Comparison of the developmental effects of two mercury compounds on glial cells and neurons in aggregate cultures of rat telencephalon.

A three-dimensional cell culture system was used as a model to study the influence of low levels of mercury in the developing brain. Aggregating cell cultures of fetal rat telencephalon were treated for 10 days either during an early developmental period (i.e., between days 5 and 15 in vitro) or during a phase of advanced maturation (i.e., between days 25 and 35) with mercury. An inorganic (HgCl2) and an organic mercury compound (monomethylmercury chloride, MeHgCl) were examined. By monitoring changes in cell type-specific enzymes activities, the concentration-dependent toxicity of the compounds was determined. In immature cultures, a general cytotoxicity was observed at 10(-6) M for both mercury compounds. In these cultures, HgCl2 appeared somewhat more toxic than MeHgCl. However, no appreciable demethylation of MeHgCl could be detected, indicating similar toxic potencies for both mercury compounds. In highly differentiated cultures, by contrast, MeHgCl exhibited a higher toxic potency than HgCl2. In addition, at 10(-6) M, MeHgCl showed pronounced neuron-specific toxicity. Below the cytotoxic concentrations, distinct glia-specific reactions could be observed with both mercury compounds. An increase in the immunoreactivity for glial fibrillary acidic protein, typical for gliosis, could be observed at concentrations between 10(-9) M and 10(-7) M in immature cultures, and between 10(-8) M and 3 x 10(-5) M in highly differentiated cultures. A conspicuous increase in the number and clustering of GSI-B4 lectin-binding cells, indicating a microglial response, was found at concentrations between 10(-10) M and 10(-7) M. These development-dependent and cell type-specific effects may reflect the pathogenic potential of long-term exposure to subclinical doses of mercury.

High-Resolution Mass Spectrometry applied to the identification of new metabolites and transformation products of quinolones in chicken muscle tissues

The aim of this work was the identification of new metabolites and transformation products (TPs) in chicken muscle from Enrofloxacin (ENR), Ciprofloxacin (CIP), Difloxacin (DIF) and Sarafloxacin (SAR), which are antibiotics that belong to the fluoroquinolones family. The stability of ENR, CIP, DIF and SAR standard solutions versus pH degradation process (from pH 1.5 to 8.0, simulating the pH since the drug is administered until its excretion) and freeze-thawing (F/T) cycles was tested. In addition, chicken muscle samples from medicated animals with ENR were analyzed in order to identify new metabolites and TPs. The identification of the different metabolites and TPs was accomplished by comparison of mass spectral data from samples and blanks, using liquid chromatography coupled to quadrupole time-of-flight (LC-QqToF) and Multiple Mass Defect Filter (MMDF) technique as a pre-filter to remove most of the background noise and endogenous components. Confirmation and structure elucidation was performed by liquid chromatography coupled to linear ion trap quadrupole Orbitrap (LC-LTQ-Orbitrap), due to its mass accuracy and MS/MS capacity for elemental composition determination. As a result, 21 TPs from ENR, 6 TPs from CIP, 14 TPs from DIF and 12 TPs from SAR were identified due to the pH shock and F/T cycles. On the other hand, 14 metabolites were identified from the medicated chicken muscle samples. Formation of CIP and SAR, from ENR and DIF, respectively, and the formation of desethylene-quinolone were the most remarkable identified compounds.

A Study of the Frictional Properties of New Asphaltic Concrete Pavements in Iowa, MLR-78-03, 1978

Seven asphaltic concrete resurfacing projects were tested for their frictional properties to determine the age-friction relationship of new paving. Projects studied included Type A asphaltic concrete which is generally used for higher traffic volume roads and Type B asphaltic concrete, a lower type material. Also included in the study were asphaltic concretes containing Type 3 and Type 4 coarse aggregate texture classifications. The classifications are based upon material type and grain size composition. Surfaces both with and without sprinkle treatment aggregates were also included. The data gathered suggests that properly designed and placed dense graded asphaltic concrete mixes are adequate to serve the traveling public at all ages tested.

