Detection of cross infections by Leishmania spp. and Trypanosoma spp. in dogs using indirect immunoenzyme assay, indirect fluorescent antibody test and polymerase chain reaction

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Parentage testing in alpacas (Vicugna pacos) using semi-automated fluorescent multiplex PCRs with 10 microsatellite markers

The aim of this study was to assess and apply a microsatellite multiplex system for parentage determination in alpacas. An approach for parentage testing based on 10 microsatellites was evaluated in a population of 329 unrelated alpacas from different geographical zones in Peru. All microsatellite markers, which amplified in two multiplex reactions, were highly polymorphic with a mean of 14.5 alleles per locus (six to 28 alleles per locus) and an average expected heterozygosity (H-E) of 0.8185 (range of 0.698-0.946). The total parentage exclusion probability was 0.999456 for excluding a candidate parent from parentage of an arbitrary offspring, given only the genotype of the offspring, and 0.999991 for excluding a candidate parent from parentage of an arbitrary offspring, given the genotype of the offspring and the other parent. In a case test of parentage assignment, the microsatellite panel assigned 38 (from 45 cases) offspring parentage to 10 sires with LOD scores ranging from 2.19 x 10(+13) to 1.34 x 10(+15) and Delta values ranging from 2.80 x 10(+12) to 1.34 x 10(+15) with an estimated pedigree error rate of 15.5%. The performance of this multiplex panel of markers suggests that it will be useful in parentage testing of alpacas.

Use of In Vivo and In Vitro Systems to Select Leishmania amazonensis Expressing Green Fluorescent Protein

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Indirect fluorescent test for detection of anti-Paracoccidioides brasiliensis antibodies using coated bentonite particles as antigen

An indirect fluorescent test was developed for detecting antibodies to Paracoccidioides brasiliensis using bentonite particles as antigen (Bent-IF). The bentonite particles were coated with P. brasiliensis polysaccharide antigen and tested with sera from paracoccidioidomycosis patients (36 sera), normal blood donors (32 sera) and patients with non-mycotic diseases (29 sera). The titres given by the positive sera were compared with those of complement fixation (CF), immunodiffusion (ID) and immunofluorescent test using yeast forms of the fungus as antigen (conventional-IF). All normal blood donors' sera gave a negative Bent-IF, conventional-IF, ID and CF tests. All paracoccidioidomycosis sera were reactive in conventional-IF and gave concordant results in Bent-IF. There was no correlation between CF and Bent-IF titres. 27·6% of sera from patients with non-mycotic diseases gave weak titres in both IF-tests. The present data indicate that the Bent-IF is a sensitive and simple serodiagnostic technique comparable with the conventional P. brasiliensis antibody test. © 1983.

Novel high-power-factor isolated electronic ballast for multiple tubular fluorescent lamps

This paper presents a novel isolated electronic ballast for multiple fluorescent lamps, featuring high power-factor, and high efficiency. Two stages compose this new electronic ballast, namely, a new voltage step-down isolated Sepic rectifier, and a classical resonant Half-Bridge inverter. The new isolated Sepic rectifier is obtained from a Zero-Current-Switching (ZCS) Pulse-Width-Modulated (PWM) soft-commutation cell. The average-current control technique is used in this preregulator stage in order to provide low phase displacement and low Total-Harmonic-Distortion (THD) at input current, resulting in high power-factor, and attending properly IEC 61000-3-2 standards. The resonant Half-Bridge inverter performs Zero-Voltage-Switching (ZVS), providing conditions for the obtaining of overall high efficiency. It is developed a design example for the new isolated electronic ballast rated at 200W output power, 220Vrms input voltage, 115Vdc dc link voltage, with rectifier and inverter stages operating at 50kHz. Finally, experimental results are presented in order to verify the developed analysis. The THD at input current is equal to 5.25%, for an input voltage THD equal to 1.63%, and the measured overall efficiency is about 88.25%, at rated load.

