Reduction-substitution reactions on [99mTcO4]- with both o-phenylenebis(dimethylarsine) (DIARS) a... more Reduction-substitution reactions on [99mTcO4]- with both o-phenylenebis(dimethylarsine) (DIARS) and various thiols produce a series of monocationic [99Tc(DIARS)2(SR)2]+ complexes. Addition of [99gTcO4]- to the above reaction mixtures allows the characterization of the "carrier-added" complexes by means of reverse-phase high-performance liquid chromatography with radiometric and optical detection systems. The identity of the [99mTc(DIARS)2(SR)2]+ complexes is confirmed by fast atom bombardment mass spectroscopy; equivalence of the [99gTc(DIARS)2-(SR)2]+ and [99mTc(DIARS)2(SR)2]+ species is demonstrated by identical HPLC retention times. All the [99mTc(DIARS)2(SR)2]+ complexes tested accumulate in the myocardium of Sprague-Dawley rats with an average uptake of 1.5-2.0% of injected dose/g at 30 min. Thus, as designed, these nonreducible Tc(III) complexes do not exhibit the rapid myocardial washout observed for reducible Tc(III) complexes. These [99mTc(DIARS)2(SR)2]+ complexes also exhibit an initially high liver uptake, but the presence of ether groups within the thiolate ligands causes this liver uptake to decrease over time without affecting the heart uptake, thereby improving the heart/liver ratio.
An ~se~richiu coli vector to express and purify foreign proteins by fusion to and separation from... more An ~se~richiu coli vector to express and purify foreign proteins by fusion to and separation from maltose-binding protein (Recombinant DNA; malE; factor X, protease; paramyosin; crosslinked amylose; /$galactosidase; Dirofilaria immi~; affinity purification; over expression; phage rl vectors)
The objective of this work was to evaluate extrusion cooking as a means to improve the nutritiona... more The objective of this work was to evaluate extrusion cooking as a means to improve the nutritional properties of Phaseolus vulgaris L. that had been stored either at 42°C and 80% relative humidity for 6 weeks or for periods >1 year in cereal stores in tropical conditions. Storage under these conditions resulted in an increase in cooking time increased (7.7-and 12-fold, respectively) as a result of development of the hard-to-cook (HTC) defect. Single-screw extrusion of the milled beans was carried out at four barrel temperatures and two moisture contents. The extrudate bulk density and water solubility index decreased with increasing temperature, whereas the water absorption index increased due to the higher proportion of gelatinized starch in the extruded samples. Both fresh and HTC beans contained nutritionally significant amounts of lectins, trypsin, and R-amylase inhibitors, which were mostly inactivated by extrusion. Extrusion also caused a considerable redistribution of insoluble dietary fiber to soluble, although the total dietary fiber content was not affected. Changes in solubility involved pectic polysaccharides, arabinose and uronic acids being the main sugars involved. Stored beans subjected to extrusion cooking showed physical and chemical characteristics similar to those of extrudates from fresh beans. SME (kWh kg -1 ) ) screw speed × power (kW) × torque (%) max screw speed × throughput (kg h -1 ) × 100
The exploitation of ''intelligent'' factory set-ups could enhance the competitiveness of the EU t... more The exploitation of ''intelligent'' factory set-ups could enhance the competitiveness of the EU textile and clothing industries, by enabling collaborative design-and-manufacture options, while achieving economies of scope with effective exploitation of (strategic/tactical/execution) flexibility. Simulation turns to be reference aid for developing and acknowledging the appropriate set-ups and the adaptive-schedules. The investigation, besides of reference concepts, summarises a case example, for explanatory purpose. Combined-mode schedules are considered, showing the benefits for prompt returns and leaving open middle/long horizons issues, still with quantitative checks on alternatives provided by virtual reality tests. #
We examine the contribution of Multiple Parton Interactions to Z + n-jets production at the LHC, ... more We examine the contribution of Multiple Parton Interactions to Z + n-jets production at the LHC, n = 2, 3, 4, where the Z boson is assumed to decay leptonically. We compare the results obtained with the correlated GS09 double parton distribution function with those obtained with two instances of fully factorized single parton distribution functions: MSTW2008LO and CTEQ6L1. It appears quite feasible to measure the MPI contribution to Z + 2/3/4 jets already in the first phase of the LHC with a total luminosity of one inverse femtobarn at 7 TeV. If as expected the trigger threshold for single photons is around 80 GeV, Z + 2-jets production may well turn out to be more easily observable than the γ + 3-jets channel. The MPI cross section is dominated by relatively soft events with two jets balancing in transverse momentum.
