Looking at the future of manufacturing metrology: roadmap document of the German VDI/VDE Society for Measurement and Automatic Control

"Faster, safer, more accurately and more flexibly'' is the title of the "manufacturing metrology roadmap'' issued by the VDI/VDE Society for Measurement and Automatic Control (<a href="http://www.vdi.de/gma"target="_blank">http://www.vdi.de/gma</a>). The document presents a view of the development of metrology for industrial production over the next ten years and was drawn up by a German group of experts from research and industry. The following paper summarizes the content of the roadmap and explains the individual concepts of "Faster, safer, more accurately and more flexibly'' with the aid of examples

Detection of respiratory bacterial pathogens causing atypical pneumonia by multiplex Lightmix^® RT-PCR.

Pneumonia is a severe infectious disease. In addition to common viruses and bacterial pathogens (e.g. Streptococcus pneumoniae), fastidious respiratory pathogens like Chlamydia pneumoniae, Mycoplasma pneumoniae and Legionella spp. can cause severe atypical pneumonia. They do not respond to penicillin derivatives, which may cause failure of antibiotic empirical therapy. The same applies for infections with B. pertussis and B. parapertussis, the cause of pertussis disease, that may present atypically and need to be treated with macrolides. Moreover, these fastidious bacteria are difficult to identify by culture or serology, and therefore often remain undetected. Thus, rapid and accurate identification of bacterial pathogens causing atypical pneumonia is crucial. We performed a retrospective method evaluation study to evaluate the diagnostic performance of the new, commercially available Lightmix &lt;sup&gt;®&lt;/sup&gt; multiplex RT-PCR assay that detects these fastidious bacterial pathogens causing atypical pneumonia. In this retrospective study, 368 clinical respiratory specimens, obtained from patients suffering from atypical pneumonia that have been tested negative for the presence of common agents of pneumonia by culture and viral PCR, were investigated. These clinical specimens have been previously characterized by singleplex RT-PCR assays in our diagnostic laboratory and were used to evaluate the diagnostic performance of the respiratory multiplex Lightmix &lt;sup&gt;®&lt;/sup&gt; RT-PCR. The multiplex RT-PCR displayed a limit of detection between 5 and 10 DNA copies for different in-panel organisms and showed identical performance characteristics with respect to specificity and sensitivity as in-house singleplex RT-PCRs for pathogen detection. The Lightmix &lt;sup&gt;®&lt;/sup&gt; multiplex RT-PCR assay represents a low-cost, time-saving and accurate diagnostic tool with high throughput potential. The time-to-result using an automated DNA extraction device for respiratory specimens followed by multiplex RT-PCR detection was below 4 h, which is expected to significantly improve diagnostics for atypical pneumonia-associated bacterial pathogens

QUIRMIA-A Phenotype-Based Algorithm for the Inference of Quinolone Resistance Mechanisms in Escherichia coli