Maintenance and Design of Steel Abutment Piles in Iowa Bridges, TR-622, 2014

Soil consolidation and erosion caused by roadway runoff have exposed the upper portions of steel piles at the abutments of numerous bridges, leaving them susceptible to accelerated corrosion rates due to the abundance of moisture, oxygen, and chlorides at these locations. This problem is compounded by the relative inaccessibility of abutment piles for close-up inspection and repair. The objective of this study was to provide bridge owners with recommendations for effective methods of addressing corrosion of steel abutment piles in existing and future bridges A review of available literature on the performance and protection of steel piles exposed to a variety of environments was performed. Eight potential coating systems for use in protecting existing and/or new piles were selected and subjected to accelerated corrosion conditions in the laboratory. Two surface preparation methods were evaluated in the field and three coating systems were installed on three piles at an existing bridge where abutment piles had been exposed by erosion. In addition, a passive cathodic protection (CP) system using sacrificial zinc anodes was tested in the laboratory. Several trial flowable mortar mixes were evaluated for use in conjunction with the CP system. For existing abutment piles, application of a protective coating system is a promising method of mitigating corrosion. Based on its excellent performance in accelerated corrosion conditions in the laboratory on steel test specimens with SSPC-SP3, -SP6, and -SP10 surface preparations, glass flake polyester is recommended for use on existing piles. An alternative is epoxy over organic zinc rich primer. Surface preparation of existing piles should include abrasive blast cleaning to SSPC-SP6. Although additional field testing is needed, based on the results of the laboratory testing, a passive CP system could provide an effective means of protecting piles in existing bridges when combined with a pumped mortar used to fill voids between the abutment footing and soil. The addition of a corrosion inhibitor to the mortar appears to be beneficial. For new construction, shop application of thermally sprayed aluminum or glass flake polyester to the upper portion of the piles is recommended.

Field performance of new cowpea cultivars inoculated with efficient nitrogen-fixing rhizobial strains in the Brazilian Semiarid

The objective of this work was to evaluate the contribution of efficient nitrogen-fixing rhizobial strains to grain yield of new cowpea cultivars, indicated for cultivation in the Brazilian Semiarid region, in the sub-medium of the São Francisco River Valley. Two experiments were set up at the irrigated perimeters of Mandacaru (Juazeiro, state of Bahia) and Bebedouro (Petrolina, state of Pernambuco). The treatments consisted of single inoculation of five rhizobial strains - BR 3267, BR 3262, INPA 03-11B, UFLA 03-84 (Bradyrhizobiumsp.), and BR 3299T(Microvirga vignae) -, besides a treatment with nitrogen and a control without inoculation or N application. The following cowpea cultivars were evaluated: BRS Pujante, BRS Tapaihum, BRS Carijó, and BRS Acauã. A randomized complete block design, with four replicates, was used. Inoculated plants showed similar grain yield to the one observed with plants fertilized with 80 kg ha-1 N. The cultivars BRS Tapaihum and BRS Pujante stood out in grain yield and protein contents when inoculated, showing their potential for cultivation in the sub-medium of the São Francisco River Valley.