There are many possibilities for new physics beyond the Standard Model that feature non-standard ... more There are many possibilities for new physics beyond the Standard Model that feature non-standard Higgs sectors. These may introduce new sources of CP violation, and there may be mixing between multiple Higgs bosons or other new scalar bosons. Alternatively, the Higgs may be a composite state, or there may even be no Higgs at all. These non-standard Higgs scenarios have important implications for collider physics as well as for cosmology, and understanding their phenomenology is essential for a full comprehension of electroweak symmetry breaking. This report discusses the most relevant theories which go beyond the Standard Model and its minimal, CP-conserving supersymmetric extension: two-Higgs-doublet models and minimal supersymmetric models with CP violation, supersymmetric models with an extra singlet, models with extra gauge groups or Higgs triplets, Little Higgs models, models in extra dimensions, and models with technicolour or other new strong dynamics. For each of these scenarios, this report presents an introduction to the phenomenology, followed by contributions on more detailed theoretical aspects and studies of possible experimental signatures at the LHC and other colliders.
Journal of Clinical Endocrinology & Metabolism, 2010
Context: Thyroid hormone, requiring adequate maternal iodine intake, is critical for fetal neurod... more Context: Thyroid hormone, requiring adequate maternal iodine intake, is critical for fetal neurodevelopment. Perchlorate decreases thyroidal iodine uptake by competitively inhibiting the sodium/iodide symporter. It is unclear whether environmental perchlorate exposure adversely affects thyroid function in pregnant women. Thiocyanate, derived from foods and cigarette smoke, is a less potent competitive sodium/iodide symporter inhibitor than perchlorate.
Trypanosoma (Nannomonas) congolense comprises morphologically identical but genetically heterogen... more Trypanosoma (Nannomonas) congolense comprises morphologically identical but genetically heterogeneous parasites infective to livestock and other mammalian hosts; three different genotypes of this parasite have been described previously. Restriction enzyme fragment length polymorphisms (RFLPs) in both kinetoplast DNA minicircle and nuclear DNA sequences, and randomly amplified polymorphic deoxyribonucleic acid (RAPD) patterns have been used here to demonstrate the existence of another type of T. (N.) congolense that is genotypically distinct from those that have so far been characterized at the molecular level. A highly repetitive, tandemly arranged DNA sequence and oligonucleotide primers, for use in polymerase chain reaction (PCR) amplification are described, which can be used for specific identification of the trypanosome and its distinction from others within the Nannomonas subgenus.
The transcript encoding a predominant Trypanosoma evansi variable surface glycoprotein RoTat 1.2 ... more The transcript encoding a predominant Trypanosoma evansi variable surface glycoprotein RoTat 1.2 was cloned and expressed as a recombinant protein in Spodoptera frugiperda and Trichoplusia ni insect cells. Its potential as an antigen for specific detection of antibody in serum of dromedary camels affected by surra, was evaluated. In ELISA, the reactivity of the recombinant RoTat 1.2 VSG was similar to that of native RoTat 1.2 VSG. An indirect agglutination reagent was therefore prepared by coupling the recombinant RoTat 1.2 VSG onto latex particles. The performance of the latex agglutination test was evaluated on camel sera, and compared to the performance of CATT/T. evansi and LATEX/T. evansi tests, using the immune trypanolysis assay with T. evansi RoTat 1.2 as a reference test. The relative sensitivity and specificity of the latex coated with recombinant RoTat 1.2 VSG, using a 1:4 serum dilution, were respectively 89.3 and 99.1%. No differences were observed between the performance of latex coated with recombinant RoTat 1.2 VSG and LATEX/T. evansi or CATT/T. evansi. Here, we describe the successful use of the recombinant RoTat 1.2 VSG for detection of specific antibodies induced by T. evansi infections. pH 7.4). Protein A peroxidase (Sigma) was diluted to 0.2 µg/ml in PBS-Tween and incubated for 1 h (150 µl/well). After 5 washes, wells were incubated for 1 h at ambient temperature with 150 µl ABTS substrate-chromogen solution. The latter was prepared from 50 mg ABTS (2,2'-azinobis(3-ethylbenzothiazoline)-6-sulfonic acid, Boehringer, Germany) dissolved in 100 ml of ABTS-buffer (phosphate-citrate-sodium perborate solution, pH 4.6, Boehringer, Germany). The plate was shaken for 10 s and the OD read at 415 nm (Multiskan RC Version 6.0, Labsystems). Corrected OD values were obtained by subtracting the mean OD of the antigen-free control wells from the mean OD of the corresponding antigen containing wells.
