Synthesis of Fluorine-Containing Analogues of Purine Deoxynucleosides: Optimization of Enzymatic Transglycosylation Conditions.

In this work, a comparative analysis of the conditions of transglycosylation reactions catalyzed by E. coli nucleoside phosphorylases was carried out, and the optimal conditions for the formation of various nucleosides were determined. Under the optimized conditions of transglycosylation reaction, fluorine-containing derivatives of N6-benzyl-2'-deoxyadenosine, potential inhibitors of replication of enteroviruses in a cell, were obtained starting from the corresponding ribonucleosides.

Are immunosuppressive conditions and preoperative corticosteroid treatment risk factors in inguinal hernia repair?

INTRODUCTION: Immunosuppressive conditions and/or preoperative corticosteroid treatment have a negative influence on wound healing and can, therefore, lead to higher rates of surgical site infections (SSIs) and seromas. For inguinal hernia, no such studies have been carried out to date. METHODS: In an analysis of data from the Herniamed Registry, 2312 of 142,488 (1.6%) patients with primary unilateral inguinal hernia repair had an anamnestic history of an immunosuppressive condition and/or preoperative corticosteroid treatment. Using propensity score matching, 2297 (99.4%) pairs with comparative patient characteristics were formed. These were then compared using the following primary outcome criteria: intra- and postoperative complications, complication-related reoperations, recurrence at one-year follow-up, pain on exertion, pain at rest, and chronic pain requiring treatment at one-year follow-up. Of the 2297 matched pairs with primary unilateral inguinal hernia repair, 82.76% were male patients. 1010 (44.0%) were operated in laparo-endoscopic techniques (TEP, TAPP), 1225 (53.3%) in open techniques (Bassini, Shouldice, Lichtenstein, Plug, TIP, Gilbert, Desarda), and 62 (2.7%) in other techniques. RESULTS: The matched pair analysis results did not identify any disadvantage in terms of the outcome criteria for patients with an anamnestic history of immunosuppressive condition and/or preoperative corticosteroid treatment (yes vs no). In particular, no disadvantage was noted in the rate of surgical site infections (0.65% vs 0.70%; ns) or seromas (1.22% vs 1.57%; ns). The overall rates of postoperative complications were 3.40% vs 4.31% (p = ns) (plus 0.22% concordant events in five matched pairs). CONCLUSION: In primary unilateral inguinal hernia surgery, an immunosuppressive condition and/or preoperative corticosteroid treatment does not appear to have a negative influence on wound complications.

Delayed Diagnoses of SGCE Myoclonus-Dystonia.

Background: Myoclonus-dystonia due to SGCE mutations (OMIM: 159900) most commonly presents during childhood with mainly upper body myoclonus, and mild dystonia affecting the neck and arms. Case reports: Herein, we report patients misdiagnosed during childhood with Tourette syndrome and dyskinetic cerebral palsy, and, during adulthood, found to harbor SGCE frameshift mutations. Discussion: Myoclonus-dystonia may be underdiagnosed due to phenotypic misclassification during childhood. SGCE mutations should be included in the differential diagnosis of childhood movement disorders that ostensibly manifest with tics, myoclonus, or abnormal posturing secondary to dystonia and/or spasticity. Highlights: Due to pleiotropy, variable penetrance, broad differential, and hereditary effects of imprinting, the diagnosis of a disorder of childhood onset, myoclonus-dystonia due to SGCE mutations, may be delayed until adulthood, often compromising appropriate clinical management and genetic counseling.

Comparison of efficacy of low-volume bowel cleansers prior to colonoscopy: a randomised, prospective, open-label trial.

INTRODUCTION: The aim of the study was to compare the efficacy and tolerability of polyethylene glycol/ascorbic acid (PEGA), sodium picosulfate/magnesium citrate (SPMC) and the oral sulfate formula (SIR) in a single- or split-dose regimen for bowel preparation prior to colonoscopy. METHODS: Randomised, multicentre, open-label study. The subjects received either PEGA, SPMC or SIR in the single- or split-dose regimen before the colonoscopy. Quality and tolerability of the preparation and complaints during preparation were recorded using a 5 point scale. RESULTS: 558 subject were analysed. Preparation quality was comparable in the single-dose regimen. The rate of satisfactory bowel cleansing (Aronchick score 1+2) was higher for split-dose SIR and PEGA compared to SPMC (95.6%, 86.2% vs. 72.5%, p.