Design of adjustable packaging line

Diplomityön tavoite oli tutkia, mitä toimintoja ja tekniikoita uusi, joustava kartonkipakkauslinja sisältää ja mitkä suuntaukset pakkausteollisuudessa tulevat olemaan tärkeitä tulevaisuudessa. Pakkauslinjan päätoimintoja tarkasteltiin nykyisten jatulevaisuuden tekniikoiden pohjalta. Erityisesti tarkasteltiin laser-sovellusten käytön mahdollisuutta pakkauslinjan eri osatoiminnoissa. Katsaus pakkausteollisuuden tulevaisuuden näkymiin luotiin kirjallisuuden ja aikaisempien tutkimusten pohjalta, minkä perusteella työssä oletetaan, että yksilölliset ja monitoimipakkaukset tulevat lisääntymään tulevaisuudessa. Eri tuotantoerien välillä olevat asetusajat tulee saada minimoitua, mutta millä keinoin joustavuus on saavutettavissa? Yksi ratkaisu pakkausten valmistamisessa on käyttää robottisolua, mikä on mahdollista luultavasti ainakin kuppimuotoisten pakkausten kohdalla. Muutenkin robotiikka on lisääntymässä pakkausteollisuudessa. Digitaalisten painotekniikoiden kehitys on mahdollistanut yksilölliset painatukset. Tulevaisuudessa painatus on mahdollista tehdä pakkauslinjan loppupäässä, jopa vasta täytön ja suljennan jälkeen. Laserleikkaus on jo nyt käytössä, mutta tulevaisuudessa myös saumaus ja perinteinen nuuttaus on mahdollista tehdä lasersovelluksia käyttäen. Kehittynyt, väyläpohjainen ohjausjärjestelmä on tulevaisuudessa välttämätön joustavassa pakkauslinjassa. Internetin välityksellä toimiva etäohjattu virheenkorjausdiagnostiikka tulee myös olemaan itsestäänselvyys tulevaisuudessa. Kustannussäästöjä voidaan saavuttaa käyttämälläpakkauslinjassa modulaarista rakennetta. Standardiosien ja standardiosajärjestelmien käyttäminen pienentää myös käyttö- ja huoltokustannuksia. Tärkeää on kuitenkin muistaa, ettei joustavuutta voida saavuttaa pelkästään yhtä ominaisuutta tai tekniikkaa hyödyntäen vaan monia menetelmiä yhdistäen. Suunniteltavan pakkauslinjan toiminta on myös hyvä varmistaa käyttäen apuna mallinnusta ja simulointia.

Design of an Embedded Computer Board for Networking Applications

Tämä diplomityö tehtiin Convergens Oy:lle. Convergens on elektroniikan suunnittelutoimisto, joka on erikoistunut sulautettuihin järjestelmiin sekä tietoliikennetekniikkaan. Diplomityön tavoitteena oli suunnitella tietokonekortti tietoliikennesovelluksia varten asiakkaalle, jolta vaatimusmäärittelyt tulivat. Työ on rajattu koskemaan laitteen prototyypin suunnittelua. Työssä suunnitellaan pääasiassa WLAN-tukiaseman tietokone. Tukiasema onasennettavissa toimistoihin, varastoihin, kauppoihin sekä myös liikkuvaan ajoneuvoon. Suunnittelussa on otettu nämä asiat huomioon, ja laitteen akun pystyy lataamaan muun muassa auton akulla. Langattomat tekniikat ovat voimakkaasti yleistymässä, ja tämän työn tukiasema tarjoaakin varteenotettavan vaihtoehdon lukuisilla ominaisuuksillaan. Mukana on mm. GPS, Bluetooth sekä Ethernet-valmius. Langattomien tekniikoiden lisäksi myös sulautetut järjestelmät ovat voimakkaasti yleistymässä, ja nykyään mikroprosessoreita löytääkin lähesmistä vain. Tässä projektissa käytetty prosessori on nopeutensa puolesta kilpailukykyinen, ja siitä löytyy useita eri rajapintoja. Jatkossa tietokonekortille on myös tulossa WiMAX-tuki, joka lisää tukiaseman tulevaisuuden arvoa asiakkaalle. Projektiin valittu Freescalen MPC8321E-prosessori on PowerPC-arkkitehtuuriin perustuva ja juuri markkinoille ilmestynyt. Tämä toi mukanaan lisähaasteen, sillä kyseisestä prosessorista ei ollut vielä kaikkea tietoa saatavilla. Mekaniikka toi omat haasteensa mukanaan, sillä se rajoitti piirilevyn koonniin, että ylimääräistä piirilevytilaa ei juurikaan jäänyt. Tämän takia esimerkiksi DDR-muistit olivat haastavia reitittää, sillä muistivetojen on oltava melko samanpituisia keskenään. Käyttöjärjestelmänä projektissa käytetään Linuxia. Suunnittelu alkoi keväällä 2007 ja toimiva prototyyppi oli valmis alkusyksystä. Prototyypin testaus osoitti, että tietokonekortti kykenee täyttämään kaikki asiakkaan vaatimukset. Prototyypin testauksessa löytyneet viat ja optimoinnit on tarkoitus korjata tuotantomalliin, joten se antaa hyvän pohjan jatkosuunnittelua varten.

