Development of a high throughput PCR to detect Coxiella burnetii and its application in a diagnostic laboratory over a 7-year period.

Q fever is a worldwide zoonotic infectious disease due to Coxiella burnetii. The clinical presentation may be acute (pneumonia and/or hepatitis) or chronic (most commonly endocarditis). Diagnosis mainly relies on serology and PCR. We therefore developed a quantitative real-time PCR. We first tested blindly its performance on various clinical samples and then, when thoroughly validated, we applied it during a 7-year period for the diagnosis of both acute and persistent C. burnetii infection. Analytical sensitivity (< 10 copies/PCR) was excellent. When tested blindly on 183 samples, the specificity of the PCR was 100% (142/142) and the sensitivity was 71% (29/41). The sensitivity was 88% (7/8) on valvular samples, 69% (20/29) on blood samples and 50% (2/4) on urine samples. This new quantitative PCR was then successfully applied for the diagnosis of acute Q fever and endovascular infection due to C. burnetii, allowing the diagnosis of Q fever in six patients over a 7-year period. During a local small cluster of cases, the PCR was also applied to blood from 1355 blood donors; all were negative confirming the high specificity of this test. In conclusion, we developed a highly specific method with excellent sensitivity, which may be used on sera for the diagnosis of acute Q fever and on various samples such as sera, valvular samples, aortic specimens, bone and liver, for the diagnosis of persistent C. burnetii infection.

Deicer Scaling Resistance of Concrete Mixtures Containing Slag Cement Phase 2: Evaluation of Different Laboratory Scaling Test Methods, TPF-5(100), 2012

With the use of supplementary cementing materials (SCMs) in concrete mixtures, salt scaling tests such as ASTM C672 have been found to be overly aggressive and do correlate well with field scaling performance. The reasons for this are thought to be because at high replacement levels, SCM mixtures can take longer to set and to develop their properties: neither of these factors is taken into account in the standard laboratory finishing and curing procedures. As a result, these variables were studied as well as a modified scaling test, based on the Quebec BNQ scaling test that had shown promise in other research. The experimental research focused on the evaluation of three scaling resistance tests, including the ASTM C672 test with normal curing as well as an accelerated curing regime used by VDOT for ASTM C1202 rapid chloride permeability tests and now included as an option in ASTM C1202. As well, several variations on the proposed draft ASTM WK9367 deicer scaling resistance test, based on the Quebec Ministry of Transportation BNQ test method, were evaluated for concretes containing varying amounts of slag cement. A total of 16 concrete mixtures were studied using both high alkali cement and low alkali cement, Grade 100 slag and Grade 120 slag with 0, 20, 35 and 50 percent slag replacement by mass of total cementing materials. Vinsol resin was used as the primary air entrainer and Micro Air® was used in two replicate mixes for comparison. Based on the results of this study, a draft alternative test method to ASTM C762 is proposed.

An optimized method for establishing high purity murine CD8(+) T cell cultures.

Establishing CD8(+) T cell cultures has been empirical and the published methods have been largely individual laboratory based. In this study, we optimized culturing conditions and show that IL-2 concentration is the most critical factor for the success of establishing CD8(+) T cell cultures. High IL-2 concentration encouraged T cells to non-specifically proliferate, express a B cell marker, B220, and undergo apoptosis. These cells also lose typical irregular T cell morphology and are incapable of sustaining long-term cultures. Using tetramer and intracellular cytokine assessments, we further demonstrated that many antigen-specific T cells have been rendered nonfunctional when expanded under high IL-2 concentration. When IL-2 is used in the correct range, B220-mediated cell depletion greatly enhanced the success rate of such T cell cultures.

An Investigation of the Chemical Method of Determining the Cement Content of Hardened Concrete, MLR-71-01 and R-248, 1971

The Iowa State Highway Commission Laboratory is called upon to determine the cement content of hardened concrete when field problems relating to batch weights are encountered. The standard test for determining the cement content is ASTM C-85. An investigation of this method by the New Jersey State Highway Department involving duplicate samples and four cooperating laboratories produced very erratic results, however, the results obtained by this method have not been directly compared to known cement contents of concrete made with various cements and various aggregates used in Iowa.