OBJECTIVES: Quinolone resistance in Escherichia coli occurs mainly as a result of mutations in the quinolone-resistance-determining regions of gyrA and parC, which encode the drugs' primary targets. Mutational alterations affecting drug permeability or efflux as well as plasmid-based resistance mechanisms can also contribute to resistance, albeit to a lesser extent. Simplifying and generalizing complex evolutionary trajectories, low-level resistance towards fluoroquinolones arises from a single mutation in gyrA, while clinical high-level resistance is associated with two mutations in gyrA plus one mutation in parC. Both low- and high-level resistance can be detected phenotypically using nalidixic acid and fluoroquinolones such as ciprofloxacin, respectively. The aim of this study was to develop a decision tree based on disc diffusion data and to define epidemiological cut-offs to infer resistance mechanisms and to predict clinical resistance in E. coli. This diagnostic algorithm should provide a coherent genotype/phenotype classification, which separates the wildtype from any non-wildtype and further differentiates within the non-wildtype. METHODS: Phenotypic susceptibility of 553 clinical E. coli isolates towards nalidixic acid, ciprofloxacin, norfloxacin and levofloxacin was determined by disc diffusion, and the genomes were sequenced. Based on epidemiological cut-offs, we developed a QUInolone Resistance Mechanisms Inference Algorithm (QUIRMIA) to infer the underlying resistance mechanisms responsible for the corresponding phenotypes, resulting in the categorization as "susceptible" (wildtype), "low-level resistance" (non-wildtype) and "high-level resistance" (non-wildtype). The congruence of phenotypes and whole genome sequencing (WGS)-derived genotypes was then assigned using QUIRMIA- and EUCAST-based AST interpretation. RESULTS: QUIRMIA-based inference of resistance mechanisms and sequencing data were highly congruent (542/553, 98%). In contrast, EUCAST-based classification with its binary classification into "susceptible" and "resistant" isolates failed to recognize and properly categorize low-level resistant isolates. CONCLUSIONS: QUIRMIA provides a coherent genotype/phenotype categorization and may be integrated in the EUCAST expert rule set, thereby enabling reliable detection of low-level resistant isolates, which may help to better predict outcome and to prevent the emergence of clinical resistance

Interventions for erythema nodosum leprosum

Background Erythema nodosum leprosum (ENL) is a serious immunological complication of leprosy, causing inflammation of skin, nerves, other organs, and general malaise. Many different therapies exist for ENL, but it is unclear if they work or which therapy is optimal. Objectives To assess the effects of interventions for erythema nodosum leprosum. Search strategy We searched the Cochrane Skin Group Specialised Register, the Cochrane Central Register of Controlled Trials (CENTRAL) in The Cochrane Library (Issue 1, 2009), MEDLINE (from 2003), EMBASE (from 2005), LILACS and AMED (from inception), CINAHL (from 1981), and databases of ongoing trials, all in March 2009. We checked reference lists of articles and contacted the American Leprosy Missions in Brazil to locate studies. Selection criteria Randomised controlled trials (RCTs) of interventions for ENL in people with leprosy. Data collection and analysis Two authors performed study selection, assessed trial quality, and extracted data. Main results We included 13 studies with a total of 445 participants. The quality of the trials was generally poor and no results could be pooled due to the treatments being so heterogeneous. Treatment with thalidomide showed a significant remission of skin lesions compared to acetylsalicylic acid (aspirin) (RR 2.43; 95% CI 1.28 to 4.59) (1 trial, 92 participants). Clofazimine treatment was superior to prednisolone (more treatment successes; RR 3.67; 95% CI 1.36 to 9.91) (1 trial, 24 participants), and thalidomide (fewer recurrences; RR0.08; 95% CI 0.01 to 0.56) (1 trial, 72 participants). We did not find any significant benefit for intravenous betamethasone compared to dextrose (1 trial, 10 participants), pentoxifylline compared to thalidomide (1 trial, 44 participants), indomethacin compared to prednisolone, aspirin or chloroquine treatments (2 trials, 80 participants), or levamisole compared to placebo (1 trial, 12 participants). Mild to moderate adverse events were significantly lower in participants taking 100 mg thalidomide compared to 300 mg thalidomide daily (RR 0.46; 95% CI 0.23 to 0.93). Significantly more minor adverse events were reported in participants taking clofazimine compared with prednisolone (RR 1.92; 95% CI 1.10 to 3.35). None of the studies assessed quality of life or economic outcomes. Authors' conclusions There is some evidence of benefit for thalidomide and clofazimine, but generally we did not find clear evidence of benefit for interventions in the management of ENL. However, this does not mean they do not work, because the studies were small and poorly reported. Larger studies using clearly defined participants, outcome measures, and internationally recognised scales are urgently required

