step further, compute the corresponding values for the Q x transition, in line with previous work [ J. Am. Chem. Soc . 2017 , 139 , 7558 - 7567 ], and compare and assess our data against excitation energies obtained through time-dependent density functional theory methods. Interestingly, we find that the two transitions respond differently to BChls' geometrical factors, such as the macrocycle ring curvature and the dihedral torsion of the acetyl moiety. These findings will aid the unraveling of structure-function relationships for absorption and energy transfer processes in purple bacteria, and once again this demonstrates the viability of multireference quantum chemical methods as computational tools for the photophysics of biomolecules.


Manuscript: Defining Quality Standards for Intensive Home Based Treatment Programs for Youth with Serious Emotional Disorders.
Intensive Home Based Treatment (IHBT) is a critical component of the continuum of community-based behavioral healthcare for youth with serious emotional disorder (SED) and their families. Yet despite being used nationwide at costs of over $100 million annually in some states, a well-vetted, research-based set of quality standards for IHBT has yet to be developed. The current project aimed to define program and practice standards for IHBT, drawing upon literature review, expert interviews, and a systematic Delphi process engaging over 80 participants, including IHBT developers, experts in evidence-based youth mental health, youth and family advocates, IHBT providers, and state policymakers. After two rounds of quantitative and qualitative input, adequate consensus was achieved on 32 IHBT Program Standards and 43 IHBT Practice Standards. These standards hold potential for informing efforts such as development of state regulations, provider contracts, memoranda of agreement, and training and workforce development initiatives. Translation of the quality standards into measurement strategies holds potential for providing a method of continuous quality improvement across multiple levels as well as use in research on IBHT.


Demethylation of induced pluripotent stem cells from type 1 diabetic patients enhances differentiation into functional pancreatic β cells.
Type 1 diabetes (T1D) can be managed by transplanting either the whole pancreas or isolated pancreatic islets. However, cadaveric pancreas is scarcely available for clinical use, limiting this approach. As such, there is a great need to identify alternative sources of clinically usable pancreatic tissues. Here, we used induced pluripotent stem (iPS) cells derived from patients with T1D to generate glucose-responsive, insulin-producing cells (IPCs) via 3D culture. Initially, T1D iPS cells were resistant to differentiation, but transient demethylation treatment significantly enhanced IPC yield. The cells responded to high-glucose stimulation by secreting insulin in vitro The shape, size, and number of their granules, as observed by transmission electron microscopy, were identical to those found in cadaveric β cells. When the IPCs were transplanted into immunodeficient mice that had developed streptozotocin-induced diabetes, they promoted a dramatic decrease in hyperglycemia, causing the mice to become normoglycemic within 28 days. None of the mice died or developed teratomas. Because the cells are derived from "self," immunosuppression is not required, providing a much safer and reliable treatment option for T1D patients. Moreover, these cells can be used for drug screening, thereby accelerating drug discovery. In conclusion, our approach eliminates the need for cadaveric pancreatic tissue.


Prepregnancy Weight in Women with Type I Diabetes Mellitus: Effect on Pregnancy Outcomes.
Objective This study aims to evaluate the association between prepregnancy body mass index (BMI) and adverse pregnancy outcomes in women with type 1 diabetes mellitus (DM). Methods This is a secondary analysis of a cohort of 426 pregnancies in women with type 1 DM recruited before 20 weeks gestation. Women were categorized according to prepregnancy BMI: low BMI (< 20 kg/m2), normal BMI (20 to < 25 kg/m2), and high BMI (≥ 25 kg/m2). The outcomes of interest were: spontaneous abortion (delivery < 20 weeks gestation); preeclampsia; emergent delivery for maternal indications (hypertension or placental abruption); and preterm delivery (< 37 weeks gestation). Analyses included proportional hazards and multiple logistic regression models with covariates: age, age at diagnosis of type 1 DM, previous spontaneous abortion, microvascular disease (nephropathy or retinopathy), and glycohemoglobin A1 concentrations. Results Low BMI was associated with preterm delivery. High BMI was associated with emergent delivery for maternal indications. Glycemic control as measured by glycohemoglobin A1 was associated with increased risk of spontaneous abortion, attenuating the association with low prepregnancy weight. Conclusion Prepregnancy BMI is a risk factor to be considered when caring for women with type 1 DM, in particular for preterm delivery (low BMI) and emergent delivery for maternal indications (high BMI).


Molecular phylogeny of the filaria genus Onchocerca with special emphasis on Afrotropical human and bovine parasites.
Filarial parasites of the genus Onchocerca are found in a broad spectrum of ungulate hosts. One species, O. volvulus, is a human parasite that can cause severe disease (onchocerciasis or 'river blindness'). The phylogenetic relationships and the bionomics of many of the nearly 30 known species remain dubious. Here, the phylogeny of 11 species representing most major lineages of the genus is investigated by analysing DNA sequences from three mitochondrial genes (ND5, 12S and 16S rRNA) and portions of the intergenic spacer of the nuclear 5s rRNA. Special emphasis is given to a clade containing a yet unassigned specimen from Uganda (O. sp. 'Siisa'), which appears to be intermediate between O. volvulus and O. ochengi. While the latter can be differentiated by the O-150 tandem repeat commonly used for molecular diagnostics, O. volvulus and O. sp.'Siisa' cannot be differentiated by this marker. In addition, a worm specimen from an African bushbuck appears to be closely related to the bovine O. dukei and represents the basal taxon of the human/bovine clade. At the base of the genus, our data suggest O. flexuosa (red deer), O. ramachandrini (warthog) and O. armillata (cow) to be the representatives of ancient lineages. The results provide better insight into the evolution and zoogeography of Onchocerca. They also have epidemiological and taxonomic implications by providing a framework for more accurate molecular diagnosis of filarial larvae in vectors.


Intracerebellar hemorrhage due to rupture of angiomatous malformations in the cerebellum.
Two infants were reported with intracerebellar hemorrhage due to rupture of angiomatous malformations in the cerebellum. There have been only a few reports on intracerebellar hemorrhage under 6 years of age, which was due to angiomatous malformation in all cases. The results of operations for intracerebellar hemorrhage in infants were more favorable than those in adults, so even if the infant patient shows severe neurological symptoms preoperatively, an operation should be performed.


Structural studies of Helix aspersa agglutinin complexed with GalNAc: A lectin that serves as a diagnostic tool.
Lectins belong to a differentiated group of proteins known to possess sugar-binding properties. Due to this fact, they are interesting research targets in medical diagnostics. Helix aspersa agglutinin (HAA) is a lectin that recognizes the epitopes containing α-d-N-acetylgalactosamine (GalNAc), which is present at the surface of metastatic cancer cells. Although several reports have already described the use of HAA as a diagnostic tool, this protein was not characterized on the molecular level. Here, we present for the first time the structural information about lectin isolated from mucus of Helix aspersa (garden snail). The amino acid sequence of this agglutinin was determined by Edman degradation and tertiary as well as quaternary structure by X-ray crystallography. The high resolution crystal structure (1.38Å) and MALDI-TOF mass spectrometry analysis provide the detailed information about a large part of the HAA natural glycan chain. The topology of the GalNAc binding cleft and interaction with lectin are very well defined in the structure and fully confirmed by STD HSQC NMR spectroscopy. Together, this provides structural clues regarding HAA specificity and opens possibilities to rational modifications of this important diagnostic tool.


