Sequence capture-PCR improves detection of mycobacterial DNA in clinical specimens

The rapid identification of mycobacterial DNA in clinical samples by PCR can be useful in the diagnosis of tuberculous infections, but several large studies have found that the sensitivity of this approach is not better than that of culture. In order to improve the sensitivity of detection of mycobacterial DNA in clinical specimens from patients with paucibacillary forms of tuberculosis, we have developed a procedure permitting the specific capture of mycobacterial DNA in crude samples prior to amplification, thereby concentrating the target sequences and removing irrelevant DNA and other potential inhibitors of the amplification reaction (sequence capture-PCR). By using this approach to capture and amplify two different sequences specific for organisms of the Mycobacterium tuberculosis complex (IS6110 and the direct repeat region), it was possible to detect as little as one genome of mycobacterial DNA in samples containing up to 750 micrograms of total DNA, representing a 10- to 100-fold increase in sensitivity compared with that obtained by purifying total DNA prior to amplification. Detection of the IS6110 sequence in pleural fluid samples from patients with tuberculous pleurisy by sequence capture-PCR gave positive results in 13 of 17 cases, including 3 of 3 culture-positive samples and 10 of 14 culture-negative samples. In contrast, when total DNA was purified from these samples by adsorption to a silica matrix prior to amplification, only the three culture-positive samples were positive by PCR. The sensitivity of detection of the direct repeat sequence in these samples by sequence capture-PCR was similar to that of IS6110 and, in addition, permitted immediate typing of the strains from some patients. We conclude that sequence capture-PCR improves the sensitivity of detection of mycobacterial DNA in paucibacillary samples. This approach should be useful in detecting rare target sequences from organisms implicated in other pathologic processes.     

Nitric oxide reduction and carbon monoxide oxidation over carbon-supported copper-chromium catalysts

Carbon supported copper-chromium catalysts are shown to be very active for both the reduction of nitric oxide with carbon monoxide and the oxidation of carbon monoxide with oxygen. Mixed copper-chromium oxide active phases have good activity in the simultaneous removal of nitric oxide and carbon monoxide from exhaust gases. The influence of several catalyst variables has been investigated. The activity per volume of catalyst increases with increasing loading, while the intrinsic activity shows a maximum around C/M=100−50. An optimum catalyst for nitric oxide reduction and carbon monoxide oxidation has a copper/chromium ratio of 2/1. The apparent activation energy for the carbon monoxide oxidation over carbon supported copper-chromium catalysts is 77 kJ/mol, suggesting that the Cu---O bond rupture is the rate-limiting process. The reduction of nitric oxide takes place at higher temperatures. Since all catalysts have a low selectivity for molecular nitrogen formation at lower temperatures, the dissociation of nitric oxide is probably rate determining, resulting in a slightly reduced catalyst system. In an excess of carbon monoxide the reaction is first-order in nitric oxide and zero-order in carbon monoxide. Moisture inhibits the reaction by reversible competitive adsorption, whereas carbon dioxide does not. Oxygen completely inhibits the reduction of nitric oxide due to the more rapid reoxidation of the catalytic sites compared to nitric oxide. Therefore, the reduction of nitric oxide takes place only when all oxygen has been converted and, hence, is shifted to higher temperatures. As a possible consequence, the production of nitrous oxide is reduced. Nitric oxide and molecular oxygen react preferentially with carbon monoxide, so, in an excess of oxidizing component, gasification of the carbon support occurs at higher temperatures after carbon monoxide has been completely consumed.     

A novel cyclic process using CaSO<Subscript>4</Subscript>/CaS pellets for converting sulfur dioxide to elemental sulfur without generating secondary pollutants: Part I. Feasibility and kinetics of the reduction of sulfur dioxide with calcium-sulfide pellets