Study of Ni-Catalyzed Graphitization Process of Diamond by in Situ X-ray Photoelectron Spectroscopy.

Graphene on diamond has been attracting considerable attention due to the unique and highly beneficial features of this heterostructure for a range of electronic applications. Here, ultrahigh-vacuum X-ray photoelectron spectroscopy is used for in situ analysis of the temperature dependence of the Ni-assisted thermally induced graphitization process of intrinsic nanocrystalline diamond thin films (65 nm thickness, 50-80 nm grain size) on silicon wafer substrates. Three major stages of diamond film transformation are determined from XPS during the thermal annealing in the temperature range from 300 °C to 800 °C. Heating from 300 °C causes removal of oxygen; formation of the disordered carbon phase is observed at 400 °C; the disordered carbon progressively transforms to graphitic phase whereas the diamond phase disappears from the surface from 500 °C. In the well-controllable temperature regime between 600 °C and 700 °C, the nanocrystalline diamond thin film is mainly preserved, while graphitic layers form on the surface as the predominant carbon phase. Moreover, the graphitization is facilitated by a disordered carbon interlayer that inherently forms between diamond and graphitic layers by Ni catalyst. Thus, the process results in formation of a multilayer heterostructure on silicon substrate.

Human cadavers preserved using Thiel's method for the teaching of fibreoptically-guided intubation of the trachea: a laboratory investigation.

We assessed the suitability of human cadavers preserved using Thiel's method for teaching flexible fibreoptic tracheal intubation. Thirty-one anaesthetists unacquainted with this technique received didactic teaching followed by handling of the fibrescope on the Oxford teaching box. They then carried out fibreoptic intubations in two cadavers to establish a baseline sample of their intubation skills. Thereafter, we randomly assigned the trainees to two groups to practice fibreoptic intubation either on two distinct cadavers or on two airway manikins. After 7 days we re-assessed procedural skills using the same cadavers as at baseline. Intubation time was the primary outcome and secondary outcomes included the incidence of failed intubations. We also evaluated trainee satisfaction. The mean (SD) intubation time decreased from a baseline value of 74 (20) s to 35 (6) s in the cadaver group and to 56 (16) s in the manikin group. The effect of 'time' was significant (p = 0.002), indicating that both methods of training led to improvements. The training effect of the cadaveric method was greater than with the manikin method (p = 0.0016). Thirty-four failed intubations occurred at baseline vs. eight at the end of study (RR 0.24, 95%CI 0.11-0.51, p = 0.0002, NNT 9.6); six in the cadaver group and two in the manikin group (p = 0.22). We conclude that human cadavers preserved using Thiel's method are potentially better for teaching flexible fibreoptic tracheal intubation compared with manikins.

Implementation of the European health interview survey (EHIS) into the German health update (GEDA).

BACKGROUND: This methodological paper describes the integration of the 'European Health Interview Survey wave 2' (EHIS 2) into the 'German Health Update' 2014/2015 (GEDA 2014/2015-EHIS). METHODS: GEDA 2014/2015-EHIS is a cross-sectional health survey. A two-stage stratified cluster sampling approach was used to recruit persons aged 15 years and older with permanent residence in Germany. Two different modes of data collection were used, self-administered web questionnaire and self-administered paper questionnaire. The survey instrument implemented the EHIS 2 modules on health status, health care use, health determinants and social background variables and additional national questions. Data processing was conducted according to the quality and validation rules specified by Eurostat. RESULTS: In total, 24,824 questionnaires were completed. The response rate was 27.6%. The two-stage cluster sample method seems to have been successful in achieving a sample with high representativeness. The final micro data file was inspected, approved and certified by Eurostat. Access to micro data of the EHIS 2 can be provided by Eurostat via research contract and to the GEDA 2014/2015-EHIS public use file by the Research Data Centre of the Robert Koch Institute. First EHIS 2 results are available at the Eurostat website. CONCLUSIONS: Integrating a multinational health survey into an existing national health monitoring system was a challenge in Germany. The national survey methodology for conducting the survey had to be further developed in order to meet the overarching goal of harmonizing the health information from national statistical offices and public health research institutes across the European Union. The harmonized EHIS 2 data source will profoundly impact international public health research in the near future. The next EHIS wave 3 will be conducted around 2019.