[Teaching clinical decision making in Trondheim].
In the new medical curriculum at the University of Trondheim the preliminary teaching in clinical epidemiology is organized as a series of courses held in the third and fourth year. The objective is to develop skills in clinical appraisal based on scientific knowledge. A workbook and answers to exercises are provided for each course. The workbooks contain short introductions to important clinical epidemiological principles, as well as clinical problems which the students try to solve. The students' activities vary from reading to working with exercises and discussing the various topics. The courses give the students the opportunity to practice their newly acquired knowledge and to apply it to their clinical thinking.


High Selectivity Cofactor NADH Regeneration Organic Iridium Complexes Used for High-Efficiency Chem-Enzyme Cascade Catalytic Hydrogen Transfer.
Our research demonstrated that novel pentamethylcyclopentadienyl (Cp*) iridium pyridine sulfonamide complex PySO2NPh-Ir (7) could highly specifically catalyze nicotinamide adenine dinucleotide (NAD+) into the corresponding reducing cofactor NADH in cell growth media containing various biomolecules. The structures and catalytic mechanism of 7 were studied by single-crystal X-ray, NMR, electrochemical, and kinetic methods, and the formation of iridium hydride species Ir-H was confirmed to be the plausible hydride-transfer intermediate of 7. Moreover, benefiting from its high hydrogen-transfer activity and selectivity for NADH regeneration, 7 was used as an optimal metal catalyst to establish a chem-enzyme cascade catalytic hydrogen-transfer system, which realized the high-efficiency preparation of l-glutamic acid by combining with l-glutamate dehydrogenase (GLDH).


Systemic inflammation worsens outcomes in emergency surgical patients.
Acute care surgeons are uniquely aware of the importance of systemic inflammatory response and its influence on postoperative outcomes; concepts like damage control have evolved from this experience. For surgeons whose practice is mostly elective, the significance of such systemic inflammation may be underappreciated. This study sought to determine the influence of preoperative systemic inflammation on postoperative outcome in patients requiring emergent colon surgery. Emergent colorectal operations were identified in the American College of Surgeons National Surgical Quality Improvement Program 2008 dataset. Four groups were defined by the presence and magnitude of the inflammatory response before operation: no inflammation, systemic inflammatory response syndrome (SIRS), sepsis, or severe sepsis/septic shock. Thirty-day survival was analyzed by Kaplan-Meier method. A total of 3,305 patients were identified. Thirty-day survival was significantly different (p < 0.0001) among the four groups; increasing magnitudes of preoperative inflammation had increasing probability of mortality (p < 0.0001). Hazard ratios indicated that, compared with patients without preoperative systemic inflammation, the relative risk of death from SIRS was 1.9 (p < 0.0001), from sepsis was 2.5 (p < 0.0001), and from severe sepsis/septic shock was 6.7 (p < 0.0001). Operative time of <150 minutes was associated with decreased risk of morbidity (odds ratio = 0.64; p < 0.0001). Upregulation of the systemic inflammatory response is the primary contributor to death in emergency surgical patients. In SIRS or sepsis patients, operations <2.5 hours are associated with fewer postoperative complications. These results further reinforce the concept of timely surgical intervention and suggest a potential role for damage control operations in emergency general surgery. II, prognostic study.


Quantification of allergenic bovine milk α(S1)-casein in baked goods using an intact ¹⁵N-labeled protein internal standard.
Intact bovine ¹⁵N-α(S1)-casein was used as an internal standard in a selected reaction monitoring (SRM) assay for milk protein in baked food samples containing fats, sugar, and gums. Effects on SRM results of sample matrix composition in two biscuit recipes containing nonfat dry milk (NFDM) were studied, including samples from a milk allergen ELISA proficiency trial. Following extraction of defatted samples with carbohydrate-degrading enzymes and acid precipitation of casein, the SRM assay exhibited an LOQ of <3 ppm NFDM with 60-80% recovery. NFDM levels measured by the SRM assay were 1.7-2.5 times greater than median levels determined by ELISA. Differences were observed in the α(S1)-casein interpeptide SRM ion abundance profile between recipes and after baking. ¹⁵N-α(S1)-Casein increases SRM analysis accuracy by correcting for extraction recovery but does not eliminate underestimation of allergen concentrations due to baking-related milk protein transformation (modifications).


Persistence of subclinical deformed wing virus infections in honeybees following Varroa mite removal and a bee population turnover.
Deformed wing virus (DWV) is a lethal virus of honeybees (Apis mellifera) implicated in elevated colony mortality rates worldwide and facilitated through vector transmission by the ectoparasitic mite Varroa destructor. Clinical, symptomatic DWV infections are almost exclusively associated with high virus titres during pupal development, usually acquired through feeding by Varroa mites when reproducing on bee pupae. Control of the mite population, generally through acaricide treatment, is essential for breaking the DWV epidemic and minimizing colony losses. In this study, we evaluated the effectiveness of remedial mite control on clearing DWV from a colony. DWV titres in adult bees and pupae were monitored at 2 week intervals through summer and autumn in acaricide-treated and untreated colonies. The DWV titres in Apistan treated colonies was reduced 1000-fold relative to untreated colonies, which coincided with both the removal of mites and also a turnover of the bee population in the colony. This adult bee population turnover is probably more critical than previously realized for effective clearing of DWV infections. After this initial reduction, subclinical DWV titres persisted and even increased again gradually during autumn, demonstrating that alternative non-Varroa transmission routes can maintain the DWV titres at significant subclinical levels even after mite removal. The implications of these results for practical recommendations to mitigate deleterious subclinical DWV infections and improving honeybee health management are discussed.


Tomato spotted wilt virus Can Infect Resistant Tomato when Western Flower Thrips Inoculate Blossoms.
Tomato spotted wilt is a major disease of crops worldwide. Resistant cultivars carrying the Sw-5 allele for resistance to tomato spotted wilt disease (TSW) provide the most effective control method in tomato (Solanum lycopersicum). However, infections of fruit on Sw-5+ tomato plants suggest the virus resistance may not be fully expressed in blossoms or developing fruit. The objective of this study was to determine if the thrips vector, the western flower thrips (Frankliniella occidentalis), can transmit non-resistance breaking Tomato spotted wilt virus (TSWV) isolates when confined to blossoms on plants with and without the Sw-5 resistance allele. Twenty-one percent of 33 Sw-5+ plants inoculated by adult thrips feeding on blossom clusters or small fruit developed infections in the reproductive tissue, whereas 68% of 25 Sw-5- plants developed infections. Systemic infections also occurred following inoculation of blossoms in host genotypes with and without Sw-5. These results were further supported by field experiments that showed high proportions of infected fruit as well as a limited infection of foliage on the same stem as the infected fruit in Sw-5+ plants when F. occidentalis were abundant in blossoms. These findings help to explain observations of abundant late season infections of Sw-5 cultivars in commercial plantings and suggest that management of F. occidentalis infestations during the bloom period may be important for effective management of TSWV in susceptible tomato cultivars as well as cultivars expressing the Sw-5 allele for TSW resistance.


