Methyltransferase Contingencies in the Pathway of Everninomicin D Antibiotics and Analogues

Everninomicins are orthoester oligosaccharide antibiotics with potent activity against multidrug-resistant bacterial pathogens. Everninomicins act by disrupting ribosomal assembly in a distinct region in comparison to clinically prescribed drugs. We employed microporous intergeneric conjugation with Escherichia coli to manipulate Micromonospora for targeted gene-replacement studies of multiple putative methyltransferases across the octasaccharide scaffold of everninomicin effecting the A₁, C, F, and H rings. Analyses of gene-replacement and genetic complementation mutants established the mutability of the everninomicin scaffold through the generation of 12 previously unreported analogues and, together with previous results, permitted assignment of the ten methyltransferases required for everninomicin biosynthesis. The in vitro activity of A₁- and H-ring-modifying methyltransferases demonstrated the ability to catalyze late-stage modification of the scaffold on an A₁-ring phenol and H-ring C-4’ hydroxy moiety. Together these results establish the potential of the everninomicin scaffold for modification through mutagenesis and in vitro modification of advanced biosynthetic intermediates.     

Bibliometric analysis of the evolution of biochar research trends and scientific production

Clean Technologies and Environmental Policy - Biochar has caught great attention over the last decade, yielding a large number of publications in a broad range of disciplines. This scientometric...     

Transforming the Danish Film Field Via ‘Professionalization’, Penetration and Integration

Using a qualitative case study of the transformations of the infrastructure of the Danish film field by the Danish Film Institute and its CEO, three basic questions about entrepreneurial agency are raised. The first deals with the relationship between the institutional environment and the personal backgrounds of entrepreneurial actors. The study finds that certain actors can be seen as possessing extreme or unique levels of legitimacy resulting in being rewarded with tremendous resources and opportunities for strategic action. The second question has to do with how, and what sort of role transformations can take place from a structurally central, but static position in a field. Here ‘intermediary entrepreneurialism’ was used to penetrate and integrate the field in such a way as to turn a structurally fixed position from a weak to a strong nexus. The third question deals with how transformations can be effected by an organization that straddles three prominent institutional fields and logics – the state, market and artistic expression. Here, the key role of the term ‘professionalization’ is emphasized, arguing that this term proved evocative and compelling in each institutional field. The case also adds to our understanding of institutional and transformation processes in creative industries by focusing on two less‐studied contexts – the role and strategies of centrally placed actors in initiating and leading transformational activities, and the role that entrepreneurial ‘bureaucrats’ can play in transforming creative industries.     

Biological evaluation of two anomeric glucose analogues iodinated in position 6

Two anomeric analogues of glucose labelled with 123 iodine in position 6, proposed as tracers of glucose transport in vivo, have been synthesized: alpha- and beta-methyl-6-deoxy-6-iodo-D-glucopyranoside (alpha MDIG and beta MDIG). The aim of this study was to determine whether these molecules interact with the glucose transporter and whether they could be used as tracers of glucose transport in vivo. The biodistribution of alpha MDIG and beta MDIG was studied in the mouse in vivo. To determine if these two anomers enter the cell via the glucose transporter, their uptake was measured in isolated perfused rat hearts, in human erythrocytes in suspension, and in cardiomyocytes of neonatal rat in culture. Both alpha MDIG and beta MDIG had similar repartitions in the mouse: myocardial uptake averaged 7% of the injected dose/g of organ at 2 min postinjection and alpha MDIG competed with D-glucose to enter the cells. Insulin produced a 123% increase of its uptake in isolated perfused rat hearts and a 100% increase in cardiomyocytes of neonatal rat in culture. alpha MDIG uptake was lowered in the presence of glucose transport inhibitors in each experimental model. An interaction between beta MDIG and glucose transporters was observed only in human erythrocytes in suspension. Only alpha MDIG interacts with the glucose transporter, and thus could be used to estimate glucose transport in vivo.     