Silicon nanocrystal-based photonic crystal slabs with broadband and efficient directional light emission.

Light extraction from a thin planar layer can be increased by introducing a two-dimensional periodic pattern on its surface. This structure, the so-called photonic crystal (PhC) slab, then not only enhances the extraction efficiency of light but can direct the extracted emission into desired angles. Careful design of the structures is important in order to have a spectral overlap of the emission with extraction (leaky) modes. We show that by fabricating PhC slabs with optimized dimensions from silicon nanocrystals (SiNCs) active layers, the extraction efficiency of vertical light emission from SiNCs at a particular wavelength can be enhanced ∼ 11 times compared to that of uncorrugated SiNCs-rich layer. More importantly, increased light emission can be obtained in a broad spectral range and, simultaneously, the extracted light can stay confined within relatively narrow angle around the normal to the sample plane. We demonstrate experimentally and theoretically that the physical origin of the enhancement is such that light originating from SiNCs first couples to leaky modes of the PhCs and is then efficiently extracted into the surrounding.

Methods to Study Autophagy in Zebrafish.

Autophagy (cellular self-eating) is a highly regulated degradation process of the eukaryotic cell during which parts of the cytoplasm are delivered into, and broken down within, the lysosomal compartment. The process serves as a main route for the elimination of superfluous and damaged cellular constituents, thereby mediating macromolecular and organellar turnover. In addition to maintaining cellular homeostasis, autophagy is involved in various other cellular and developmental processes by degrading specific regulatory proteins, and contributing to the clearance of intracellular pathogens. The physiological roles and pathological involvement of autophagy can be effectively studied in divergent eukaryotic model systems ranging from yeast to mice. Such a tractable animal modelapplied only recently for autophagy researchis the zebrafish Danio rerio, which also facilitates the analysis of more specific biological processes such as tissue regeneration. In this chapter, we overview the main methods and tools that are used to monitor autophagic structures and to assay autophagic responses in this vertebrate organism. We place emphasis on genetic (functional) approaches applied for exploring novel cellular and developmental roles of the autophagic process.

Biosystem models, generated from a complex rule/reaction/influence network and from two functionality prototypes.

In this work we have further developed the Direct Computer Mapping (DCM) based modelling and simulation methodology. A unified, transition-based representation of complex rule, reaction and influence networks has been introduced and two prototypes (one general state- and another general transition-prototype) have been developed for the unified functional modelling of the state and transition nodes. Starting from the network and from the functional prototypes, an automatic generation method of the graphically editable and extensible GraphML description of biosystem models has been elaborated. The new developments have been implemented in the improved kernel of DCM models. The applied knowledge representation makes possible the unified generation and execution of the balance-based quantitative and influence- or rule-based qualitative, as well as optionally time-driven, multiscale biosystem models. Application of the developed methodology has been illustrated by the improved implementation of the formerly studied and upgraded example biosystem model for combining the detailed, quantitative p53/miR34a signalling system with the pathological model through an extended rule-based coupling model.

Measurement of capillary fragility: a useful tool to differentiate lipedema from obesity?

Lipedema is a disproportional obesity featuring spontaneous or light pressure-induce pain and frequent hematoma formation due to even minor traumatic injuries. It is generally distinguished from general obesity primarily based on clinical hallmarks; however, this becomes difficult when appearing in a concomitant form (combination of obesity and lipedema). Our study group has recently demonstrated that lipedema-associated bruising is correlated with increased capillary fragility (CF) and also that CF could be significantly improved by complex decongestive physiotherapy (CDP). In this study, we measured CF in female subjects with lipedema (15) or non-complicated obesity (15) who were body mass index (BMI) and waist-to-hip ratio (WHR) matched. CF was evaluated with the vacuum suction method (VSM) using Parrot's angiosterrometer in both groups. Application of VSM resulted in a significantly higher number of petechiae in subjects with lipedema. Capillary fragility measurement appears to be a useful differential diagnostic tool between lipedema and obesity under these trial parameters.

Cholecystectomy for Acute Cholecystitis After Renal Transplantation.