Amino acids levels as a potential biomarker in myasthenia gravis.
The search for new diagnostic and therapeutic approaches for myasthenia gravis (MG) is highly desirable. Therefore, the aim of the present study is to assess the profile of non-essential amino acids in the serum of MG patients. We evaluated the serum levels of non-essential amino acids in MG patients (n = 10) and control subjects (CS, n = 10) using high-performance liquid chromatography (HPLC) method and assuming a two-fold concentration difference between the groups as significant. Serum levels of aspartic acid and glutamic acid in MG patients were significantly higher than in CS. There were no significant differences in mean serum levels of glycine, proline, alanine, serine, cysteine, arginine and tyrosine between MG patients and CS. The results indicate the need for further research to assess the role of non-essential amino acids in MG. Moreover, our preliminary results suggest that the metabolism of some amino acids seems to be changed in patients with MG. Therefore, we conclude that amino acids profiling might be helpful in searching for putative biomarkers of the central nervous system diseases such as myasthenia gravis.


A randomized, double-blind, placebo-controlled trial of onabotulinumtoxin A trigger point injections for myofascial pelvic pain.
Pelvic pain is estimated to effect 15% of women, and onabotulinumtoxin A is used to treat a variety of pain disorders. However, the data on the use of onabotulinumtoxin A for the treatment of women with myofascial pelvic pain are limited. The objective of the study was to compare the effect of onabotulinumtoxin A vs placebo injections to the pelvic floor muscles in women with myofascial pelvic pain. This was a double-blind, randomized, placebo-controlled trial in women with myofascial pelvic pain. Women ≥18 years were eligible if they reported pain ≥6 on a 10 point visual analog scale ≥50% of the time and had pain on palpation ≥6 on the visual analog scale in ≥1 of 6 pelvic floor muscle groups. Participants were randomly allocated to a pelvic floor injection of 200 units of onabotulinumtoxin A or 20 mL of saline. All participants started 8 weeks of physical therapy 4 weeks after the injection. Participants completed validated questionnaires at baseline, 2, 4, and 12 weeks after injection. At each visit, a urogynecologist who was blinded to treatment arm performed a clinical examination with palpation of the left and right sides of 6 pelvic floor muscle groups. The primary outcome was change in participant-reported pain on palpation of the most painful pelvic floor muscle at 2 weeks. Analyses were intention to treat. We consented 60 women. One participant was lost to follow-up after she was consented; therefore, we randomized 59 women. The groups had similar demographic and clinical characteristics. With regard to the primary outcome, there was no significant difference between the intervention and placebo groups in the change in participant-reported pain on palpation of the most painful pelvic floor muscle at 2 weeks. There were no significant differences in participant-reported pain on palpation for any muscle group at 4 or 12 weeks. At 4 and 12 weeks, participants in the intervention group reported greater declines in overall pelvic pain on the visual analog scale compared with the placebo group, although these differences were not statistically significant (both P = .16). Using the Patient Global Impression of Improvement index, participants in the intervention group were more likely to report their symptoms were improved at 4 and 12 weeks compared with the placebo group, although this difference was significant only at 4 weeks (P = .03 and P = .10, respectively). At 2 weeks, the placebo group had a significant improvement in the Pelvic Floor Distress Inventory score compared with the intervention group (P = .01); however, this difference did not persist at 4 (P = .19) or 12 weeks (P = .11). At 2 weeks, the most common adverse event was constipation in the intervention and placebo groups, with 10.1% reporting de novo constipation. This was followed by urinary incontinence in the intervention group (22%) and urinary tract infection (9%) in the placebo group. Pelvic floor onabotulinumtoxin A injections for myofascial pelvic pain were not more effective than saline injections at decreasing muscle pain on palpation. Despite this, participants who received onabotulinumtoxin A were more likely than those who received saline to report improvement, albeit not statistically significant, in their overall pelvic floor pain at 4 and 12 weeks.


On the mechanism of lactate dehydrogenase leakage from normal and D-galactosamine-treated hepatocytes in monolayer culture.
Synthesis, degradation and leakage of lactate dehydrogenase and of total protein was measured using D-galactosamine-treated rat hepatocytes in monolayer culture. The kinetics of [3H]leucine incorporation into trichloroacetic acid-precipitable material and into isolated lactate dehydrogenase of cells and of the extra-cellular space revealed a similar extent of inhibition of both synthesis and leakage following exposure to D-galactosamine. Hepatocyte cultures that had been labeled before D-galactosamine treatment lost intracellular protein-associated radioactivity almost as rapidly as control cells up to the time of measurable enzyme leakage; thereafter, the rate of 3H-loss increased in the treated cells. Lactate dehydrogenase present in the medium is degraded less rapidly than the enzyme in the intracellular space. This explains the apparent increase of total lactate dehydrogenase activity in D-galactosamine-treated as compared to control cultures. Following [3H]leucine addition to D-galactosamine-treated cultures, the specific radioactivity of the leaked lactate dehydrogenase in the medium was never greater than that of the enzyme in the cytosolic compartment. The data rule out a direct excretion of newly synthesized enzyme as a result of D-galactosamine action.


Electron microscopic and molecular identification of Wolbachia endosymbionts from Onchocerca lupi: implications for therapy.
It was recently demonstrated that Wolbachia intracellular bacteria (alpha 2 proteobacteria, Rickettsiales) living in filarial nematodes are obligatory symbionts of their hosts. Herein, we report the electron microscopic and 16S ribosomal DNA-based (16S rDNA) identification of the endobacteria harboring in Onchocerca lupi. The worm nodules containing the nematodes were removed from three Hungarian dogs naturally infected with O. lupi. Wolbachia-like endobacteria were detected by electron microscopy in the lateral chords of both adult worms and microfilariae. The endosymbionts in O. lupi resemble in location, size, and morphology the wolbachiae found in other filariae. The presence of wolbachiae in O. lupi was also confirmed by PCR amplification of the 16S rDNA of the bacteria. The 16S rDNA-based phylogenetic analysis revealed that the endosymbionts of O. lupi infecting dogs belong to the supergroup C of Wolbachia pipientis and are not identical with those of other Onchocerca spp. sequenced so far. Since intermittent treatment with oxytetracycline has adulticid and microfilaricid activity by depletion of Wolbachia endobacteria, this antibiotic treatment regimen may offer an alternative of ivermectin or diethylcarbamazine in the suppression of postoperative microfilaridermia in Onchocerca-infected dogs and may prevent relapse.


