An efficient method for the synthesis of peptide aldehyde libraries employed in the discovery of reversible SARS coronavirus main protease (SARS-CoV Mpro) inhibitors

A method for the parallel solid-phase synthesis of peptide aldehydes has been developed. Protected amino acid aldehydes obtained by the racemization-free oxidation of amino alcohols with Dess-Martin periodinane were immobilized on threonyl resins as oxazolidines. Following Boc protection of the ring nitrogen to yield the N-protected oxazolidine linker, peptide synthesis was performed efficiently on this resin. A peptide aldehyde library was designed for targeting the SARS coronavirus main protease, SARS-CoV M(pro)(also known as 3CL(pro)), on the basis of three different reported binding modes and supported by virtual screening. A set of 25 peptide aldehydes was prepared by this method and investigated in inhibition assays against SARS-CoV M(pro). Several potent inhibitors were found with IC(50) values in the low micromolar range. An IC(50) of 7.5 muM was found for AcNSTSQ-H and AcESTLQ-H. Interestingly, the most potent inhibitors seem to bind to SARS-CoV M(pro) in a noncanonical binding mode.     

Experimental validation of a geographical information systems-based procedure for predicting pesticide exposure in surface water

A GIS-based procedure for predicting pesticide exposure in surface waters has been applied on a pilot river basin characterized by intensive agricultural activity. The predictive approach has been validated through experimental monitoring, performed by collecting manual and automatic water samples during the productive season. Five active ingredients (terbuthylazine, metolachlor, alachlor, linuron, fenitrothion) were selected for analysis to validate the predictive approach. Comparison between predicted and experimental values showed good agreement for terbuthylazine and metolachlor (used in large volumes within the basin), demonstrating the reliability of the approach. However, some anomalous results were obtained for some of the other chemicals, which serve to highlight the difficulties in getting reliable input data, in particular on application patterns (rate and time). Furthermore, the value of mapping pesticide exposure on the medium-large scale is described, and the limitations of the reported predictive approach are discussed.     

FEMMINA real-time, radio-frequency echo-signal equipment for testing novel investigation methods

Fast echographic multiparameter multi-image novel apparatus (FEMMINA), is a hardware and software platform dedicated to ultrasonic signal and image processing. FEMMINA is able to operate with sequences of radiofrequency (RF) frames. Its architecture is designed to be modular, expandable, and aimed at implementing different ultrasonic investigation techniques. The first experimental characteristic of this system is in its capability to operate in real time with ultrasonic RF signals, starting from acquisition up to processing, storage, and visualization. The second characteristic is the user-system interactivity that allows one to modify the operation appropriately while observing results. Currently, FEMMINA works in both typical experimental situations to study novel investigation techniques and clinical field to validate the proposed methods in different human districts.     

Power efficiency evaluation in Dickson and voltage doubler charge pump topologies

This paper presents a theoretical and experimental comparison between two charge pump architectures commonly used in CMOS integrated circuits, namely the Dickson scheme and the cascade of voltage doublers. The comparison is carried out considering power efficiency as the main feature of interest. To compare the two topologies, two charge pumps were integrated in 0.18-μm triple-well CMOS technology. The two charge pumps were designed with the same operating clock frequency, the same storage capacitance per stage, and the same number of stages (and, thus, approximately the same area). The theoretical and the experimental comparison showed that the power efficiency of the voltage doubler scheme is higher (by about 13% at ), mainly thanks to the lower parasitic capacitance associated to the boosted nodes.     