The diagnosis of leprosy - common errors

Diagnosis of leprosyWould the correct diagnosis of leprosy have been easier if this disease had been described in textbooks on Neurology instead of Dermatology? The emphasis on the changes in the skin of the patient with leprosy may well be the cause of misdiagnosis in many cases. Leprosy is a chronic infective disease and it is generally accepted to be caused by Mycobacterium Leprae discovered by Dr. A. Hansen in 1873 and published by him in 1874. M. Leprae is an acid-alcohol fast bacillus. So far no artificial medium has been found in which to culture the bacillus, but it can be kept alive and will multiply in the earholes and foot pads of the mouse and hamster. The foot pads are used solely as a culture medium enabling the testing of the effect of drugs on the bacillus. It is important to know that M. Leprae has a special affinity for the Schwann cells of the sensory nerves in which they lie, protected by the basement membrane, (only seen by electron microscopy) and if conditions are suitable multiply. Only nerves of the Peripheral Nervous system are affected in leprosy. Therefore without symptoms showing nerve involvement the diagnosis of leprosy should not be made in the absence of positive skin smears. The great auricular nerve-the ulner and median and peroneal nerves are easy to palpate and possess predominantly cutaneous sensory fibres. In the early stages of the disease the bacillary invasion is directed to the sensory fibres while later on all types of fibres are affected due to granulomatous infiltration, scarring, ischaemic damage or even possibly tramsneuronal spread. Autonomic nerve fibres are commonly involved early in the disease, shown by the characteristic dryness and roughness of the skin and anhydrosis. This article is mainly written for doctors who have never worked in countries where leprosy is prevalent and therefore have some difficulty in recognising the disease. To be Leprosy-conscious is as important and essential as to be Malaria-conscious and this applies to patients of all races. Leprosy can occur in ALL races and at any age. It should bc born in mind that Europeans are not exempt from leprosy and unfortunately it has happened that Europeans have been treated for many years for an unidentified skin disease which later proved to be leprosy.Office of Global AIDS/US Department of State

Energy conservation in acetogenic bacteria : caffeate reduction in Acetobacterium woodii