Partial properdin deficiency.
The action of properdin in supporting complement consumption has been investigated in the serum from two individuals with partial properdin deficiency. Our studies are in agreement with those in which serum were artificially depleted of properdin in showing that serum deficient in properdin results in less C3 consumption in the presence of activators of both the alternative and classical complement pathways. Properdin deficiency did not lessen C3 consumption by C3NeF or CoF, confirming that properdin is not necessary for the action of these substances. Our findings indicate that partial properdin deficiency is innocuous but that complete absence of properdin may severely limit one's ability amplify complement activation.


U.S. masters track participation reveals a stable sex difference in competitiveness.
It is well established that men are more likely than women to engage in direct competition, but it is unclear if this reflects social structural conditions or evolved predispositions. These hypotheses can be addressed by quantifying competitiveness in sports and testing if the sex difference is decreasing over time in the U.S., a society where social roles have converged. Study 1 assessed participation and the occurrence of relatively fast performances by masters runners (40-74 years old) at recent road races and track meets. Fast performances occurred over 20 times more often at track meets than at road races. Women comprised 55% of finishers at roads races but only 15-28% of finishers at track meets. Thus, the sex difference in masters track participation can serve as a measure of the sex difference in competitiveness. Study 2 used data from national championship meets and yearly rankings lists to test whether the sex difference in masters track participation decreased from 1988-2012. The sex difference decreased overall, but there was no evidence of change since the late 1990s. Therefore, the sex difference in the willingness to engage in direct sports competition appears to reflect both social structural conditions and evolved predispositions.


Essential elements in determining patient's complexity of nursing care.
Nurses dedicate majority of working time to satisfy patients' needs, which are oftentimes complex. Although the concept of patient's complexity of care (PCC) has been extensively studied, it remains undefined in its essential characteristics. Various tools for assessing PCC have been developed, yet, there is no gold standard of reference. This study intends to explore the elements that determine PCC focusing on the patient's needs. The bed management team of a University Hospital developed in 2010 a PCC measurement instrument, similar to a triage system, to classify rapidly patients into macro-levels of care complexity (low, medium, high, intensive). Descriptive analysis of prospectively collected data registered in nursing records during 2016-2017 are presented in this study. The incidence of individual metrics on the value assigned to the overall PCC level was calculated. 26593 patients' records were analyzed. The final level of PCC was equal to patient's level of autonomy in 92,2% of cases, to clinical stability in 74,4% and to cognitive status in 58,4%. Our finding indicate that essential elements to determine PCC are patient's autonomy and clinical stability. Therefore, nurses can use these metrics to classify quickly PCC in their work settings. Identifying and measuring PCC is very important for nurses. It can facilitate and support nurse decision-making in design, implementation and evaluation of activities aimed at improving patient care. Moreover, it can help in the distribution of care loads in the shift, and from an organizational point of view, it can allow defining staffing assets.


Genome-editing technologies and patent landscape overview.
Unlike with zinc finger nuclease and transcriptional activator-like effector nuclease DNA modification technologies that rely on lead proteins, developed through expensive and time-consuming processes, the clustered regularly interspaced short palindromic repeat (CRISPR)/Cas system has rapidly emerged as the most promising gene-editing technology to date for the modification of any selected DNA sequence. CRISPR is receiving tremendous fanfare due, in part, to its potential to provide a means to fundamentally alter medical genetics and especially cancer medicine. In this review, we compare key technologies of genome-editing zinc finger nucleases, transcriptional activator-like effector nucleases and CRISPR, with a focus on the race to acquire lucrative intellectual property rights, the current CRISPR patent dispute and potential repercussions on innovation and the adoption of this promising technology by the medical community.


Terahertz spectra and normal mode analysis of the crystalline VA class dipeptide nanotubes.
Terahertz (THz) vibrational modes are characterized by nonlocal, collective molecular motions which are relevant to conformational changes and molecular functions in biological systems. We have investigated the THz spectra of a set of small bionanotubes which can serve as very simple models of membrane pores, and have examined the character of the THz modes which can impact transport processes. In this work, THz spectra of the crystalline VA class dipeptide nanotubes were calculated at both the harmonic and vibrational self-consistent field (VSCF) level using the CHARMM22 force field with periodic boundary conditions. Comparison of the calculated THz spectra against the experimental spectra revealed that the VSCF corrections generally improved the predictions in the low-frequency region. The improvements were especially manifested in the overall blue-shifts of the VSCF frequencies relative to the harmonic values, and blue shifts were attributed to the overall positive coupling strengths in all systems. Closer examination of the motions in the most significantly coupled normal mode pairs leads us to propose that, when two similar side-chain squeezing modes are coupled, the rapidly increased van der Waals interactions can lead to a stiffening of the effective potential, which in turn leads to the observed blue-shifts. However, we also noted that when the side-chain atoms become unphysically proximate and the van der Waals repulsion becomes too large, the VSCF calculations tend to deviate in the high frequency region and for the system of l-isoleucyl-l-valine. In addition, normal-mode analysis revealed a series of channel-breathing motions in all systems except l-valyl-l-alanine. We show that the inner products of the backbone vibrations between these channel-breathing motions divided the remaining VA class dipeptide systems into two subgroups. It is suggested that these modes may facilitate a pathway for the guest molecule absorption, substitution and removal in the VA class dipeptide nanotubes. Normal mode analysis also demonstrated that the THz motions may contribute to the pore permeability either directly by changing the pore size, or indirectly by affecting the solvent-host effective potentials.


Towards evidence-based emergency medicine: best BETs from the Manchester Royal Infirmary. BET 2: Therapeutic hypothermia for cardiac arrest of asphyxial origin.
A short-cut review was carried out to establish whether therapeutic hypothermia leads to improved neurological outcomes in adult patients who have sustained a cardiac arrest of asphyxial origin. Four studies were directly relevant to the question. The author, date and country of publication, patient group studied, study type, relevant outcomes, results and study weaknesses of these papers are shown in table 2. The clinical bottom line is that evidence for the use of therapeutic hypothermia in patients who are comatose following asphyxia is very limited, which precludes an evidence-based recommendation. A large, multicentre randomised controlled trial is necessary to answer this question. Our review has therefore highlighted an important area for further research.


mGen--An Open Source Framework for Generating Clinical Documents.
As formalised electronic storage of medical data becomes more and more wide spread the possibility and need for creating human readable presentations for various purposes increases. This paper presents the mGen framework - a general framework for text generation, written in Java and in the process of being open-sourced, that has been developed within the MedView projectThe work presented in this paper was supported by the Swedish Agency for Innovation Systems (VINNOVA).. The framework has been used for several years to generate literally thousands of clinical documents at an oral medicine clinic in Sweden.