Sample treatment considerations in the analysis of organic colorants by surface-enhanced Raman scattering

The introduction of surface-enhanced Raman spectroscopy (SERS) in the field of cultural heritage has significantly improved the analysis of the organic dyes and their complexes that have been used as textile dyes and pigments in paintings and other polychrome works of art since antiquity. Over the last five years, a number of different procedures have been developed by various research groups. In this Article, we evaluate the effect of pretreating samples by exposing them to hydrofluoric acid (HF) vapor prior to SERS analysis, a step designed to hydrolyze the dye-metal complexes and increase analyte adsorption on the nanosized metallic support, thus enhancing the SERS signal. Materials studied include pure colorants, commercial lake pigments, and fibers from dyed textiles, as well as actual aged samples, such as microscopic fragments of lakes on paper and ancient pigments and glazes from several works of art, covering a wide range of time, from the second century B.C. to the early 20th century. In each case, SERS spectra obtained with or without HF hydrolysis were critically evaluated. The pretreatment with HF vapor resulted in faster analysis and increased sensitivity in most cases, with the exception of dyed silk fibers, where silk protein hydrolyzates were found to interfere with SERS analysis. As a final point, a two-step procedure including SERS on untreated and treated samples is proposed as a standard approach: by analyzing a sample first without hydrolysis, and then, following removal of the colloid, upon HF treatment, the best and most reliable results for a great number of dyes and substrates are assured.     

Construction of xylose dehydrogenase displayed on the surface of bacteria using ice nucleation protein for sensitive D-xylose detection

A novel method was developed to detect d-xylose (INS 967) sensitively and selectively, which is based on a xylose dehydrogenase (XDH) cell-surface displaying system using a newly identified ice nucleation protein from Pseudomonas borealis DL7 as an anchoring motif. With coenzyme NAD(+), the XDH-displayed bacteria facilitates the catalysis of the oxidization of xylose and the resultant NADH can be detected spectrometrically at 340 nm. The fusion protein was characterized by proteinase accessibility, Western blot, and enzyme activity assays. The established XDH surface displaying system did not inhibit the growth of the recombinant Escherichia coli strain. The XDH was mainly displayed on the surface of host cells, which is of high XDH activity and high D-xylose specificity. The optimal temperature and pH of cell displayed XDH were found at 30 °C and pH 8.0, respectively. The XDH-displayed bacteria can be used directly without further enzyme extraction and purification, and it improved the stability of the enzyme. Moreover, the cell-surface-displayed-protein-based approach showed a wide linear range (5-900 μM) and a low detection limit of 2 μM of d-xylose. More importantly, the recombinant cells could be used for precise detection of D-xylose from the real samples such as foods and degradation products of lignocellulose. The method shown here provides a simple, rapid, and low-cost strategy for the sensitive and selective measurement of D-xylose. In addition, the XDH-displayed bacteria showed an interesting response in developing electrochemical biosensors. Thus, the genetically engineered cells may find broad application in such biosensors and biocatalysts. Similarly, this type of genetic approach may be used for the expression of other intracellular enzymes of interest for certain purposes.     

How To Find a Needle (or Anything Else) in a Haystack: Two-Dimensional Correlation Spectroscopy-Filtering with Iterative Random Sampling Applied to Pharmaceutical Heparin

Risks of contamination of the major clinical anticoagulant heparin can arise from deliberate adulteration with unnatural or natural polysaccharides, including heparin from other animal sources, other natural products, or artifacts of manufacture, and these can escape detection by conventional means. Currently, there is no generally applicable, objective test recommended by regulators that can detect these in pharmaceutical heparin, and this continues to leave heparin exposed to contamination risks. Two-dimensional correlation spectroscopic-filtering with iterative random sampling (2D-COS-firs) is reported. It employs a difference covariance matrix with iterative random sampling, and is capable of revealing contamination in pharmaceutical heparin to a high level of sensitivity irrespective of the nature of those features. The technique is suitable to any situation in which a comparison of a single entity to a family of heterogeneous entities, particularly natural products and biosimilars, needs to be made, and will find application in pharmaceutical monitoring, manufacturing quality control, materials science, biotechnology, and metabolomic investigations.     