Aus Fruktose-gezogenen Zellen von A. woodii, die in Gegenwart von Caffeat als Elektronenakzeptor gewachsen waren, wurde durch Behandlung mit Lysozym und anschliessendem French-Press-Aufschluß bei niedrigem Druck ein zellfreier Rohextrakt unter strikt anaeroben Bedingungen hergestellt. Dieser katalysierte eine H2-abhängige Caffeatreduktion mit Raten von 8,7 – 18,7 nmol/min x mg Protein. Die Aktivität war strikt ATP-abhängig. Nach einer Auftrennung des Rohextraktes in Cytoplasma- und Membranfraktion war die H2-abhängige Caffeatreduktion ausschliesslich in der cytoplasmatischen Fraktion lokalisiert. Die Zugabe von Membranen führte zu keiner Stimulierung der Aktivität. Die Membranfraktion selbst wies keine Caffeatreduktionsaktivität auf. Verschiedene Verbindungen wurden auf ihre Fähigkeit getestet, als artifizielle Elektronendonatoren für die Caffeatreduktion zu fungieren. TMPD, 1,5-Diphenylcarbazid und reduziertes Methylviologen konnten Caffeat nicht reduzieren. In Gegenwart von Phenylendiamin wurde in zellfreiem Rohextrakt und in der Cytoplasmafraktion Caffeatreduktionsaktivität beobachtet. In der Membranfraktion wurde dagegen keine Reduktion von Caffeat mit Phenylendiamin als Elektronendonor nachgewiesen. NADH + H+ konnte als physiologischer Elektronendonor für die Reduktion von Caffeat fungieren. Die NADH-abhängige Caffeatreduktion war ausschliesslich in der cytoplasmatischen Fraktion lokalisiert und strikt abhängig von ATP. Die Beobachtung, dass NADH + H+ als Elektronendonor für die Caffeareduktion fungieren kann, führte zu der Frage, wie im Zuge H2-abhängiger Caffeatreduktion NADH + H+ gebildet wird und welche Enzyme daran beteiligt sein könnten. NAD+-abhängige Hydrogenaseaktivität wurde an Membranen und im Cytoplasma nachgewiesen. Rund 70% der Aktivität waren in der löslichen Fraktion lokalisiert. Die Gegenwart einer Elektronendonor:NAD+-Oxidoreduktase in A. woodii wurde untersucht. Gewaschene Membranen vermittelten die Oxidation von NADH + H+ mit Kaliumhexacyanoferrat oder Benzylviologen als Elektronenakzeptor. Diese Beobachtung wurde als Hinweis auf eine NAD+-Reduktase gewertet, da solche Enzyme in der Regel reversibel sind. Eine Hydrogenase konnte hierfür ausgeschlossen werden, da die NADH-oxidierende Aktivität Sauerstoff-unempfindlich war. Die Aktivität des NADH-oxidierenden Enzyms konnte durch zugesetztes Na+ nicht stimuliert werden. In Analogie zu Na+-translozierenden NADH:Chinon-Reduktasen wurde die NADH-oxidierende Aktivität an gewaschenen Membranen aber durch Ag+ oder Cu2+ in mikromolaren Konzentrationen vollständig inhibiert. Membranen von A. woodii vermittelten die Reduktion von NAD+ mit reduzierten Ferredoxin als Elektronendonor. Ob diese Aktivität durch das gleiche membranständige Enzyme katalysiert wurde, das auch die NADH-Oxidation bewerkstelligte, konnte nicht geklärt werden. Die Ferredoxin-abhängige NAD+-Reduktion wurde durch micromolare Konzentrationen Ag+ vollständig inhibiert. Die Inihibition war aber wahrscheinlich unspezifischer Natur. Mittels vergleichender 2D-Gelelektrophorese wurden zwei Caffeat-induzierte Proteine identifiziert. Ein Vergleich der Peptidsequenzen, die durch ESI-MS/MS-Analyse der Proteine erhalten wurden, mit in Datenbanken hinterlegten Proteinsequenzen, ergab eine große Übereinstimmung zu der großen alpha- bzw. kleinen beta-Untereinheit von heterodimeren Elektronentransfer-Flavoproteinen (ETFP). Die Proteine wurden mit EtfA und EtfB bezeichnet. Anhand der Peptidsequenzen von EtfA und EtfB wurden degenerierte Oligonukleotide abgeleitet, die zur Amplifikation von Fragmenten der kodierenden Gene herangezogen wurden. Die Analyse der abgeleiteten Aminosäuresequenzen der erhaltenen PCR-Produkte untermauerte die Zuordnung von EtfA und EtfB als Untereinheiten eines ETFP. Die Fähigkeit zur Caffeatreduktion ist in A. woodii nicht konstitutiv vorhanden, sondern wird in erst durch Gegenwart des Phenylacrylats induziert. Die Spezifität dieser Induktion wurde untersucht. Suspensionen ruhender Zellen, die aus Caffeat-induzierten Zellen hergestellt worden waren, reduzierten neben Caffeat auch die Phenylacrylate Ferulat oder p-Cumarsäure mit H2 als Elektronendonor. Analoge Beoabachtungen wurden mit Ferulat-induzierten und p-Cumarsäure-induzierten Zellsuspensionen gemacht. Die Ergebnisse legen den Schluß nahe, dass in A. woodii die Reduktion von Phenylacrylaten durch ein induzierbares enzymatisches System bewerkstelligt wird. EtfA und EtfB wurden als MalE-Fusionsproteine dargestellt und gereinigt. Dagegen wurden Antiseren hergestellt. Immunologische Untersuchungen zeigten, dass die Produktion von EtfA und EtfB durch Gegenwart verschiedener Phenylacrylate induziert wird. Die Induktion war unabhängig vom Wachstumssubstrat. In Gegenwart von zu Phenylacrylaten ähnlichen Verbindungen erfolgte keine Induktion. Für EtfA und EtfB wurde eine Funktion als universeller Elektronenüberträger des Phenylacrylat-Reduktionssystems in A. woodii postuliert. Für die H2-abhängige Reduktion von Caffeat und anderer Phenylacrylate wurde die folgende Reaktionssequenz postuliert: die Oxidation des Elektronendonors H2 durch eine Hydrogenase geht einher mit der Bildung von reduziertem Ferredoxin. Dieses wird durch eine membranständige Ferredoxin-NAD+-Oxidoreduktase