Reinventing postgraduate training in the plant sciences: T-training defined through modularity, customization, and distributed mentorship.
The Plant Science Research Network (PSRN) comprises scientific societies and organizations with a mission to build and communicate a consensus vision of the future of plant science research, education, and training. This report enumerates a set of far-reaching recommendations for postgraduate training that emerged from workshops held in October 2016 and September 2017. These recommendations broaden and deepen the T-training concept presented in the Decadal Vision for Plant Science, which emphasizes experiential learning beyond the traditional disciplinary focus. Both workshops used the scenarios developed in Imagining Science in 2035 as a mechanism to encourage out-of-the-box thinking, an approach that led to the innovative recommendations and solutions described here. At the heart of our recommendations is the empowerment of trainees, who should be enabled to customize and take ownership of their training experiences. This fundamental concept is embodied in five principles: (a) Trainees should be provided guidance and resources needed to define and pursue career objectives within and beyond academia, conferring to them greater independence and responsibility in shaping their own future. (b) Learning should be flexible, adaptable, and distributed. Training should combine traditional and modular coursework to encompass both technical and professional skills. Guidance from diverse mentoring teams will support and tailor training toward diverse, personalized career paths. (c) Scientific research experiences should be broad and question-driven, whether motivated by basic discovery or seeking solutions to societal challenges. Trainees should continue to gain mastery of one or a few core scientific disciplines and their key tools and approaches. (d) Trainees should be skilled in science communication and incentivized to engage with and learn from the broader public community, helping to maintain an active dialogue among public, private, and academic sectors. (e) Training programs should foster and facilitate the inclusion of individuals with a diverse range of life experiences and should prioritize trainee well-being. The report recommendations call for a profound cultural shift, one that embraces and extends educational delivery trends toward self-learning and distance learning, considers trainee well-being as an essential requirement for success, and acknowledges the importance of effective two-way communication with the public. This shift is intended to broaden participation in the plant science workforce, both in terms of diversity and numbers, while maintaining excellence in core scientific training. Cultural change takes time, but among academic institutions the need for significant change and innovation in postgraduate training is increasingly pressing. As such, the immediate intent is for these recommendations to catalyze pilot programs and also build on emergent prototypes that exist globally while creating momentum for larger scale changes over longer time periods.


Prognostic correlation of HLA-DR expression in large cell lymphoma as determined by LN3 antibody staining. An Eastern Cooperative Oncology Group (ECOG) study.
Several previous studies employing surface-marker techniques suggest that HLA-DR antigen expression may correlate with prognosis in diffuse large cell lymphoma. Control studies in the authors' laboratory indicated that LN3 positivity in paraffin-embedded tissue correlates well with HLA-DR antigen expression. Accordingly they examined 212 cases of diffuse large cell lymphoma from one of Eastern Cooperative Oncology Group's (ECOG) high-grade malignant lymphoma treatment protocols. All patients studied had careful clinical follow-up of between 5 and 11 1/2 years from diagnosis and initiation of therapy. There was no correlation of strongly positive, positive, weakly positive, equivocal, or negative staining with survival. Cases that were LN3 negative had a median survival of 18.8 months versus a median survival of 39.4 months for all LN3-positive cases. However this difference was not statistically significant when using a log-rank test. The authors were unable to confirm that HLA-DR antigen expression, as defined by LN3, is prognostically valuable in diffuse large cell lymphoma in a multi-institutional study.


Effect of indomethacin on electrical field stimulation-induced contractions of isolated transverse and longitudinal rat gastric fundus strips.
To study the effects of indomethacin on the isolated transverse and longitudinal rat gastric fundus strips. The strips were suspended in an organ bath containing oxygenated Krebs solution, and contractile responses to electrical field stimulation were recorded on a physiograph in an isotonic manner after administration of cumulative concentrations of indomethacin. The effects of indomethacin on the strips pretreated with K(ATP) channel modulators, diazoxide and glybenclamide were studied. Treatment of the transverse strips with indomethacin resulted in a concentration-dependent inhibitory response. In longitudinal strips, biphasic responses were seen, which included a stimulatory response at low concentrations of indomethacin, followed by an inhibitory response at higher concentrations. Diazoxide pre-treatment inhibited the stimulatory response of longitudinal strips. Glybenclamide pre-treatment not only blocked inhibitory effect of the low concentrations of indomethacin on transverse strips, but also increased the amplitude of contractions. Moreover, the drug decreased the amplitude of contractions in longitudinal strips. Responses of the isolated longitudinal and transverse rat gastric fundus strips to indomethacin are not similar, and are influenced by K(ATP) channel modulators.


Fixation of length-stable femoral shaft fractures in heavier children: flexible nails vs rigid locked nails.
The treatment of femoral shaft fractures in heavier children has been studied extensively, however, no study has directly compared elastic nail (TEN) and rigid locking nails (RLN) in this population. Our goal was to compare TEN with RLN in length-stable diaphyseal femur fractures in heavier children and adolescents (47 to 85 kg) using weight-matched cohorts. We retrospectively reviewed records for length-stable diaphyseal femoral fractures treated with TEN or RLN over 8 years at our Level 1 Pediatric Trauma Center. Perioperative and follow-up data, including total charges for care, were recorded and radiographic images were reviewed. These data were used to create 2 cohorts having each patient in the TEN cohort matched to within 2 kg of the corresponding patient in the RLN cohort. Fifteen patients from each cohort could be weight matched (TEN, 60.8 kg vs. RLN, 60.4 kg). The RNL cohort was older (15.4 vs. 13.5 y; P = 0.005). Time in operating room and estimated blood loss were greater in the RLN cohort: 158 versus 220 minutes (P = 0.003) and 42 versus 182 ml (P = 0.003), respectively. All patients had a full range of motion at the latest follow-up. Complications were observed in 6 of 15 TEN and 10 of 15 RNL (P = 0.14). Implant-related problems were more common in RLN patients, but this was not statistically significant (3 of 15 vs. 9 of 15; P = 0.06). In the TEN cohort, malunion and leg length discrepancy (> 2 cm) each occurred in a single patient (20 degrees varus, 2.3 cm shortening, respectively) compared with 0 of 15 in RLN (P = 0.48). Treatment with TEN resulted in a total charge of $742 more than RLN (P = 0.75). In our weight-matched comparison, the use of TEN resulted in decreased time in operating room, estimated blood loss, and implant-related problems. Malunion and leg length discrepancy remain of concern when heavier patients are treated by TEN, but were not significantly increased relative to RNL in this study.