Formulating and optimizing a combustion pathways for oil shale and its semi-coke

Enhanced technologies from oil recovery to unconventional fuels - oil shale, oil sands and extra-heavy oil – have in common complex chemical reactions processes. This paper is about the formulation and optimization of the chemical mechanism especially in oil shale and semi-coke combustion. The Levenberg–Marquardt algorithm was used to minimize the error between estimated values and the thermogravimetric data for combustion mechanisms of 4-steps and 3-steps proposed for the oil shale and its semi-coke respectively. The kinetic parameters such as reaction order, pre-exponential factor, activation energy and stoichiometric coefficients that affect drying, pyrolysis, oxidation and decarbonation reactions were estimated with success. The values of activation energies were 54–67 kJ mol^?1 for oil shale drying, 62–65 kJ mol^?1 for pyrolysis reaction, up to 100 kJ mol^?1 for Fixed Carbon (FC) oxidation reaction, and 162–418 kJ mol^?1 for decarbonation reaction. Regarding to the semi-coke combustion, the activation energies were 33 kJ mol^?1 for drying reaction, 211 kJ mol^?1 for oxidation reaction and 291 kJ mol^?1 for decarbonation reaction. The chemical reactions suggest reaction order superior to one, except to the decarbonation reaction at 3 K min^?1. Considering the estimated parameters, as well as a heating rate at 3 K min^?1, an oil shale containing about 20 wt.% of organic matter and 34.6 wt.% of CaCO₃, the species mass fractions formed during combustion process were 3.4 wt.% of FC, 10.6 wt.% of Oil, 3.3 wt.% of HC and 1.8 wt.% of CO. The fraction of CO₂ formed accounts a total of 21.6 wt.%. For a semi-coke containing 3.4 wt.% of FC and 40.6 wt.% of CaCO₃, its combustion formed 2.1 wt.% of CO. The CO₂ fraction from oxidation and decarbonation reactions accounts 10.2 wt.%, considering that the stoichiometric mass coefficient γ = 0.75 in decarbonation reaction.     

Combination of electrophoretic deposition and microwave-ignited combustion synthesis for the preparation of ceramic coated intermetallic-based materials

Electrophoretic deposition (EPD) was used to deposit sub-micrometric ZrO₂ particles on metallic powder compacts belonging to the systems Ni + Al and Ti + Al, which were used as deposition electrodes in the EPD cell. After EPD, combustion synthesis (CS) of such reactive electrodes was ignited in a microwave single-mode applicator, operating at a frequency of 2.45 GHz, in order to obtain in a single step the synthesis of the desired intermetallic phase (substrate) and the sintering of the previously deposited ceramic particles (or coating). Experimental results demonstrate that the excess heat released during the formation of nickel and titanium aluminides by CS can be exploited not only to self-sustain and self-propagate the reaction front along the substrate, but also to rapidly sinter the coating obtained by EPD. The innovative procedure here proposed is a promising strategy in order to obtain, in a single step, high temperature intermetallic-based materials, protected by well adhered ceramic coatings.     

Survey of the interventional cardiology procedures in Italy

Interventional cardiology procedures are increasing because they offer many advantages to patients compared with other techniques: therefore the Italian National Institution for Insurance against Accidents at Work decided to start a survey for monitoring the state-of-the-art regarding the professionals involved in those procedures. The survey covered six cardiology and medical physics Italian departments. Each centre was asked to record 10 examinations for five types of procedures: coronary angiography (CA), electrophysiology studies (ES), pacemaker implantation (PI), percutaneous transluminal coronary angioplasty (PTCA) and radiofrequency catheter ablation (RA). For each examination all the centres were requested to fill in a questionnaire containing information regarding the operator performing the examination, the patient and the procedure. A total of 290 examinations were recorded: 103 CA, 14 ES, 68 PI, 79 PTCA and 26 RA. As occupational doses are strongly related to patient doses, both patients and operators radiation dose data are reported. Ratios of maximum to minimum mean patient doses across the hospitals surveyed were 2.0, 3.9, 7.0, 1.8 and 1.4 for CA, ES, PI, PTCA and RA, respectively. The calculated rounded mean dose-area product values across all participating hospitals were comparable with other values reported in the literature. In general, specific radiation protection tools were used by all operators performing different procedures in all hospitals. A major issue in this survey was the absence of information about correlation between staff and patient doses in a single procedure: future studies could be more aimed to prospective goals where occupational exposures per procedure are monitored specifically.