Two clones of T. congolenre were derived. one from the other. by growth in mice and administralmn... more Two clones of T. congolenre were derived. one from the other. by growth in mice and administralmn of increasing sub-curat~ve doses of lhe Qpanocldc isometamidiom chloride. One of the clones was Ihus made approunatel! IO more tolerant to lhe bypanocide than the other mP.NA from lhc Iwo clones was used for syntbeas of co~npleu~enta~ DNA (cDNA) An aliquol of the cDNA was convened inlo double-stranded DNA molecules and Qrccl~onall) ligated 8 the EwRIiNotl she of hgt I I. which were subscqucntly espandcd by growih m Ecoli Y IOLX). The libraries obtamed had >I()' independenl phage clones, >90% of which were recombinant. One hundred random recomtxnant clones from each library were subjected to the polymcrase chao reaction (F'CR). using primers flanking rhe nwlliplc clorung site. and were found to contain inserts that vary in size from 0 Skb to 2.3kb.
The transcript encoding a predominant Trypanosoma evansi variable surface glycoprotein RoTat 1.2 ... more The transcript encoding a predominant Trypanosoma evansi variable surface glycoprotein RoTat 1.2 was cloned and expressed as a recombinant protein in Spodoptera frugiperda and Trichoplusia ni insect cells. Its potential as an antigen for specific detection of antibody in serum of dromedary camels affected by surra, was evaluated. In ELISA, the reactivity of the recombinant RoTat 1.2 VSG was similar to that of native RoTat 1.2 VSG. An indirect agglutination reagent was therefore prepared by coupling the recombinant RoTat 1.2 VSG onto latex particles. The performance of the latex agglutination test was evaluated on camel sera, and compared to the performance of CATT/T. evansi and LATEX/T. evansi tests, using the immune trypanolysis assay with T. evansi RoTat 1.2 as a reference test. The relative sensitivity and specificity of the latex coated with recombinant RoTat 1.2 VSG, using a 1:4 serum dilution, were respectively 89.3 and 99.1%. No differences were observed between the performance of latex coated with recombinant RoTat 1.2 VSG and LATEX/T. evansi or CATT/T. evansi. Here, we describe the successful use of the recombinant RoTat 1.2 VSG for detection of specific antibodies induced by T. evansi infections. pH 7.4). Protein A peroxidase (Sigma) was diluted to 0.2 µg/ml in PBS-Tween and incubated for 1 h (150 µl/well). After 5 washes, wells were incubated for 1 h at ambient temperature with 150 µl ABTS substrate-chromogen solution. The latter was prepared from 50 mg ABTS (2,2'-azinobis(3-ethylbenzothiazoline)-6-sulfonic acid, Boehringer, Germany) dissolved in 100 ml of ABTS-buffer (phosphate-citrate-sodium perborate solution, pH 4.6, Boehringer, Germany). The plate was shaken for 10 s and the OD read at 415 nm (Multiskan RC Version 6.0, Labsystems). Corrected OD values were obtained by subtracting the mean OD of the antigen-free control wells from the mean OD of the corresponding antigen containing wells.
Trypanosoma (Nannomonas) congolense comprises morphologically identical but genetically heterogen... more Trypanosoma (Nannomonas) congolense comprises morphologically identical but genetically heterogeneous parasites infective to livestock and other mammalian hosts; three different genotypes of this parasite have been described previously. Restriction enzyme fragment length polymorphisms (RFLPs) in both kinetoplast DNA minicircle and nuclear DNA sequences, and randomly amplified polymorphic deoxyribonucleic acid (RAPD) patterns have been used here to demonstrate the existence of another type of T. (N.) congolense that is genotypically distinct from those that have so far been characterized at the molecular level. A highly repetitive, tandemly arranged DNA sequence and oligonucleotide primers, for use in polymerase chain reaction (PCR) amplification are described, which can be used for specific identification of the trypanosome and its distinction from others within the Nannomonas subgenus.