Lutropin-dependent protein phosphorylation and steroidogenesis in rat tumour Leydig cells.
Tumour Leydig cells have been incubated in the presence or absence of lutropin (luteinizing hormone, ;LH'). Stimulation of cells with lutropin (1000ng/ml) in the presence of 1-methyl-3-isobutylxanthine (0.25mm) resulted in increased steroid production and increased protein phosphorylation. When pregnenolone metabolism was inhibited, basal pregnenolone production was 26.9+/-7.4ng/60min per 10(6) cells; stimulated production was 156.1+/-39.5ng/60min per 10(6) cells (means+/-s.d., n=4). Lutropin-dependent phosphorylated proteins of molecular mass 17000, 22000, 24000, 33000 and 57000Da were detected. A significant increase of [(32)P]P(i) incorporation into these phosphorylated proteins was observed concomitant with the increased pregnenolone production. The occurrence of the phosphoproteins in nuclei, mitochondria and postmitochondrial-supernatant was investigated. The 17000Da phosphoprotein was found in the nuclear fraction, whereas the 22000, 24000, 33000 and 57000Da phosphoproteins were localized in the postmitochondrial-supernatant fraction. Of the cholesterol-side-chain-cleavage activity, 80.3+/-6.1% (mean+/-s.d., n=5) was present in the mitochondrial fraction isolated from tumour Leydig cells, and this activity was 2.5-fold increased when cells had been preincubated with lutropin/1-methyl-3-isobutylxanthine (basal production: 194.6+/-28.6ng/30min per mg of protein; lutropinstimulated production: 498.8+/-91.5ng/30min per mg of protein; means+/-s.d., n=3). The similarities in the kinetics of the phosphorylation of proteins and the pregnenolone production after addition of lutropin/1-methyl-3-isobutylxanthine indicate that the phosphoproteins could be involved in the lutropin-dependent increase in steroidogenesis in tumour Leydig cells. It remains to be demonstrated, however, to what extent the phosphoproteins outside the mitochondria can influence the cholesterol-side-chain-cleavage activity inside the mitochondria.


Loneliness and the effects of life changes.
In this study, factor analysis was used to examine whether men and women experience loneliness differently and how marital status affects the experience of loneliness (especially if the marriage is terminated). The experiences of chronic and situational or event-related loneliness are also compared. Finally, the study is also an investigation of the social stigma that causes people to refuse to admit their loneliness. As a result of this stigma, when questioned for research purposes, individuals usually report a recall past loneliness rather than a current or ongoing experience. In the present study, the difference between loneliness recalled and loneliness experienced at the time of questioning is investigated.


Wound healing activity of Carica papaya L. in experimentally induced diabetic rats.
The aqueous extract of C. papaya fruit (100 mg kg(-1) day(-1) for 10 days) was evaluated for its wound healing activity in streptozotocin-induced diabetic rats using excision and dead space wound models. Extract-treated animals exhibited 77% reduction in the wound area when compared to controls which was 59%. The extract treated wounds were found to epithelize faster as compared to controls. The wet and dry granulation tissue weight and hydroxyproline content increased significantly when compared to controls. The extract exhibited antimicrobial activity against the five organisms tested. Carica papaya promotes significant wound healing in diabetic rats and further evaluation of this activity in humans is suggested.


Chemotherapy and radiation therapy elicits tumor specific T cell responses in a breast cancer patient.
Experimental evidence and clinical studies in breast cancer suggest that some anti-tumor therapy regimens generate stimulation of the immune system that accounts for tumor clinical responses, however, demonstration of the immunostimulatory power of these therapies on cancer patients continues to be a formidable challenge. Here we present experimental evidence from a breast cancer patient with complete clinical response after 7 years, associated with responsiveness of tumor specific T cells. T cells were obtained before and after anti-tumor therapy from peripheral blood of a 63-years old woman diagnosed with ductal breast cancer (HER2/neu+++, ER-, PR-, HLA-A*02:01) treated with surgery, followed by paclitaxel, trastuzumab (suspended due to cardiac toxicity), and radiotherapy. We obtained a leukapheresis before surgery and after 8 months of treatment. Using in vitro cell cultures stimulated with autologous monocyte-derived dendritic cells (DCs) that produce high levels of IL-12, we characterize by flow cytometry the phenotype of tumor associated antigens (TAAs) HER2/neu and NY-ESO 1 specific T cells. The ex vivo analysis of the TCR-Vβ repertoire of TAA specific T cells in blood and Tumor Infiltrating Lymphocytes (TILs) were performed in order to correlate both repertoires prior and after therapy. We evidence a functional recovery of T cell responsiveness to polyclonal stimuli and expansion of TAAs specific CD8+ T cells using peptide pulsed DCs, with an increase of CTLA-4 and memory effector phenotype after anti-tumor therapy. The ex vivo analysis of the TCR-Vβ repertoire of TAA specific T cells in blood and TILs showed that whereas the TCR-Vβ04-02 clonotype is highly expressed in TILs the HER2/neu specific T cells are expressed mainly in blood after therapy, suggesting that this particular TCR was selectively enriched in blood after anti-tumor therapy. Our results show the benefits of anti-tumor therapy in a breast cancer patient with clinical complete response in two ways, by restoring the responsiveness of T cells by increasing the frequency and activation in peripheral blood of tumor specific T cells present in the tumor before therapy.


Synonymous rare arginine codons and tRNA abundance affect protein production and quality of TEV protease variant.
It has been identified that a TEV protease (TEVp) variant, TEVp(5M), displays improved solubility. Here, we constructed fifteen TEVp(5M) variants with one or more of six rare arginine codons in the coding sequence replaced with abundant E. coli arginine codons. These codon variants expressed in either E. coli BL21 (DE3) or Rossetta (DE3) showed different solubility and activity. Supply of rare tRNAs increased the tendency of certain codon variants to form insoluble aggregates at early induction stage, as determined by the fused S-tag. About 32% increase in soluble protein production of M5 variant with four synonymously mutated arginine codons was identified in Rossetta (DE3) cells using GFP fusion reporter, comparable to that of TEVp(5M). After purification, two other codon variants from both E. coli strains exhibited less activity than TEVp(5M) on cleaving the native or modified recognition sequence incorporated between GST and E. coli diaminopropionate ammonialyase by enzyme-coupled assay, whereas purified M5 variant showed activity similar to the TEVp(5M). Supply of rare tRNAs caused the decrease of activity of TEVp(5M) and M5 by about 21%. Our results revealed that engineering of highly soluble TEVp variants can be achieved by the combined mutations of amino acid residues and optimization of specific rare codons, whereas simple augment of rare tRNAs abundance resulted in partial loss of activity.