BACKGROUND: The aim of our study was to evaluate the rate of surgical complications, patient outcomes, and impact on graft function in renal transplant recipients in whom cholecystectomy for acute cholecystitis was performed. METHODS: We reviewed data on transplant patients from January 1, 2006, to December 31, 2013. The subgroup of patients who required subsequent cholecystectomy for acute cholecystitis was assessed, and their data were further analyzed. RESULTS: Thirty-one patients who underwent cholecystectomy for acute cholecystitis after renal transplantation were included in the study. Clinical signs such as pain in the right upper quadrant, temperature >38°C, and elevation in bilirubin levels occurred in 20 (64.5%), 8 (25.8%), and 3 (9.7%) patients, respectively. Ultrasound signs of acute cholecystitis were present in 27 patients (87.1%). In terms of laboratory values, white blood cell counts >10 × 10(9)/L occurred in 17 patients (54.8%), and C-reactive protein levels >40 mg/L were reported in 21 patients (67.7%). The conversion rate to open surgery was 32.3% (10 patients). In 13 cases, acalculous cholecystitis was present (41.9%). The average serum creatinine level 1 year after cholecystectomy had no statistically significant differences. One patient required temporary dialysis during the postoperative period (with subsequent graft recovery), and 1 graft was lost. CONCLUSIONS: Acute cholecystitis in kidney transplant recipients is a serious complication, with frequent difficulties related to evaluation and diagnosis. Because clinical signs could be very mild compared with severity of gallbladder affliction, there is little room if any for conservative treatment in these patients. We have not noticed adverse impact of acute cholecystitis on 1-year graft function.

Path Sampling Methods for Enzymatic Quantum Particle Transfer Reactions.

The mechanisms of enzymatic reactions are studied via a host of computational techniques. While previous methods have been used successfully, many fail to incorporate the full dynamical properties of enzymatic systems. This can lead to misleading results in cases where enzyme motion plays a significant role in the reaction coordinate, which is especially relevant in particle transfer reactions where nuclear tunneling may occur. In this chapter, we outline previous methods, as well as discuss newly developed dynamical methods to interrogate mechanisms of enzymatic particle transfer reactions. These new methods allow for the calculation of free energy barriers and kinetic isotope effects (KIEs) with the incorporation of quantum effects through centroid molecular dynamics (CMD) and the full complement of enzyme dynamics through transition path sampling (TPS). Recent work, summarized in this chapter, applied the method for calculation of free energy barriers to reaction in lactate dehydrogenase (LDH) and yeast alcohol dehydrogenase (YADH). We found that tunneling plays an insignificant role in YADH but plays a more significant role in LDH, though not dominant over classical transfer. Additionally, we summarize the application of a TPS algorithm for the calculation of reaction rates in tandem with CMD to calculate the primary H/D KIE of YADH from first principles. We found that the computationally obtained KIE is within the margin of error of experimentally determined KIEs and corresponds to the KIE of particle transfer in the enzyme. These methods provide new ways to investigate enzyme mechanism with the inclusion of protein and quantum dynamics.

Pathogenic Helicobacter pylori strains translocate DNA and activate TLR9 via the cancer-associated cag type IV secretion system.

Helicobacter pylori (H. pylori) is the strongest identified risk factor for gastric cancer, the third most common cause of cancer-related death worldwide. An H. pylori constituent that augments cancer risk is the strain-specific cag pathogenicity island, which encodes a type IV secretion system (T4SS) that translocates a pro-inflammatory and oncogenic protein, CagA, into epithelial cells. However, the majority of persons colonized with CagA+ H. pylori strains do not develop cancer, suggesting that other microbial effectors also have a role in carcinogenesis. Toll-like receptor 9 (TLR9) is an endosome bound, innate immune receptor that detects and responds to hypo-methylated CpG DNA motifs that are most commonly found in microbial genomes. High-expression tlr9 polymorphisms have been linked to the development of premalignant lesions in the stomach. We now demonstrate that levels of H. pylori-mediated TLR9 activation and expression are directly related to gastric cancer risk in human populations. Mechanistically, we show for the first time that the H. pylori cancer-associated cag T4SS is required for TLR9 activation and that H. pylori DNA is actively translocated by the cag T4SS to engage this host receptor. Activation of TLR9 occurs through a contact-dependent mechanism between pathogen and host, and involves transfer of microbial DNA that is both protected as well as exposed during transport. These results indicate that TLR9 activation via the cag island may modify the risk for malignancy within the context of H. pylori infection and provide an important framework for future studies investigating the microbial-epithelial interface in gastric carcinogenesis.

Suitability of a preserved human cadaver model for the simulation of facemask ventilation, direct laryngoscopy and tracheal intubation: a laboratory investigation.