The transcript encoding a predominant Trypanosoma evansi variable surface glycoprotein RoTat 1.2 ... more The transcript encoding a predominant Trypanosoma evansi variable surface glycoprotein RoTat 1.2 was cloned and expressed as a recombinant protein in Spodoptera frugiperda and Trichoplusia ni insect cells. Its potential as an antigen for specific detection of antibody in serum of dromedary camels affected by surra, was evaluated. In ELISA, the reactivity of the recombinant RoTat 1.2 VSG was similar to that of native RoTat 1.2 VSG. An indirect agglutination reagent was therefore prepared by coupling the recombinant RoTat 1.2 VSG onto latex particles. The performance of the latex agglutination test was evaluated on camel sera, and compared to the performance of CATT/T. evansi and LATEX/T. evansi tests, using the immune trypanolysis assay with T. evansi RoTat 1.2 as a reference test. The relative sensitivity and specificity of the latex coated with recombinant RoTat 1.2 VSG, using a 1:4 serum dilution, were respectively 89.3 and 99.1%. No differences were observed between the performance of latex coated with recombinant RoTat 1.2 VSG and LATEX/T. evansi or CATT/T. evansi. Here, we describe the successful use of the recombinant RoTat 1.2 VSG for detection of specific antibodies induced by T. evansi infections. pH 7.4). Protein A peroxidase (Sigma) was diluted to 0.2 µg/ml in PBS-Tween and incubated for 1 h (150 µl/well). After 5 washes, wells were incubated for 1 h at ambient temperature with 150 µl ABTS substrate-chromogen solution. The latter was prepared from 50 mg ABTS (2,2'-azinobis(3-ethylbenzothiazoline)-6-sulfonic acid, Boehringer, Germany) dissolved in 100 ml of ABTS-buffer (phosphate-citrate-sodium perborate solution, pH 4.6, Boehringer, Germany). The plate was shaken for 10 s and the OD read at 415 nm (Multiskan RC Version 6.0, Labsystems). Corrected OD values were obtained by subtracting the mean OD of the antigen-free control wells from the mean OD of the corresponding antigen containing wells.
Two clones of T. congolenre were derived. one from the other. by growth in mice and administralmn... more Two clones of T. congolenre were derived. one from the other. by growth in mice and administralmn of increasing sub-curat~ve doses of lhe Qpanocldc isometamidiom chloride. One of the clones was Ihus made approunatel! IO more tolerant to lhe bypanocide than the other mP.NA from lhc Iwo clones was used for syntbeas of co~npleu~enta~ DNA (cDNA) An aliquol of the cDNA was convened inlo double-stranded DNA molecules and Qrccl~onall) ligated 8 the EwRIiNotl she of hgt I I. which were subscqucntly espandcd by growih m Ecoli Y IOLX). The libraries obtamed had >I()' independenl phage clones, >90% of which were recombinant. One hundred random recomtxnant clones from each library were subjected to the polymcrase chao reaction (F'CR). using primers flanking rhe nwlliplc clorung site. and were found to contain inserts that vary in size from 0 Skb to 2.3kb.
The transcript encoding a predominant Trypanosoma evansi variable surface glycoprotein RoTat 1.2 ... more The transcript encoding a predominant Trypanosoma evansi variable surface glycoprotein RoTat 1.2 was cloned and expressed as a recombinant protein in Spodoptera frugiperda and Trichoplusia ni insect cells. Its potential as an antigen for specific detection of antibody in serum of dromedary camels affected by surra, was evaluated. In ELISA, the reactivity of the recombinant RoTat 1.2 VSG was similar to that of native RoTat 1.2 VSG. An indirect agglutination reagent was therefore prepared by coupling the recombinant RoTat 1.2 VSG onto latex particles. The performance of the latex agglutination test was evaluated on camel sera, and compared to the performance of CATT/T. evansi and LATEX/T. evansi tests, using the immune trypanolysis assay with T. evansi RoTat 1.2 as a reference test. The relative sensitivity and specificity of the latex coated with recombinant RoTat 1.2 VSG, using a 1:4 serum dilution, were respectively 89.3 and 99.1%. No differences were observed between the performance of latex coated with recombinant RoTat 1.2 VSG and LATEX/T. evansi or CATT/T. evansi. Here, we describe the successful use of the recombinant RoTat 1.2 VSG for detection of specific antibodies induced by T. evansi infections. pH 7.4). Protein A peroxidase (Sigma) was diluted to 0.2 µg/ml in PBS-Tween and incubated for 1 h (150 µl/well). After 5 washes, wells were incubated for 1 h at ambient temperature with 150 µl ABTS substrate-chromogen solution. The latter was prepared from 50 mg ABTS (2,2'-azinobis(3-ethylbenzothiazoline)-6-sulfonic acid, Boehringer, Germany) dissolved in 100 ml of ABTS-buffer (phosphate-citrate-sodium perborate solution, pH 4.6, Boehringer, Germany). The plate was shaken for 10 s and the OD read at 415 nm (Multiskan RC Version 6.0, Labsystems). Corrected OD values were obtained by subtracting the mean OD of the antigen-free control wells from the mean OD of the corresponding antigen containing wells.