oxidiert, die den Transfer der Elektronen auf NAD+ mit der Translokation von Na+ koppelt. Ein aus EtfA und EtfB gebildetes ETFP fungiert dann als Elektronenüberträger zwischen NADH + H+ und einer löslichen Phenylacrylatreduktase.Acetobacterium woodii was grown with fructose in the presence of caffeate as electron acceptor. Cells were treated with lysozyme and broken up applying low preassure by passage through a „French Press“ cell yielding cell-free extract. All manipulations were performed under strictly anaerobic conditions. This cell-free extract mediated H2-dependent caffeate reduction with rates between 8.7 and 18.7 nmol/min x mg protein. The activity was strictly dependent on the presence of ATP. After separating cell-free extract into the cytoplasmic and the membraneous fraction H2-dependent caffeate reduction was exclusively located in the cytoplasm. The addition of membranes lead to no stimulation of the activity. No activity was found in the membraneous fraction. Various compounds were tested to act as artficial electron donors for caffeate reduction. Tetramethyl phenylendiamine, 1,5-diphenylcarbazide and reduced methyl viologen did not mediate caffeate reduction. In the presence of phenylenediamine caffeate reduction was observed in cell-free extract and the cytoplasmic fraction. In the membraneous fraction no reduction of caffeate was observed with phenylenediamine serving as electron donor. NADH acted as a physiological electron donor for the reduction of caffeate. NADH-dependent caffeate reduction was exclusively located in the cytoplasmic fraction and was strictly dependent on the presence of ATP. The observation that NADH acts as an electron donor for caffeate reduction raised the question how NADH is generated during H2-dependent caffeate reduction and which enzymes could be involved. NAD+-dependent hydrogenase activity was identified in the membraneous and the cytoplasmic fraction. About 70% of the activity was located in the soluble fraction. The presence of an electron donor:NAD+-oxidoreductase in A. woodii was investigated. Washed membranes mediated the oxidation of NADH with ferricyanide or benzyl viologen serving as electron acceptors. This observation was taken as a hint for the presence of a NAD+-reductase, since these enzymes are usually reversible. A hydrogenase could be excluded to mediate this activity since the NADH-oxidizing activity was insensitive towards oxygen. The activity of the NADH-oxidizing enzyme could not be stimulated by Na+. In analogy to Na+-translocating NADH:quinone-reductases the NADH-oxidizing activity located at washed membranes was completly inhibited by micro-molar concentrations of Ag+ or Cu2+. Washed membranes of A. woodii mediated the reduction of NAD+ with reduced ferredoxin serving as electron donor. It could not be clarified, whether this activity was mediated by the membrane-bound NADH-oxidizing enzyme. Ferredoxin-dependent NAD+-reduction was completely inhibited by micro-molar concentrations of Ag+. This inihibition was most likely unspecific. Using 2D gel electrophoresis two caffeate-induced proteins were identified that were subsequently subjected to ESI-MS/MS. A comparision of the obtained peptide sequences with protein sequences from data bases revealed great identities to the large alpha-subunit and the small beta-subunit respectively of heterodimeric electron transfer flavorproteins (ETFP). The caffeate-induced proteins were designated EtfA and EtfB respectively. Based on the peptide sequences degenerated oligonucleotides were deduced and gene fragements coding for EtfA and EtfB were amplified. Analysis of the amino acid sequences deduced from the PCR products confirmed that EtfA and EtfB are subunits of an ETFP. The ability to reduce caffeate is not a constitutive feature of A. woodii but is induced in the presence of the phenyl acrylate. The specificity of this induction was investigated. Suspensions of resting cells prepared from caffeate-induced cells reduced besides caffeate other phenyl acrylates like ferulate or p-coumarate with H2 serving as electron donor. This observation implies that the reduction of phenyl acrylates in A. woodii is accomplished by one single induceable enzymatic system. EtfA and EtfB were heterologously overproduced as MalE-fusion proteins and were purified. Antibodies were generated against MalE-EtfA and MalE-EtfB. Western blot analysis revealed that the production of EtfA and EtfB is induced by various phenyl acrylates. This induction was independent of the growth substrate. In the presence of compounds structurally similar to phenyl acrylates no induction was observed. Thus, it is postulated that EtfA/EtfB act as the universal electron mediator of the phenyl acrylate reducing system in A. woodii. The following reaction sequence was postulated for the reduction of caffeate and other phenyl acrylates: the oxidation of the electron donor H2 is accompanied by the generation of reduced ferredoxin. The latter is oxidized by a membrane-bound ferredoxin-NAD+-oxidoreductase that couples electron transfer to NAD+ with the translocation of Na+. An ETFP consisting of EtfA and EtfB mediates the electron transfer between NADH and a soluble phenyl acrylate reductase