Extended duration rivaroxaban versus short-term enoxaparin for the prevention of venous thromboembolism after total hip arthroplasty: a double-blind, randomised controlled trial.
The risk of venous thromboembolism is high after total hip arthroplasty and could persist after hospital discharge. Our aim was to compare the use of rivaroxaban for extended thromboprophylaxis with short-term thromboprophylaxis with enoxaparin. 2509 patients scheduled to undergo elective total hip arthroplasty were randomly assigned, stratified according to centre, with a computer-generated randomisation code, to receive oral rivaroxaban 10 mg once daily for 31-39 days (with placebo injection for 10-14 days; n=1252), or enoxaparin 40 mg once daily subcutaneously for 10-14 days (with placebo tablet for 31-39 days; n=1257). The primary efficacy outcome was the composite of deep-vein thrombosis (symptomatic or asymptomatic detected by mandatory, bilateral venography), non-fatal pulmonary embolism, and all-cause mortality up to day 30-42. Analyses were done in the modified intention-to-treat population, which consisted of all patients who had received at least one dose of study medication, had undergone planned surgery, and had adequate assessment of thromboembolism. This study is registered at ClinicalTrials.gov, number NCT00332020. The modified intention-to-treat population for the analysis of the primary efficacy outcome consisted of 864 patients in the rivaroxaban group and 869 in the enoxaparin group. The primary outcome occurred in 17 (2.0%) patients in the rivaroxaban group, compared with 81 (9.3%) in the enoxaparin group (absolute risk reduction 7.3%, 95% CI 5.2-9.4; p<0.0001). The incidence of any on-treatment bleeding was much the same in both groups (81 [6.6%] events in 1228 patients in the rivaroxaban safety population vs 68 [5.5%] of 1229 patients in the enoxaparin safety population; p=0.25). Extended thromboprophylaxis with rivaroxaban was significantly more effective than short-term enoxaparin plus placebo for the prevention of venous thromboembolism, including symptomatic events, in patients undergoing total hip arthroplasty.


[CGS20267 (Letrozole), a new aromatase inhibitor: late phase II study in postmenopausal women with advanced or recurrent breast cancer (no. 2)--evaluation of efficacy and safety at the recommended clinical dose CGS20267 Study Group].
In the first part of this late phase II study, we determined the recommended clinical dose of CGS20267 to be 1.0 mg once daily for the treatment of postmenopausal women with advanced or recurrent breast cancer. To further evaluate the efficacy and safety of CGS20267 at the derived or recommended clinical dose, 30 more patients were enrolled in the second part of the study, and were added to the patients treated at 1.0 mg in the first part. As a result of putting the first and second parts together, the objective response rate at 1.0 mg was found to be 38.3%, which was almost equal to that of the early phase II study (40.7%). Drug-related adverse events occurred in 35.4% of the patients at 1.0 mg, and all of the events were of grade 2 or lower. These results demonstrated that CGS20267 1.0 mg once daily is effective and well tolerated in the treatment of postmenopausal women with advanced or recurrent breast cancer.


Posttranscriptional silencing of the lncRNA MALAT1 by miR-217 inhibits the epithelial-mesenchymal transition via enhancer of zeste homolog 2 in the malignant transformation of HBE cells induced by cigarette smoke extract.
Lung cancer is regarded as the leading cause of cancer-related deaths, and cigarette smoking is one of the strongest risk factors for the development of lung cancer. However, the mechanisms for cigarette smoke-induced lung carcinogenesis remain unclear. The present study investigated the effects of an miRNA (miR-217) on levels of an lncRNA (MALAT1) and examined the role of these factors in the epithelial-mesenchymal transition (EMT) induced by cigarette smoke extract (CSE) in human bronchial epithelial (HBE) cells. In these cells, CSE caused decreases of miR-217 levels and increases in lncRNA MALAT1 levels. Over-expression of miR-217 with a mimic attenuated the CSE-induced increase of MALAT1 levels, and reduction of miR-217 levels by an inhibitor enhanced expression of MALAT1. Moreover, the CSE-induced increase of MALAT1 expression was blocked by an miR-217 mimic, indicating that miR-217 negatively regulates MALAT1 expression. Knockdown of MALAT1 reversed CSE-induced increases of EZH2 (enhancer of zeste homolog 2) and H3K27me3 levels. In addition to the alteration from epithelial to spindle-like mesenchymal morphology, chronic exposure of HBE cells to CSE increased the levels of EZH2, H3K27me3, vimentin, and N-cadherin and decreased E-cadherin levels, effects that were reversed by MALAT1 siRNA or EZH2 siRNA. The results indicate that miR-217 regulation of EZH2/H3K27me3 via MALAT1 is involved in CSE-induced EMT and malignant transformation of HBE cells. The posttranscriptional silencing of MALAT1 by miR-217 provides a link, through EZH2, between ncRNAs and the EMT and establishes a mechanism for CSE-induced lung carcinogenesis.


[Clinicopathologic study of 9 patients with neuroendocrine carcinoma of the larynx].
To evaluate the clinic manifestation, therapy and prognosis of neuroendocrine carcinoma of the larynx. Nine cases with neuroendocrine carcinoma of the larynx treated between May 2005 and June 2011 were analyzed retrospectively. There were six males and three females, with a median age of 58 years (ranging from 35 to 65 years). Five cases were treated by only operation, and four cases by combined treatment (surgery followed by radiotherapy and chemotherapy). Two patients with typical carcinoid tumor had not any recurrence with following up of 28 and 30 months, respectively. Of three patients with atypical carcinoid tumor, one patient recurred in 36 months after the first operation and followed by re-operation, with no recurrence by further 30 month follow-up, and the other two patients did not recur 15 and 20 month follow-up, respectively. Of three patients with small cell neuroendocrine carcinoma, two died after 11 and 14 months, respectively, and another patient was followed up for 18 months, with no recurrence. One patient with paraganglioma showed no recurrence with a follow up of 32 months. Neuroendocrine carcinoma in larynx included typical carcinoid tumor, atypical carcinoid tumor, small cell neuroendocrine carcinoma and paraganglioma. Accurate diagnosis relies on histopathologic and immunohistochemical examination. There is no standard treatment plan and based-surgery combined treatment should be adopted to laryngeal neuroendocrine carcinoma. The prognosis is dependent on tumor types.


Emerging anticoagulants.
Warfarin, heparin and their derivatives have been the traditional anticoagulants used for prophylaxis and treatment of venous thromboembolism. While the modern clinician is familiar with the efficacy and pharmacokinetics of these agents, their adverse effects have provided the impetus for the development of newer anticoagulants with improved safety, ease of administration, more predictable pharmacodynamics and comparable efficacy. Research into haemostasis and the coagulation cascade has made the development of these newer anticoagulants possible. These drugs include the factor Xa inhibitors and IIa (thrombin) inhibitors. Direct and indirect factor Xa inhibitors are being developed with a relative rapid onset of action and stable pharmacokinetic profiles negating the need for close monitoring; this potentially makes them a more attractive option than heparin or warfarin. Examples of direct factor Xa inhibitors include apixaban, rivaroxaban, otamixaban, betrixaban and edoxaban. Examples of indirect factor Xa inhibitors include fondaparinux, idraparinux and idrabiotaparinux. Direct thrombin inhibitors (factor IIa inhibitors) were developed with the limitations of standard heparin and warfarin in mind. Examples include recombinant hirudin (lepirudin), bivalirudin, ximelagatran, argatroban, and dabigatran etexilate. This review will discuss emerging novel anticoagulants and their use for the prophylaxis and management of venous thromboembolism, for stroke prevention in nonvalvular atrial fibrillation and for coronary artery disease.