Reduction-substitution reactions on [99mTcO4]- with both o-phenylenebis(dimethylarsine) (DIARS) a... more Reduction-substitution reactions on [99mTcO4]- with both o-phenylenebis(dimethylarsine) (DIARS) and various thiols produce a series of monocationic [99Tc(DIARS)2(SR)2]+ complexes. Addition of [99gTcO4]- to the above reaction mixtures allows the characterization of the "carrier-added" complexes by means of reverse-phase high-performance liquid chromatography with radiometric and optical detection systems. The identity of the [99mTc(DIARS)2(SR)2]+ complexes is confirmed by fast atom bombardment mass spectroscopy; equivalence of the [99gTc(DIARS)2-(SR)2]+ and [99mTc(DIARS)2(SR)2]+ species is demonstrated by identical HPLC retention times. All the [99mTc(DIARS)2(SR)2]+ complexes tested accumulate in the myocardium of Sprague-Dawley rats with an average uptake of 1.5-2.0% of injected dose/g at 30 min. Thus, as designed, these nonreducible Tc(III) complexes do not exhibit the rapid myocardial washout observed for reducible Tc(III) complexes. These [99mTc(DIARS)2(SR)2]+ complexes also exhibit an initially high liver uptake, but the presence of ether groups within the thiolate ligands causes this liver uptake to decrease over time without affecting the heart uptake, thereby improving the heart/liver ratio.
An ~se~richiu coli vector to express and purify foreign proteins by fusion to and separation from... more An ~se~richiu coli vector to express and purify foreign proteins by fusion to and separation from maltose-binding protein (Recombinant DNA; malE; factor X, protease; paramyosin; crosslinked amylose; /$galactosidase; Dirofilaria immi~; affinity purification; over expression; phage rl vectors)
The objective of this work was to evaluate extrusion cooking as a means to improve the nutritiona... more The objective of this work was to evaluate extrusion cooking as a means to improve the nutritional properties of Phaseolus vulgaris L. that had been stored either at 42°C and 80% relative humidity for 6 weeks or for periods >1 year in cereal stores in tropical conditions. Storage under these conditions resulted in an increase in cooking time increased (7.7-and 12-fold, respectively) as a result of development of the hard-to-cook (HTC) defect. Single-screw extrusion of the milled beans was carried out at four barrel temperatures and two moisture contents. The extrudate bulk density and water solubility index decreased with increasing temperature, whereas the water absorption index increased due to the higher proportion of gelatinized starch in the extruded samples. Both fresh and HTC beans contained nutritionally significant amounts of lectins, trypsin, and R-amylase inhibitors, which were mostly inactivated by extrusion. Extrusion also caused a considerable redistribution of insoluble dietary fiber to soluble, although the total dietary fiber content was not affected. Changes in solubility involved pectic polysaccharides, arabinose and uronic acids being the main sugars involved. Stored beans subjected to extrusion cooking showed physical and chemical characteristics similar to those of extrudates from fresh beans. SME (kWh kg -1 ) ) screw speed × power (kW) × torque (%) max screw speed × throughput (kg h -1 ) × 100
The exploitation of ''intelligent'' factory set-ups could enhance the competitiveness of the EU t... more The exploitation of ''intelligent'' factory set-ups could enhance the competitiveness of the EU textile and clothing industries, by enabling collaborative design-and-manufacture options, while achieving economies of scope with effective exploitation of (strategic/tactical/execution) flexibility. Simulation turns to be reference aid for developing and acknowledging the appropriate set-ups and the adaptive-schedules. The investigation, besides of reference concepts, summarises a case example, for explanatory purpose. Combined-mode schedules are considered, showing the benefits for prompt returns and leaving open middle/long horizons issues, still with quantitative checks on alternatives provided by virtual reality tests. #
We examine the contribution of Multiple Parton Interactions to Z + n-jets production at the LHC, ... more We examine the contribution of Multiple Parton Interactions to Z + n-jets production at the LHC, n = 2, 3, 4, where the Z boson is assumed to decay leptonically. We compare the results obtained with the correlated GS09 double parton distribution function with those obtained with two instances of fully factorized single parton distribution functions: MSTW2008LO and CTEQ6L1. It appears quite feasible to measure the MPI contribution to Z + 2/3/4 jets already in the first phase of the LHC with a total luminosity of one inverse femtobarn at 7 TeV. If as expected the trigger threshold for single photons is around 80 GeV, Z + 2-jets production may well turn out to be more easily observable than the γ + 3-jets channel. The MPI cross section is dominated by relatively soft events with two jets balancing in transverse momentum.