Разработка схемы электроснабжения Вахского нефтяного месторождения ОАО «Томскнефть» ВНК

Выпускная квалификационная работа 185 с., 49 рис., 60 табл., 29 источника, 10 приложение. Ключевые слова: расчётная нагрузка, электроснабжение, выбор сечения, защитной аппаратуры. Объектом исследования является нефтяные месторождения ОАО «Томскнефть» ВНК. Цель работы: разработка системы электроснабжения нефтяного месторождения. В процессе исследования произведён поэтапный расчёт нагрузок, выбор сечения линий, оборудования, защитной аппаратуры. В результате исследования спроектирована конкретная модель электроснабжения месторождения. Основные конструктивные, технологические и технико- эксплуатационные характеристики: объект исследования состоит из 5 кустов и прокатно – ремонтного цеха, все относятся II категории по степени надёжности электроснабжения. Область применения: нефтяные месторожAuspuff arbeiten Qualifikation 185 S., 49 Abb., 60 Tab., 29 Quelle, 10 App Stichworte: Gewichtslast, die Energieversorgung, der Schutzausrüstung Gegenstand der Studie ist die ölfelder Publikumsgesellschaft «Tomskneft» Waher ölkonz Ziel der Arbeit: die Entwicklung des Systems der Elektrizitätsversorgung ölfeld Im forschungsprozeß produziert eine schrittweise Berechnung von Lasten, Auswahl der Querschnitt der Leitungen und Geräte, Schutzausrüstung Infolge der Forschung entwickelt ein konkretes Modell der Energieversorgung vorkommen Grundlegende Konstruktive, technologische und betriebstechnische Daten: das Objekt der Forschung besteht aus 5 Sträuchern und rollende – Reparatur-Werkstatt, alle gehören der Kategorie II durch den Grad der Zuverlässigkeit der Stromversorgung Einsatzbereich: ölfe

Alternative evaluation methods for roundness measurements

Requirements to roundness tolerances are a part of the geometrical product specifications. However, the definition for the roundness tolerance according to ISO 1101 considering radial deviations only is not sufficient to assure the functionality of many products. In addition, the form of roundness deviations along the circumference plays a significant rule for rotating machine components. Especially periodic deviations cause vibrations that lead to noise and wear. The Fourier analysis and the corresponding amplitude spectrum deliver information about the properties of the form derived from the magnitude of the different harmonics. This information presents a series of results depending on the harmonics. Therefore, a dedicated tolerance definition in most cases in from of a mathematical equation is used. The currently used tolerance definitions are not standardized and difficult to understand. Often, only one amplitude of the spectrum is significantly larger than the others are and effects functionality. In this case, an algorithm that detects the largest amplitude enables an easier tolerance definition