Evaluation of Long-Term Field Performance of Cold In-Place Recycled Roads: Field and Laboratory Testing, TR-502, 2007

Cold in-place recycling (CIR) has become an attractive method for rehabilitating asphalt roads that have good subgrade support and are suffering distress related to non-structural aging and cracking of the pavement layer. Although CIR is widely used, its use could be expanded if its performance were more predictable. Transportation officials have observed roads that were recycled under similar circumstances perform very differently for no clear reason. Moreover, a rational mix design has not yet been developed, design assumptions regarding the structural support of the CIR layer remain empirical and conservative, and there is no clear understanding of the cause-effect relationships between the choices made during the design/construction process and the resulting performance. The objective of this project is to investigate these relationships, especially concerning the age of the recycled pavement, cumulative traffic volume, support conditions, aged engineering properties of the CIR materials, and road performance. Twenty-four CIR asphalt roads constructed in Iowa from 1986 to 2004 were studied: 18 were selected from a sample of roads studied in a previous research project (HR-392), and 6 were selected from newer CIR projects constructed after 1999. This report describes the results of comprehensive field and laboratory testing for these CIR asphalt roads. The results indicate that the modulus of the CIR layer and the air voids of the CIR asphalt binder were the most important factors affecting CIR pavement performance for high-traffic roads. For low-traffic roads, the wet indirect tensile strength significantly affected pavement performance. The results of this research can help identify changes that should be made with regard to design, material selection, and construction in order to improve the performance and cost-effectiveness of future recycled roads.

A new method to simulate the hydrological state of soil under natural conditions

Micro, macro and mesofauna in the soil often respond to fluctuating environmental conditions, resulting in changes of abundance and community structure. Effects of changing soil parameters are normally determined with samples taken in the field and brought to the laboratory, i.e. where natural environmental conditions may not apply. We devised a method (STAFD - soil tubes for artificial flood and drought), which simulates the hydrological state of soil in situ using implanted cores. Control tubes were compared with treatment tubes in which floods of 15, 30, 60 and 90 days, and droughts of 60, 90 and 120 days were simulated in the field. Flooding and drought were found to reduce number of individuals in all soil faunal groups, but the response to drought was slower and not in proportion to the expected decrease of the water content. The results of the simulated floods in particular show the value of the STAFD method for the investigation of such extreme events in natural habitats.

A radiochemical method for carbamoyl-phosphate synthetase I: application to rats fed a hyperproteic diet

A method for the measurement of carbamoyl-phosphate synthetase I activity in animal tissues has been developed using the livers of rats under normal and hyperproteic diets. The method is based on the incorporation of 14C-ammonium bicarbonate to carbamoyl-phosphate in the presence of ATP-Mg and N-acetyl-glutamate. The reaction is stopped by chilling, lowering the pH and adding ethanol. Excess bicarbonate is flushed out under a gentle stream of cold CO2. The only label remaining in the medium was that incorporated into carbamoyl-phosphate, since all 14C-CO2 from bicarbonate was eliminated. The method is rapid and requires only a low pressure supply of CO2 to remove the excess substrate. The reaction is linear up to 10 min using homogenate dilutions of 1:20 to 1:200 (w/v). Rat liver activity was in the range of 89±8 nkat/g. Hyperproteic diet resulted in a significant 1.4-fold increase. The design of the method allows for the processing of multiple samples at the same time, and incubation medium manipulation is unnecessary, since the plastic incubation vial and its contents are finally counted together.

Vitamin B12 Deficiency in the aged: Laboratory Diagnosis, Prevalence and Clinical Profile