BACKGROUND: Using fresh or formalin-embalmed cadavers has not been generally accepted for the purposes of teaching airway management. We investigated whether cadavers 'preserved according Thiel's embalming method' (PATEM) are suitable for the simulation of facemask ventilation and tracheal intubation by direct laryngoscopy. METHODS: This observational cluster sampling, controlled simulation study, included eight PATEM cadavers and eight manikins in two clusters. Twenty experienced anaesthetists were randomly assigned to execute 80 facemask ventilations and 80 tracheal intubations in both groups. The ease of facemask ventilation was the primary endpoint. The secondary endpoint was the composite outcomes of laryngoscopy and tracheal intubation. RESULTS: The success rate at the first attempt at mask ventilation was 74% (59/80 attempts) on cadavers and 41% (33/80 attempts) on manikins (P<0.0001). Twenty one subjects received an oral airway in both groups and succeeded in facemask ventilation 20 times on cadavers and four times on manikins (P=0.004). Two-handed technique mask ventilation was required 24 times on manikins and once on cadavers (P=0.0016). In one attempt on a manikin the mask ventilation was impossible. Poor laryngeal view (Cormack-Lehane grade 3) occurred 14 times among cadavers (17.5%) and once in manikins (1.25%) (P=0.007), whereas difficulties in tracheal intubation were encountered 16 times in cadavers (20%) vs 17 times in manikins (21.25%) (P=0.84). In a subjective evaluation the participants preferred the cadaver model over the manikins (P<0.0001). CONCLUSIONS: PATEM cadavers were better suited for facemask ventilation and provided a more realistic environment for laryngoscopy and tracheal intubation than the studied manikins.

A New Surgical Model of Skeletal Muscle Injuries in Rats Reproduces Human Sports Lesions.

Skeletal muscle injuries are the most common sports-related injuries in sports medicine. In this work, we have generated a new surgically-induced skeletal muscle injury in rats, by using a biopsy needle, which could be easily reproduced and highly mimics skeletal muscle lesions detected in human athletes. By means of histology, immunofluorescence and MRI imaging, we corroborated that our model reproduced the necrosis, inflammation and regeneration processes observed in dystrophic mdx-mice, a model of spontaneous muscle injury, and realistically mimicked the muscle lesions observed in professional athletes. Surgically-injured rat skeletal muscles demonstrated the longitudinal process of muscle regeneration and fibrogenesis as stated by Myosin Heavy Chain developmental (MHCd) and collagen-I protein expression. MRI imaging analysis demonstrated that our muscle injury model reproduces the grade I-II type lesions detected in professional soccer players, including edema around the central tendon and the typically high signal feather shape along muscle fibers. A significant reduction of 30% in maximum tetanus force was also registered after 2 weeks of muscle injury. This new model represents an excellent approach to the study of the mechanisms of muscle injury and repair, and could open new avenues for developing innovative therapeutic approaches to skeletal muscle regeneration in sports medicine.

Is ionized oxygen negatively or positively charged more effective for carboxyhemoglobin reduction compare to medical oxygen at atmospheric pressure?

Carbon monoxide (CO) reversibly binds to hemoglobin forming carboxyhemoglobin (COHb). CO competes with O(2) for binding place in hemoglobin leading to tissue hypoxia. Already 30 % saturation of COHb can be deadly. Medical oxygen at atmospheric pressure as a therapy is not enough effective. Therefore hyperbaric oxygen O(2) inhalation is recommended. There was a question if partially ionized oxygen can be a better treatment at atmospheric pressure. In present study we evaluated effect of partially ionized oxygen produced by device Oxygen Ion 3000 by Dr. Engler in elimination of COHb in vitro experiments and in smokers. Diluted blood with different content of CO was purged with 5 l/min of either medicinal oxygen O(2), negatively ionized O(2) or positively ionized O(2) for 15 min, then the COHb content was checked. In vivo study, 15 smokers inhaled of either medicinal oxygen O(2) or negatively ionized O(2), than we compared CO levels in expired air before and after inhalation. In both studies we found the highest elimination of CO when we used negatively ionized O(2). These results confirmed the benefit of short inhalation of negatively ionized O(2), in frame of Ionized Oxygen Therapy (I O(2)Th/Engler) which could be used in smokers for decreasing of COHb in blood.