There are many possibilities for new physics beyond the Standard Model that feature non-standard ... more There are many possibilities for new physics beyond the Standard Model that feature non-standard Higgs sectors. These may introduce new sources of CP violation, and there may be mixing between multiple Higgs bosons or other new scalar bosons. Alternatively, the Higgs may be a composite state, or there may even be no Higgs at all. These non-standard Higgs scenarios have important implications for collider physics as well as for cosmology, and understanding their phenomenology is essential for a full comprehension of electroweak symmetry breaking. This report discusses the most relevant theories which go beyond the Standard Model and its minimal, CP-conserving supersymmetric extension: two-Higgs-doublet models and minimal supersymmetric models with CP violation, supersymmetric models with an extra singlet, models with extra gauge groups or Higgs triplets, Little Higgs models, models in extra dimensions, and models with technicolour or other new strong dynamics. For each of these scenarios, this report presents an introduction to the phenomenology, followed by contributions on more detailed theoretical aspects and studies of possible experimental signatures at the LHC and other colliders.
Journal of Clinical Endocrinology & Metabolism, 2010
Context: Thyroid hormone, requiring adequate maternal iodine intake, is critical for fetal neurod... more Context: Thyroid hormone, requiring adequate maternal iodine intake, is critical for fetal neurodevelopment. Perchlorate decreases thyroidal iodine uptake by competitively inhibiting the sodium/iodide symporter. It is unclear whether environmental perchlorate exposure adversely affects thyroid function in pregnant women. Thiocyanate, derived from foods and cigarette smoke, is a less potent competitive sodium/iodide symporter inhibitor than perchlorate.
Trypanosoma (Nannomonas) congolense comprises morphologically identical but genetically heterogen... more Trypanosoma (Nannomonas) congolense comprises morphologically identical but genetically heterogeneous parasites infective to livestock and other mammalian hosts; three different genotypes of this parasite have been described previously. Restriction enzyme fragment length polymorphisms (RFLPs) in both kinetoplast DNA minicircle and nuclear DNA sequences, and randomly amplified polymorphic deoxyribonucleic acid (RAPD) patterns have been used here to demonstrate the existence of another type of T. (N.) congolense that is genotypically distinct from those that have so far been characterized at the molecular level. A highly repetitive, tandemly arranged DNA sequence and oligonucleotide primers, for use in polymerase chain reaction (PCR) amplification are described, which can be used for specific identification of the trypanosome and its distinction from others within the Nannomonas subgenus.
The transcript encoding a predominant Trypanosoma evansi variable surface glycoprotein RoTat 1.2 ... more The transcript encoding a predominant Trypanosoma evansi variable surface glycoprotein RoTat 1.2 was cloned and expressed as a recombinant protein in Spodoptera frugiperda and Trichoplusia ni insect cells. Its potential as an antigen for specific detection of antibody in serum of dromedary camels affected by surra, was evaluated. In ELISA, the reactivity of the recombinant RoTat 1.2 VSG was similar to that of native RoTat 1.2 VSG. An indirect agglutination reagent was therefore prepared by coupling the recombinant RoTat 1.2 VSG onto latex particles. The performance of the latex agglutination test was evaluated on camel sera, and compared to the performance of CATT/T. evansi and LATEX/T. evansi tests, using the immune trypanolysis assay with T. evansi RoTat 1.2 as a reference test. The relative sensitivity and specificity of the latex coated with recombinant RoTat 1.2 VSG, using a 1:4 serum dilution, were respectively 89.3 and 99.1%. No differences were observed between the performance of latex coated with recombinant RoTat 1.2 VSG and LATEX/T. evansi or CATT/T. evansi. Here, we describe the successful use of the recombinant RoTat 1.2 VSG for detection of specific antibodies induced by T. evansi infections. pH 7.4). Protein A peroxidase (Sigma) was diluted to 0.2 µg/ml in PBS-Tween and incubated for 1 h (150 µl/well). After 5 washes, wells were incubated for 1 h at ambient temperature with 150 µl ABTS substrate-chromogen solution. The latter was prepared from 50 mg ABTS (2,2'-azinobis(3-ethylbenzothiazoline)-6-sulfonic acid, Boehringer, Germany) dissolved in 100 ml of ABTS-buffer (phosphate-citrate-sodium perborate solution, pH 4.6, Boehringer, Germany). The plate was shaken for 10 s and the OD read at 415 nm (Multiskan RC Version 6.0, Labsystems). Corrected OD values were obtained by subtracting the mean OD of the antigen-free control wells from the mean OD of the corresponding antigen containing wells.