B₁₂-vitamiinin puute iäkkäillä: laboratoriodiagnostiikka, yleisyys ja yhteys sairastavuuteen Tausta: B12-vitamiinin puute on yleistä iäkkäillä ja se tulisi todeta riittävän varhaisessa vaiheessa palautumattomien vaurioiden estämiseksi. On epäselvää pitäisikö diagnostiikka kohdistaa tiettyihin riskiryhmiin vai mahdollisesti seuloa valikoimatonta vanhusväestöä. Myöskään yksimielisyyttä laboratoriotutkimusten valinnasta ei ole. Tavoitteet: Tutkimuksen tarkoituksena oli evaluoida uutta HoloTC RIA menetelmää ja tuottaa viitearvot sille, selvittää B12-vitamiinin puutteen yleisyys, yhteys sairastavuuteen ja mahdolliset riskitekijät suomalaisessa vanhusväestössä, arvioida munuaisfunktion vaikusta B12-vitamiinin puutteen laboratoriotutkimuksiin ja näiden perusteella ehdottaa suomalaiseen terveydenhuoltoon sopivaa laboratoriotutkimusstrategiaa. Aineisto ja menetelmät: Liedon iäkkäät -tutkimuksen vanhusaineisto on edustava otos yhden kunnan yli 65-vuotiaasta väestöstä, yhteensä 1260 henkilöä. Tutkittavat kävivät lääkärintarkastuksessa, ja heistä on käytettävissä runsaasti laboratoriotutkimuksia sekä tiedot sairauksista, ruokavaliosta, lääkkeiden ja vitamiinivalmisteiden käytöstä, dementiaseula ja depressiokysely. Viitearvoaineistoa varten kerättiin näytteet 84 vapaaehtoisesta terveestä aikuisesta ja menetelmäevaluaatiota varten 107 sairaalapotilaasta. Tulokset: HoloTC RIA menetelmän toistettavuus oli hyvä manuaalimenetelmäksi. 95%:n viiteväli holotranskobalamiinille oli 37-171 pmol/l. Kaikilla tutkittavilla, joilla oli muilla laboratoriotutkimuksilla osoitettu todennäköinen B12-vitamiinin puute, myös holotranskobalamiini oli viitealueen alarajaa pienempi. Suurentuneella kystatiini C-pitoisuudella osoitettu munuaisten vajaatoiminta korreloi voimakkaasti homokysteiinin (rs=0.53, p<0.001) ja metyylimalonihapon (rs=0.27, p<0.001) pitoisuuksiin, mutta ei kokonais-B12-vitamiinin (rs=- 0.04, p=0.227) tai holotranskobamiinin (rs=-0.01, p=0.817) pitoisuuksiin. Suomalaisessa vanhusväestössä B12-vitamiinin puutteen prevalenssi oli 12%. Kokonais- B12-vitamiinin pitoisuus oli matala (<150 pmol/l) 6%:lla. Miessukupuoli (OR 1.9, 95% CI 1.2-2.9), ikä ≥75 (OR 2.2, 95% CI 1.4-3.4) ja maitotuotteiden välttäminen (OR 2.3, 95% CI 1.2-4.4) lisäsivät B12-vitamiinin puutteen riskiä, mutta anemia (OR 1.3, 95% CI 0.7-2.3) tai makrosytoosi (OR 1.2, 95% CI 0.6-2.7) eivät. Päätelmät: Diagnosoimaton B12-vitamiinin puute on yleistä iäkkäillä, mutta kliinisesti merkityksellistä spesifistä riskiryhmää ei löydy. Koska anemian ja makrosytoosin puuttuminen ei poissulje B12-vitamiinin puutetta ja munuaisten vajaatoiminta heikentää metabolisten merkkiaineiden käyttökelpoisuutta, kokonais-B12-vitamiinia suositellaan ensisijaiseksi laboratoriotutkimukseksi epäiltäessä B12-vitamiinin puutetta ja tarvittaessa varmentavina tutkimuksina käytetään homokysteiiniä ja holotranskobalamiinia.

Effective notch method assessment of misalignment for fatigue loaded welds

 The objective of this thesis work was to assess axial misalignment in fatigue loaded welds using the effective notch method. As a result, the fatigue behaviour of non-load carrying cruciform fillet welded joint under cyclic tensile loading has been studied. Various degrees of axial misalignment have been found in one series of non-load carrying cruciform fillet welded joints used in a laboratory investigation. As a result, it was important to carry out a comprehensive investigation since axial misalignment forms part of thequality of fatigue loaded structure and can reduce the fatigue strength. To extend the study, the correlation between fatigue strength and stress ratio, as well as stress concentration factor, were also studied. Moreover, a closer investigation of place of crack initiation and its dependence on weld sequence and imperfections of test specimen (angular distortion) was studied. For the fatigue class calculations, FEM (finite element method) and the effectivenotch approach are used. The addressed variable is the axial misalignment whichis introduce by modeling the entire joint. Fracture mechanics based calculations are also used and quantitatively compared with effective notch and experimental results.