Certain new sulfide-smelting processes and coal-gasification processes generate high-strength sulfur dioxide streams, for which a new process for converting sulfur dioxide to elemental sulfur needs to be developed because no process exists that is generally and economically applicable to the treatment of such streams. A thermodynamic and experimental investigation to develop a new process for converting sulfur dioxide to elemental sulfur by a cyclic process involving calcium sulfide and calcium sulfate without generating secondary pollutants was carried out. In this process, the starting raw material, calcium sulfate, is reduced by a suitable reducing agent, such as hydrogen, to produce calcium sulfide, which is used to reduce sulfur dioxide to elemental sulfur vapor and calcium sulfate. The latter is, in turn, reduced to regenerate calcium sulfide. In this Part I, detailed experimental results are presented on the kinetics of the reaction between sulfur dioxide and calcium-sulfide pellets, which produces elemental sulfur and calcium sulfate. The experiments were carried out at temperatures between 1023 and 1088 K and sulfur-dioxide partial pressures between 9 and 60 kPa by the use of a thermogravimetric analysis (TGA) technique. The rate of this reaction was demonstrated by the conversion of 40 pct calcium-sulfide pellets obtained from the hydrogen reduction of fresh calcium sulfate in 10 minutes at 1073 K under a sulfur-dioxide partial pressure of 43 kPa. The reactivity decreased somewhat during the first three cycles but remained largely unchanged thereafter up to the tenth cycle. This characteristic of the pellets is important because the solids must be reusable for repeated cycles to avoid generating secondary pollutants. A pore-blocking model was found to fit the reaction rate. The reaction is first order with respect to sulfur-dioxide partial pressure and has an activation energy of 101 to 134 kJ/mol (24 to 32 kcal/mol) for calcium-sulfide pellets reacted and regenerated several different times. Sulfur dioxide-containing streams from certain sources, such as the regenerator off-gas from an integrated-gasification, combined-cycle, desulfurization unit and new sulfide-smelting plants, contain much higher partial pressures of SO₂. In these cases, the rate of the first reaction is expected to be proportionally higher than in the test conditions reported in this article. The reduction kinetics of calcium-sulfate pellets with hydrogen gas is reported in the accompanying Part II.     

Microbiological denitrification of red beet juice

The purpose of study was the determination of optimal conditions for red beet juice denitrification by Paracoccus denitrificans bacteria as well as the evaluation of the usefulness of microbiologically treated juice for the production of a natural colouring preparation for foods. Total reduction of nitrates was found during bacterial culturing at 25 and 30 °C, at an initial pH of 7.0-8.0 and juice osmotic pressure of 900-1100 mOsm/kg. Microbiological denitrification procedure affected colour, taste and aroma of the juice. However, the use of juice as food colouring and component was made feasible by acidification and partial evaporation of water together with volatile substances.     

Why is software late? An empirical study of reasons for delay insoftware development

A study of the reasons for delay in software development is described. The aim of the study was to gain an insight into the reasons for differences between plans and reality in development activities in order to be able to take actions for improvement. A classification was used to determine the reasons. 160 activities, comprising over 15000 hours of work, have been analyzed. The results and interpretations of the results are presented. Insight into the predominant reasons for delay enabled actions for improvements to be taken in the department concerned. Because the distribution of reasons for delay varied widely from one department to another, it is recommended that every department should gain an insight into its reasons for delay in order to be able to take adequate actions for improvement     

Efficient QoS support in a slotted multihop WDM metro ring

A novel distributed access protocol for a slotted wavelength-division-multiplexing (WDM) metro ring employing all-optical packet switching and supporting quality-of-service (QoS) classes is presented and analyzed. Since we assume that there are more nodes than available wavelengths in the network, we obtain a scalable multihop WDM ring as underlying network architecture. By dividing each channel into several time slots and further applying destination release and slot reuse, data packets can be efficiently transmitted and received in a statistically multiplexed manner. In our architecture, each node is equipped with one tunable transmitter and one fixed-tuned receiver. Furthermore, as we generally consider so-called a posteriori access strategies, different packet selection schemes are proposed and compared. An analytical model based on the semi-Markov process methodology is developed to quantify the performance of one of these schemes. As a key element of the protocol, an efficient QoS support access mechanism is proposed and its performance is evaluated. The new QoS control scheme adopts a frame-based slot reservation strategy including connection setup and termination, which only slightly increases the signaling and node processing overhead. Thus, an efficient hybrid protocol combining connectionless and connection-oriented packet transmissions is proposed     

Suspension force and mechanism of debris flows

The paper discusses the postulated suspension force, by which it is possible to explain some enigmatic phenomena observed in different types of mass movements. This force is generated in thick suspensions as a result of friction between sinking solid particles and static water. The suspension force is related to the seepage force but they have opposite directions. Both forces form flow networks. Having explained the suspension force, the paper describes how this influences the movement of material, particularly in debris flows. Enigmatic features and the mechanism of debris flows and lahars are explained by virtue of the suspension force. Electronic Publication