CE and nanomaterials - Part II: Nanomaterials in CE.
The scope of this two-part review is to summarize publications dealing with CE and nanomaterials together. This topic can be viewed from two broad perspectives, and this article is trying to highlight these two approaches: (i) CE of nanomaterials, and (ii) nanomaterials in CE. The second part aims at summarization of publications dealing with application of nanomaterials for enhancement of CE performance either in terms of increasing the separation resolution or for improvement of the detection. To increase the resolution, nanomaterials are employed as either surface modification of the capillary wall forming open tubular column or as additives to the separation electrolyte resulting in a pseudostationary phase. Moreover, nanomaterials have proven to be very beneficial for increasing also the sensitivity of detection employed in CE or even they enable the detection (e.g., fluorescent tags of nonfluorescent molecules).


Balloon Eustachian Tuboplasty in children.
Endonasal ballon dilatation of the Eustachian tube (BET) is a promising treatment for Eustachian tube dysfunction with encouraging results over the last years in adults. In addition, in children, single studies demonstrated promising results, but revealed the necessity for broader and additional studies. Our retrospective analysis presents outcomes with BET in children with chronic obstructive dysfunction of the Eustachian tube, showing resistance to the conventional therapy after adenotomy with paracentesis or grommets (ventilation tubes). The data of 52 children, having undergone BET from April 2011 to April 2016, were retrospectively evaluated. Most children in our study presented middle ear effusion (47%), adhesive (21%), chronic otitis media (13%), or recurrent acute otitis media (11%). In 24 (37%) children, we combined BET with a paracentesis, in 5 (8%) patients with a tympanoplasty type I and in 3 (5%) patients with a type III. All children were assessed using an audiogram, tympanometry, and tubomanometry (50 mbar) before and after BET. In addition, we evaluated the results of the Lübecker questionnaire, which we performed before and after BET. The childrens' ear-related and quality of life-related symptoms, such as pressure equalization, ear pressure, hearing loss, pain and limitation in daily life, and satisfaction pre- and postoperatively, were analyzed. In the majority of patients, we could see an improvement in the ear pressure, hearing loss, limitation in daily life, and satisfaction with recurrent inflammations, underlined by better outcomes in the tubomanometry and the tympanogram. BET in children is a safe, efficient, and promising method to treat chronic tube dysfunction, especially as a second line treatment, when adenotomy, paracentesis, or grommets failed before.


Comparison of ethanol production from corn cobs and switchgrass following a pyrolysis-based biorefinery approach.
One of the main obstacles in lignocellulosic ethanol production is the necessity of pretreatment and fractionation of the biomass feedstocks to produce sufficiently pure fermentable carbohydrates. In addition, the by-products (hemicellulose and lignin fraction) are of low value, when compared to dried distillers grains (DDG), the main by-product of corn ethanol. Fast pyrolysis is an alternative thermal conversion technology for processing biomass. It has recently been optimized to produce a stream rich in levoglucosan, a fermentable glucose precursor for biofuel production. Additional product streams might be of value to the petrochemical industry. However, biomass heterogeneity is known to impact the composition of pyrolytic product streams, as a complex mixture of aromatic compounds is recovered with the sugars, interfering with subsequent fermentation. The present study investigates the feasibility of fast pyrolysis to produce fermentable pyrolytic glucose from two abundant lignocellulosic biomass sources in Ontario, switchgrass (potential energy crop) and corn cobs (by-product of corn industry). Demineralization of biomass removes catalytic centers and increases the levoglucosan yield during pyrolysis. The ash content of biomass was significantly decreased by 82-90% in corn cobs when demineralized with acetic or nitric acid, respectively. In switchgrass, a reduction of only 50% for both acids could be achieved. Conversely, levoglucosan production increased 9- and 14-fold in corn cobs when rinsed with acetic and nitric acid, respectively, and increased 11-fold in switchgrass regardless of the acid used. After pyrolysis, different configurations for upgrading the pyrolytic sugars were assessed and the presence of potentially inhibitory compounds was approximated at each step as double integral of the UV spectrum signal of an HPLC assay. The results showed that water extraction followed by acid hydrolysis and solvent extraction was the best upgrading strategy. Ethanol yields achieved based on initial cellulose fraction were 27.8% in switchgrass and 27.0% in corn cobs. This study demonstrates that ethanol production from switchgrass and corn cobs is possible following a combined thermochemical and fermentative biorefinery approach, with ethanol yields comparable to results in conventional pretreatments and fermentation processes. The feedstock-independent fermentation ability can easily be assessed with a simple assay.


lncRNAs and MYC: An Intricate Relationship.
Long non-coding RNAs (lncRNAs) are emerging as important regulators of gene expression networks, acting either at the transcriptional level, by influencing histone modifications, or at the post-transcriptional level, by controlling mRNA stability and translation. Among the gene expression networks known to influence the process of oncogenic transformation, the one controlled by the proto-oncogene MYC is one of the most frequently deregulated in cancer. In B-cell lymphomas, the MYC gene is subject to chromosomal rearrangements that result in MYC overexpression. In many other cancers, the region surrounding MYC is subject to gene amplification. MYC expression is also controlled at the level of protein and mRNA stability. Neoplastic lesions affecting MYC expression are responsible for a drastic change in the number and the type of genes that are transcriptionally controlled by MYC, depending on differential promoter affinities. Transcriptome profiling of tumor samples has shown that several lncRNAs can be found differentially regulated by MYC in different cancer types and many of them can influence cancer cell viability and proliferation. At the same time, lncRNAs have been shown to be able to control the expression of MYC itself, both at transcriptional and post-transcriptional levels. Given that targeting the MYC-dependent transcriptional program has the potential to reach broad anticancer activity, molecular dissection of the complex regulatory mechanisms governing MYC expression will be crucial in the future for the identification of novel therapeutic strategies.


Contrast-enhanced ultrasound (CEUS) prediction of focal liver lesions in patients after liver transplantation in comparison to histopathology results.
To investigate the value of contrast-enhanced ultrasound (CEUS) in histologic prediction of focal liver lesions after liver transplantation. 10 focal liver lesions in 10 patients after liver transplantation were scanned using CEUS and the CEUS results were compared with the histopathological results. Among 10 focal liver lesions, 7 proofed to be histopathological benign and 3 lesions proofed to be histopathological malignant. All lesions (100%) were correctly report as benign or malignant in the report of the CEUS examination. CEUS can be helpful in the differentiation of benign and malignant focal liver lesions in patients after liver transplantation and can be used in clinical management of focal liver lesions.