Two clones of T. congolenre were derived. one from the other. by growth in mice and administralmn... more Two clones of T. congolenre were derived. one from the other. by growth in mice and administralmn of increasing sub-curat~ve doses of lhe Qpanocldc isometamidiom chloride. One of the clones was Ihus made approunatel! IO more tolerant to lhe bypanocide than the other mP.NA from lhc Iwo clones was used for syntbeas of co~npleu~enta~ DNA (cDNA) An aliquol of the cDNA was convened inlo double-stranded DNA molecules and Qrccl~onall) ligated 8 the EwRIiNotl she of hgt I I. which were subscqucntly espandcd by growih m Ecoli Y IOLX). The libraries obtamed had >I()' independenl phage clones, >90% of which were recombinant. One hundred random recomtxnant clones from each library were subjected to the polymcrase chao reaction (F'CR). using primers flanking rhe nwlliplc clorung site. and were found to contain inserts that vary in size from 0 Skb to 2.3kb.
The transcript encoding a predominant Trypanosoma evansi variable surface glycoprotein RoTat 1.2 ... more The transcript encoding a predominant Trypanosoma evansi variable surface glycoprotein RoTat 1.2 was cloned and expressed as a recombinant protein in Spodoptera frugiperda and Trichoplusia ni insect cells. Its potential as an antigen for specific detection of antibody in serum of dromedary camels affected by surra, was evaluated. In ELISA, the reactivity of the recombinant RoTat 1.2 VSG was similar to that of native RoTat 1.2 VSG. An indirect agglutination reagent was therefore prepared by coupling the recombinant RoTat 1.2 VSG onto latex particles. The performance of the latex agglutination test was evaluated on camel sera, and compared to the performance of CATT/T. evansi and LATEX/T. evansi tests, using the immune trypanolysis assay with T. evansi RoTat 1.2 as a reference test. The relative sensitivity and specificity of the latex coated with recombinant RoTat 1.2 VSG, using a 1:4 serum dilution, were respectively 89.3 and 99.1%. No differences were observed between the performance of latex coated with recombinant RoTat 1.2 VSG and LATEX/T. evansi or CATT/T. evansi. Here, we describe the successful use of the recombinant RoTat 1.2 VSG for detection of specific antibodies induced by T. evansi infections. pH 7.4). Protein A peroxidase (Sigma) was diluted to 0.2 µg/ml in PBS-Tween and incubated for 1 h (150 µl/well). After 5 washes, wells were incubated for 1 h at ambient temperature with 150 µl ABTS substrate-chromogen solution. The latter was prepared from 50 mg ABTS (2,2'-azinobis(3-ethylbenzothiazoline)-6-sulfonic acid, Boehringer, Germany) dissolved in 100 ml of ABTS-buffer (phosphate-citrate-sodium perborate solution, pH 4.6, Boehringer, Germany). The plate was shaken for 10 s and the OD read at 415 nm (Multiskan RC Version 6.0, Labsystems). Corrected OD values were obtained by subtracting the mean OD of the antigen-free control wells from the mean OD of the corresponding antigen containing wells.
Trypanosoma (Nannomonas) congolense comprises morphologically identical but genetically heterogen... more Trypanosoma (Nannomonas) congolense comprises morphologically identical but genetically heterogeneous parasites infective to livestock and other mammalian hosts; three different genotypes of this parasite have been described previously. Restriction enzyme fragment length polymorphisms (RFLPs) in both kinetoplast DNA minicircle and nuclear DNA sequences, and randomly amplified polymorphic deoxyribonucleic acid (RAPD) patterns have been used here to demonstrate the existence of another type of T. (N.) congolense that is genotypically distinct from those that have so far been characterized at the molecular level. A highly repetitive, tandemly arranged DNA sequence and oligonucleotide primers, for use in polymerase chain reaction (PCR) amplification are described, which can be used for specific identification of the trypanosome and its distinction from others within the Nannomonas subgenus.