Development of a Testing Method for Convertibility of Paperboard

Työssä tutkittiin muovattujen kartonkivuokien sekä muovattujen kartonkinäytteiden rinnastettavuutta. Puristusvaiheen prosessiolosuhteiden miellettiin vaikuttavan eniten multidimensionaliseen muodonmuutokseen. Multidimensionaalista muodonmuutosta simuloitiin uudella muovaamiseen soveltuvalla muovauslaitteella. Kirjallisuusosassa keskeisiä teemoja ovat kartongin muovaus sekä kuitupohjaisen materiaalin reologinen käyttäytyminen. Kirjallisuusosassa esitellään lisäksi yksi tekninen sovellus, jonka avulla kyetään ennustamaan kuitumateriaalin muovautuvuutta sekä mittaamaan tapahtunutta muodonmuutosta. Prosessiparametrien teoreettista vaikutustakuituihin tarkastellaan myös kirjallisuusosassa. Kokeellisessa osassa toteutettiin kartonkivuokien valmistus puristamalla. Vastaavilla prosessiparametreilla muovattiin myös pienemmät testinäytteet. Perinteiset yksidimensionaliset deformaatiomittaukset toteutettiin lujuusominaisuuksien laboratoriomäärityksinä. Myös kitka, joka toimii tärkeänä muuttujana prässäysprosessissa, mitattiin laboratorio-olosuhteissa. Tämän työn tulokset osoittavat uuden kehitetyn muovausmenetelmän toimivuuden. Asema-voima kuvaajat ovat selkeitä sekä helposti luettavia. Tuloksissa havaittiin materiaalin muovauspotentiaalin sekä asema-voima kuvaajan välillä vallitseva yhteys. Erittäin merkittävä huomio oli myös, että muovipäällystetyllä kartongilla oli yhteys päällystämättömän kartongin asema-voima kuvaajaan. Tämä tulos osoittaa, että muovipäällystetyn kartongin muovautuvuutta voi olla mahdollista ennustaa pohjakartongin muovautuvuustulosten perusteella. Perinteiset yksidimensionaliset laboratoriomittaukset eivät kykene antamaan riittävää informaatiota muovautuvuuden ennustamiseen. Tästä näkökulmasta on tärkeää että kartongin multidimensionalista muotoutuvuutta voidaankin tutkia kehitetyllä muovausmenetelmällä.

Percutaneous biopsy through the foramen ovale for parasellar lesions: surgical anatomy, method, and indications.

Knowledge of the pathological diagnosis before deciding the best strategy for treating parasellar lesions is of prime importance, due to the relative high morbidity and side-effects of open direct approaches to this region, known to be rich in important vasculo-nervous structures. When imaging is not evocative enough to ascertain an accurate pathological diagnosis, a percutaneous biopsy through the transjugal-transoval route (of Hartel) may be performed to guide the therapeutic decision.The chapter is based on the authors' experience in 50 patients who underwent the procedure over the ten past years. There was no mortality and only little (mostly transient) morbidity. Pathological diagnosis accuracy of the method revealed good, with a sensitivity of 0.83 and a specificity of 1.In the chapter the authors first recall the surgical anatomy background from personal laboratory dissections. They then describe the technical procedure, as well as the tissue harvesting method. Finally they define indications together with the decision-making process.Due to the constraint trajectory of the biopsy needle inserted through the Foramen Ovale, accessible lesions are only those located in the Meckel trigeminal Cave, the posterior sector of the cavernous sinus compartment, and the upper part of the petroclival region.The authors advise to perform this percutaneous biopsy method when imaging does not provide sufficient evidence of the pathological nature of the lesion, for therapeutic decision. Goal is to avoid unnecessary open surgery or radiosurgery, also inappropriate chemo-/radio-therapy.

A radiochemical method for carbamoyl-phosphate synthetase I: application to rats fed a hyperproteic diet

A method for the measurement of carbamoyl-phosphate synthetase I activity in animal tissues has been developed using the livers of rats under normal and hyperproteic diets. The method is based on the incorporation of 14C-ammonium bicarbonate to carbamoyl-phosphate in the presence of ATP-Mg and N-acetyl-glutamate. The reaction is stopped by chilling, lowering the pH and adding ethanol. Excess bicarbonate is flushed out under a gentle stream of cold CO2. The only label remaining in the medium was that incorporated into carbamoyl-phosphate, since all 14C-CO2 from bicarbonate was eliminated. The method is rapid and requires only a low pressure supply of CO2 to remove the excess substrate. The reaction is linear up to 10 min using homogenate dilutions of 1:20 to 1:200 (w/v). Rat liver activity was in the range of 89±8 nkat/g. Hyperproteic diet resulted in a significant 1.4-fold increase. The design of the method allows for the processing of multiple samples at the same time, and incubation medium manipulation is unnecessary, since the plastic incubation vial and its contents are finally counted together.