Pulmonary function after one-lung ventilation in newborns: the basis for neonatal thoracoscopy

BACKGROUND: To maintain good exposure during major video-assisted thoracic surgery it is necessary to deflate completely the ipsilateral lung. However, little is known about the effects of one-lung ventilation (OLV) on pulmonary function in newborn patients. METHODS: Ten neonatal domestic pigs with a mean age of 6+/-0.6 days were intubated and ventilated in pressure-controlled mode (inspired oxygen fraction=1.0). One-lung ventilation was maintained for 120 minutes. Serial measurements of hemodynamics and gas exchange were done before, during, and until 90 minutes after OLV. Pulmonary function testing was performed before and after OLV for each lung separately. RESULTS: With the inspired oxygen fraction set at 1.0, arterial oxygen saturation remained stable at 100% during OLV. Venous admixture and alveolar-arterial oxygen tension gradient increased slightly from the baseline value of 2.6% +/-0.3% to 3.8%+/-0.3% during OLV (mean+/-standard error of the mean; p=0.02), and from 358+/-28 to 407+/-18 mm Hg (not significant), respectively. Both values returned to baseline during the subsequent ventilation of both lungs. Static compliance and resistance of the ventilated lung did not change. Compliance of the collapsed lung decreased after reexpansion from 0.42+/-0.07 to 0.29+/-0.06 mL x cm H2O(-1) x kg(-1), p=0.008). Resistance remained unchanged (0.22+/-0.02 versus 0.25+/-0.05 cm H2O x L(-1) x s(-1); not significant). CONCLUSIONS: There were only minor effects on pulmonary function during and after OLV in the neonatal piglet. Alterations in gas exchange during OLV were minimal. Prolonged collapse of the lung with subsequent reexpansion was associated with a slight decrease in compliance, indicating some mild lung injury.     

Phase Equilibria in the Na2O-Cs2O-SiO2 System

The Na₂O-Cs2O-SiO₂ system has been investigated by means of a new differential thermal analysis apparatus. Two compounds have been observed for the first time in the metasilicate and disilicate joins. The CsNaSi₂O₅ disilicate melts congruently at 1217 K and the peritectic fusion of the CsNaSiO₃ metasilicate occurs at 1120 K.     

Effects of magnetic resonance imaging on cardiac pacemakers and electrodes

In phantom studies we investigated the effects of magnetic resonance imaging (MRI) on pacemakers and electrodes. Twenty-five electrodes were exposed to MRI in a 1.5T scanner with continuous registration of the temperature at the electrode tip. Eleven pacemakers (five single chamber and six dual chamber) were exposed to MRI. Pacemaker output was monitored to detect malfunction in VOO/DOO and VVI/DDD modes. A temperature increase at the electrode tip of up to 63.1 degrees C was observed during 90 seconds of scanning. In seven electrodes the temperature increase exceeded 15 degrees C. Although no pacemaker malfunctions were observed in asynchronous pacing mode (VOO/DOO), inhibition and rapid pacing were observed during spin-echo imaging if the pacemakers were set to VVI or DDD mode. Pacemaker function was not impaired during scanning with gradient-echo sequences. Next to pacemaker dysfunction, electrode heating has to be considered a possible adverse effect when exposing patients with pacemakers to MRI.     

The occurrence of goitrogenic substances in milk. 1. Release of goitrin in the milk of cows fed on rapeseed extract cakes

A total of six cows, divided into 3 groups, were fed various amounts of rape cake containing 6 g of goitrin/kg over a period of 7 days. The cows were milked twice a day and the goitrin content of the heated milk samples were determined by a HPLC-method within 2 h. When rape cake was fed at 0.39, 1.9 and 3.9% resp. of the total feed this resulted in medium goitrin values of 37, 163 and 707 micrograms/l milk. These values correspond to a transfer of about 0.1% of the original progoitrin content in the feed. 12 h after the last rape feeding the amount of goitrin in the milk was below the detection limit of 7 ppb. The toxicological significance of these findings are dicussed.     

Aide in decision-making: contribution to uncertainties in three-dimensional measurement

The authorities of the standards organization International Organization of Standardization (ISO) advocate mastering any uncertainties in all parts of the industrialization process. In the three-dimensional (3D) measurement process, uncertainty is usually obtained at the end of a battery of tests. It is defined as a whole because it includes several types of errors, known systematic components, unknown systematic components and random components. Automated calculations of uncertainty can be made based on statistics. This method is based on statistical concepts, which are in accordance with “The Guide to the expression of the uncertainty in measurement” (GUM). It also enables us to generate uncertainties on the verification of ISO specifications (or specs in the ISO directives). In the course of this article, a usage will be presented that takes the knowledge of uncertainties into account: this usage will help the operator to take a decision on the conformance of a mechanical part in reference to its conformance to geometric tolerance.