The transcript encoding a predominant Trypanosoma evansi variable surface glycoprotein RoTat 1.2 ... more The transcript encoding a predominant Trypanosoma evansi variable surface glycoprotein RoTat 1.2 was cloned and expressed as a recombinant protein in Spodoptera frugiperda and Trichoplusia ni insect cells. Its potential as an antigen for specific detection of antibody in serum of dromedary camels affected by surra, was evaluated. In ELISA, the reactivity of the recombinant RoTat 1.2 VSG was similar to that of native RoTat 1.2 VSG. An indirect agglutination reagent was therefore prepared by coupling the recombinant RoTat 1.2 VSG onto latex particles. The performance of the latex agglutination test was evaluated on camel sera, and compared to the performance of CATT/T. evansi and LATEX/T. evansi tests, using the immune trypanolysis assay with T. evansi RoTat 1.2 as a reference test. The relative sensitivity and specificity of the latex coated with recombinant RoTat 1.2 VSG, using a 1:4 serum dilution, were respectively 89.3 and 99.1%. No differences were observed between the performance of latex coated with recombinant RoTat 1.2 VSG and LATEX/T. evansi or CATT/T. evansi. Here, we describe the successful use of the recombinant RoTat 1.2 VSG for detection of specific antibodies induced by T. evansi infections. pH 7.4). Protein A peroxidase (Sigma) was diluted to 0.2 µg/ml in PBS-Tween and incubated for 1 h (150 µl/well). After 5 washes, wells were incubated for 1 h at ambient temperature with 150 µl ABTS substrate-chromogen solution. The latter was prepared from 50 mg ABTS (2,2'-azinobis(3-ethylbenzothiazoline)-6-sulfonic acid, Boehringer, Germany) dissolved in 100 ml of ABTS-buffer (phosphate-citrate-sodium perborate solution, pH 4.6, Boehringer, Germany). The plate was shaken for 10 s and the OD read at 415 nm (Multiskan RC Version 6.0, Labsystems). Corrected OD values were obtained by subtracting the mean OD of the antigen-free control wells from the mean OD of the corresponding antigen containing wells.
Two clones of T. congolenre were derived. one from the other. by growth in mice and administralmn... more Two clones of T. congolenre were derived. one from the other. by growth in mice and administralmn of increasing sub-curat~ve doses of lhe Qpanocldc isometamidiom chloride. One of the clones was Ihus made approunatel! IO more tolerant to lhe bypanocide than the other mP.NA from lhc Iwo clones was used for syntbeas of co~npleu~enta~ DNA (cDNA) An aliquol of the cDNA was convened inlo double-stranded DNA molecules and Qrccl~onall) ligated 8 the EwRIiNotl she of hgt I I. which were subscqucntly espandcd by growih m Ecoli Y IOLX). The libraries obtamed had >I()' independenl phage clones, >90% of which were recombinant. One hundred random recomtxnant clones from each library were subjected to the polymcrase chao reaction (F'CR). using primers flanking rhe nwlliplc clorung site. and were found to contain inserts that vary in size from 0 Skb to 2.3kb.
The transcript encoding a predominant Trypanosoma evansi variable surface glycoprotein RoTat 1.2 ... more The transcript encoding a predominant Trypanosoma evansi variable surface glycoprotein RoTat 1.2 was cloned and expressed as a recombinant protein in Spodoptera frugiperda and Trichoplusia ni insect cells. Its potential as an antigen for specific detection of antibody in serum of dromedary camels affected by surra, was evaluated. In ELISA, the reactivity of the recombinant RoTat 1.2 VSG was similar to that of native RoTat 1.2 VSG. An indirect agglutination reagent was therefore prepared by coupling the recombinant RoTat 1.2 VSG onto latex particles. The performance of the latex agglutination test was evaluated on camel sera, and compared to the performance of CATT/T. evansi and LATEX/T. evansi tests, using the immune trypanolysis assay with T. evansi RoTat 1.2 as a reference test. The relative sensitivity and specificity of the latex coated with recombinant RoTat 1.2 VSG, using a 1:4 serum dilution, were respectively 89.3 and 99.1%. No differences were observed between the performance of latex coated with recombinant RoTat 1.2 VSG and LATEX/T. evansi or CATT/T. evansi. Here, we describe the successful use of the recombinant RoTat 1.2 VSG for detection of specific antibodies induced by T. evansi infections. pH 7.4). Protein A peroxidase (Sigma) was diluted to 0.2 µg/ml in PBS-Tween and incubated for 1 h (150 µl/well). After 5 washes, wells were incubated for 1 h at ambient temperature with 150 µl ABTS substrate-chromogen solution. The latter was prepared from 50 mg ABTS (2,2'-azinobis(3-ethylbenzothiazoline)-6-sulfonic acid, Boehringer, Germany) dissolved in 100 ml of ABTS-buffer (phosphate-citrate-sodium perborate solution, pH 4.6, Boehringer, Germany). The plate was shaken for 10 s and the OD read at 415 nm (Multiskan RC Version 6.0, Labsystems). Corrected OD values were obtained by subtracting the mean OD of the antigen-free control wells from the mean OD of the corresponding antigen containing wells.
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