Structure-Based, Rational Design of T Cell Receptors.

Adoptive cell transfer using engineered T cells is emerging as a promising treatment for metastatic melanoma. Such an approach allows one to introduce T cell receptor (TCR) modifications that, while maintaining the specificity for the targeted antigen, can enhance the binding and kinetic parameters for the interaction with peptides (p) bound to major histocompatibility complexes (MHC). Using the well-characterized 2C TCR/SIYR/H-2K(b) structure as a model system, we demonstrated that a binding free energy decomposition based on the MM-GBSA approach provides a detailed and reliable description of the TCR/pMHC interactions at the structural and thermodynamic levels. Starting from this result, we developed a new structure-based approach, to rationally design new TCR sequences, and applied it to the BC1 TCR targeting the HLA-A2 restricted NY-ESO-1157-165 cancer-testis epitope. Fifty-four percent of the designed sequence replacements exhibited improved pMHC binding as compared to the native TCR, with up to 150-fold increase in affinity, while preserving specificity. Genetically engineered CD8(+) T cells expressing these modified TCRs showed an improved functional activity compared to those expressing BC1 TCR. We measured maximum levels of activities for TCRs within the upper limit of natural affinity, K D = ∼1 - 5 μM. Beyond the affinity threshold at K D < 1 μM we observed an attenuation in cellular function, in line with the "half-life" model of T cell activation. Our computer-aided protein-engineering approach requires the 3D-structure of the TCR-pMHC complex of interest, which can be obtained from X-ray crystallography. We have also developed a homology modeling-based approach, TCRep 3D, to obtain accurate structural models of any TCR-pMHC complexes when experimental data is not available. Since the accuracy of the models depends on the prediction of the TCR orientation over pMHC, we have complemented the approach with a simplified rigid method to predict this orientation and successfully assessed it using all non-redundant TCR-pMHC crystal structures available. These methods potentially extend the use of our TCR engineering method to entire TCR repertoires for which no X-ray structure is available. We have also performed a steered molecular dynamics study of the unbinding of the TCR-pMHC complex to get a better understanding of how TCRs interact with pMHCs. This entire rational TCR design pipeline is now being used to produce rationally optimized TCRs for adoptive cell therapies of stage IV melanoma.

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

Solubilization of decane into gemini surfactant with a modified Jeffamine backbone: Design of hierarchical porous silica

Herein, we have investigated the solubilization of decane into a novel nonionic gemini surfactant, myristoyl-end capped Jeffamine, synthesized from a polyoxyalkyleneamine (ED900). Starting from this system, porous silica materials have been prepared. Performing the hydrothermal treatment at low temperature, a slight increase of the mesopore diameter is observed in the presence of decane. Increasing the temperature of the hydrothermal treatment, no swelling effect of decane is detected. By contrast, the pore diameter decreases but better mesopore homogeneity and a larger wall thickness are obtained. At high decane concentration the new myristoyl-end capped Jeffamine/decane/water system forms oil-in-water emulsions, which are used as template for the formation of hierarchical porous silica materials.