Jevons, Mill and the Private Laboratory of the Mind

In this paper the rift between Jevons and Mill over the method of political economy serves as a prehistory to recent attempts of behavioural economists to once again explain economic behaviour by taking recourse to mankind's physiology. While Mill relied on the association psychology and its introspective method to establish indubitable first principles, Jevons scorned all recourse to introspection. As exemplified for Jevons's theory of labour, psychophysiology gave Jevons the means to think about economic behaviour in terms of functional form, and promised its assessment by means of experiments. Thus levelling down the Victorian distinction between mind and matter, Jevons turned political economy into social physics.

Identification of new transformation products during enzymatic treatment of tetracycline and erythromycin antibiotics at laboratory scale by an on-line turbulent flow liquid-chromatography coupled to a high resolution mass spectrometer LTQ-Orbitrap

This work describes the formation of transformation products (TPs) by the enzymatic degradation at laboratory scale of two highly consumed antibiotics: tetracycline (Tc) and erythromycin (ERY). The analysis of the samples was carried out by a fast and simple method based on the novel configuration of the on-line turbulent flow system coupled to a hybrid linear ion trap – high resolution mass spectrometer. The method was optimized and validated for the complete analysis of ERY, Tc and their transformation products within 10 min without any other sample manipulation. Furthermore, the applicability of the on-line procedure was evaluated for 25 additional antibiotics, covering a wide range of chemical classes in different environmental waters with satisfactory quality parameters. Degradation rates obtained for Tc by laccase enzyme and ERY by EreB esterase enzyme without the presence of mediators were ∼78% and ∼50%, respectively. Concerning the identification of TPs, three suspected compounds for Tc and five of ERY have been proposed. In the case of Tc, the tentative molecular formulas with errors mass within 2 ppm have been based on the hypothesis of dehydroxylation, (bi)demethylation and oxidation of the rings A and C as major reactions. In contrast, the major TP detected for ERY has been identified as the “dehydration ERY-A”, with the same molecular formula of its parent compound. In addition, the evaluation of the antibiotic activity of the samples along the enzymatic treatments showed a decrease around 100% in both cases

A study of combustion phenomena in circulating fluidized beds by developing and applying experimental and modelling methods for laboratory-scale reactors

The results shown in this thesis are based on selected publications of the 2000s decade. The work was carried out in several national and EC funded public research projects and in close cooperation with industrial partners. The main objective of the thesis was to study and quantify the most important phenomena of circulating fluidized bed combustors by developing and applying proper experimental and modelling methods using laboratory scale equipments. An understanding of the phenomena plays an essential role in the development of combustion and emission performance, and the availability and controls of CFB boilers. Experimental procedures to study fuel combustion behaviour under CFB conditions are presented in the thesis. Steady state and dynamic measurements under well controlled conditions were carried out to produce the data needed for the development of high efficiency, utility scale CFB technology. The importance of combustion control and furnace dynamics is emphasized when CFB boilers are scaled up with a once through steam cycle. Qualitative information on fuel combustion characteristics was obtained directly by comparing flue gas oxygen responses during the impulse change experiments with fuel feed. A one-dimensional, time dependent model was developed to analyse the measurement data Emission formation was studied combined with fuel combustion behaviour. Correlations were developed for NO, N2O, CO and char loading, as a function of temperature and oxygen concentration in the bed area. An online method to characterize char loading under CFB conditions was developed and validated with the pilot scale CFB tests. Finally, a new method to control air and fuel feeds in CFB combustion was introduced. The method is based on models and an analysis of the fluctuation of the flue gas oxygen concentration. The effect of high oxygen concentrations on fuel combustion behaviour was also studied to evaluate the potential of CFB boilers to apply oxygenfiring technology to CCS. In future studies, it will be necessary to go through the whole scale up chain from laboratory phenomena devices through pilot scale test rigs to large scale, commercial boilers in order to validate the applicability and scalability of the, results. This thesis shows the chain between the laboratory scale phenomena test rig (bench scale) and the CFB process test rig (pilot). CFB technology has been scaled up successfully from an industrial scale to a utility scale during the last decade. The work shown in the thesis, for its part, has supported the development by producing new detailed information on combustion under CFB conditions.