On the application of knoevenagel condensation for the synthesis of benzylidene benzothiazine compounds and structural study

The synthesis and physico-chemical properties of new 6-acetylamino or 6-benzoyl-amino 2-benzylidene-4-methyl-4H-benzo[1,4]thiazin-3-ones and 6-benzoylamino or 6-nitro 2-benzylidene-4H-benzo[1,4]thiazin-3-ones are described. These benzylidene benzothiazine compounds were prepared by the Knoevenagel condensation with benzaldehydes. The configurations and conformations of benzylidene benzothiazine derivatives were optimised using the semi-empirical method AM1.

Synthesis and characterization of new amino acyl-4-thiazolidones

A series of heterocyclic compounds with a 4-thiazolidone nucleus and amino acyl moiety were synthesized by protection reaction of thiosemicarbazide using the symmetrical anhydride (Boc)2O and cyclization with chloroacetic acid under mild conditions. Trifluoroacetic acid was used to obtain 4-thiazolidone and the alpha-amino acid condensation reactions were carried out using strategies for peptide synthesis. The characterization of this new class of compounds was performed using IR and ¹H-NMR spectroscopy.

Design of an Event Based Messaging System

B2B document handling is moving from paper to electronic networks and electronic domain very rapidly. Moving, handling and transforming large electronic business documents requires a lot from the systems handling them. This paper explores new technologies such as SOA, event-driven systems and ESB and a scalable, event-driven enterprise service bus is created to demonstrate these new approaches to message handling. As an end result, we have a small but fully functional messaging system with several different components. This is the first larger Java-project done in-house, so on the side we developed our own set of best practices of Java development, setting up configurations, tools, code repositories and class naming and much more.

SYNTHESIS AND BIOLOGICAL ACTIVITY OF NEW SERIES OF ORGANOTIN(IV) ESTERS WITH N,N-DIACETYLGLYCINE

A bioactive N,N-diacetylglycine (NNDAG) and new organotin(IV) complexes (OTCs) (1-7) were synthesized. Spectroscopic techniques were employed to characterize NNDAG and OTCs. FTIR was employed to verify N,N protection of glycine by acetyl groups. The disappearance of υ(OH) at 3000-2600 cm-1 showed de-protonation of free ligand. The Δυ 150<200 cm-1 of OTCs 4-7 verified bidentate coordination with tetrahedral geometry. The Δυ of OTCs1 and 3 was <200 cm-1 exhibitingtrans -octahedral geometry while OTC 2 dimer was assigned a unique sinusoidal view. The 1H NMR spectra of OTCs verified their synthesis by de-protonation of NNDAG and no chemical shift was found downfield for carboxylic acid proton. The 13C, 119Sn NMR and Mass spectrometric data also supported FTIR and 1H NMR descriptions. The OTCs 4, 5, 6 and7 (500 ppm) proved twice as active against Escherichia coli as the standard antibiotic enoxacin (1000 ppm). The promising property of the OTCs (4, 5, 6 and7) is clearly due to their tetrahedral. The OTCs 4and 5 exhibited excellent activity against M. minimum and good activity against T. castaneum.LD50 of all the compounds were determined and OTCs4, 5 and 7 were found to be active.

Simulation of new p-type pixel strip detector with enhanced multiplication effect

This Master’s Thesis is dedicated to the simulation of new p-type pixel strip detector with enhanced multiplication effect. It is done for high-energy physics experiments upgrade such as Super Large Hadron Collider especially for Compact Muon Solenoid particle track silicon detectors. These detectors are used in very harsh radiation environment and should have good radiation hardness. The device engineering technology for developing more radiation hard particle detectors is used for minimizing the radiation degradation. New detector structure with enhanced multiplication effect is proposed in this work. There are studies of electric field and electric charge distribution of conventional and new p-type detector under reverse voltage bias and irradiation. Finally, the dependence of the anode current from the applied cathode reverse voltage bias under irradiation is obtained in this Thesis. For simulation Silvaco Technology Computer Aided Design software was used. Athena was used for creation of doping profiles and device structures and Atlas was used for getting electrical characteristics of the studied devices. The program codes for this software are represented in Appendixes.