Two short peptides including segments of subunit A of Escherichia coli DNA gyrase as potential probes to evaluate the antibacterial activity of quinolones

Quinolones constitute a family of compounds with a potent antibiotic activity. The enzyme DNA gyrase, responsible for the replication and transcription processes in DNA of bacteria, is involved in the mechanism of action of these drugs. In this sense, it is believed that quinolones stabilize the so-called 'cleavable complex' formed by DNA and gyrase, but the whole process is still far from being understood at the molecular level. This information is crucial in order to design new biological active products. As an approach to the problem, we have designed and synthesized low molecular weight peptide mimics of DNA gyrase. These peptides correspond to sequences of the subunit A of the enzyme from Escherichia coli, that include the quinolone resistance-determining region (positions 75-92) and a segment containing the catalytic Tyr-122 (positions 116-130). The peptide mimic of the non-mutated enzyme binds to ciprofloxin (CFX) only when DNA and Mg2+ were present (Kd = 1.6 × 10 -6 m), a result previously found with DNA gyrase. On the other hand, binding was reduced when mutations of Ser-83 to Leu-83 and Asp-87 to Asn-87 were introduced, a double change previously found in the subunit A of DNA gyrase from several CFX-resistant clinical isolates of E. coli. These results suggest that synthetic peptides designed in a similar way to that described here can be used as mimics of gyrases (topoisomerases) in order to study the binding of the quinolone to the enzyme-DNA complex as well as the mechanism of action of these antibiotics. Copyright © 2001 European Peptide Society and John Wiley & Sons, Ltd.

A high efficiency HPF-ZCS-PWM Sepic for electronic ballast with multiple tubular fluorescent lamps

This paper presents a high efficiency Sepic rectifier for an electronic ballast application with multiple fluorescent lamps. The proposed Sepic rectifier is based on a Zero-Current-Switching (ZCS) Pulse-Width-Modulated (PWM) soft-commutation cell. The high power-factor of this structure is obtained using the instantaneous average-current control technique, in order to attend properly IEC61000-3-2 standards. The inverting stage of this new electronic ballast is a classical Zero-Voltage-Switching (ZVS) Half-Bridge inverter. A proper design methodology is developed for this new electronic ballast, and a design example is presented for an application with five fluorescent lamps 40W-T12 (200W output power), 220Vrms input voltage, 130Vdc dc link voltage, with rectifier and inverter stages operating at 50kHz. Experimental results are also presented. The THD at input current is equal to 6.41%, for an input voltage THD equal to 2.14%, and the measured overall efficiency is about 92.8%, at rated load.

Short interspersed repetitive elements (SINEs) from the cichlid fish, Oreochromis niloticus, and their chromosomal localization by fluorescent in situ hybridization

In the present study, we describe the cloning and characterization of a new SINE-like element from O. niloticus (ROn-2) and show the distribution of this SINE and a previously isolated SINE, ROn-1, in the chromosomes of O. niloticus. The ROn-2 element is 359 base pairs (bp) in length, contains short direct terminal repeats, a tRNA-related region similar to tRNA Val and tRNA Arg, a tRNA-unrelated region, and a poly-A tail. Analysis of the chromosomal distribution of ROn-1 and ROn-2 by fluorescent in situ hybridization showed that both SINE sequences are present in all chromosomes of tilapia, and organized in small clusters. The only exception was a large cluster of ROn-1 repeats found in the middle of the long arm of chromosome 1. In view of our data we discuss the hypothesis that the absence of large clusters of SINE sequences and the structural composition of these sequences may explain the absence of base-specific fluorochrome bands in the chromosomes of tilapia.

An Improved Design Procedure for LCC Resonant Filter of Dimmable Electronic Ballasts for Fluorescent Lamps, Based on Lamp Model

This paper presents an improved design methodology for determining the parameters used in the classical Series-Parallel Loaded Resonant (SPLR) filter employed in the switching frequency controlled dimmable electronic ballasts. According to the analysis developed in this paper, it is possible to evaluate some characteristics of the resonant filter during the dimming process, such as: range of switching frequency, phase shift and rms value of the current drained by the resonant filter + fluorescent lamp set.

Dimmable electronic ballast with high power-factor SEPIC preregulator, for multiple tubular fluorescent lamps

An electronic ballast for multiple tubular fluorescent lamps is presented in this paper. The proposed structure features high power-factor, dimming capability, and soft-switching to the semiconductor devices operated in high frequencies. A Zero-Current-Switching - Pulse-Width-Modulated (ZCS-PWM) SEPIC converter composes the rectifying stage, controlled by the instantaneous average input current technique, performing soft-commutations and high input power factor. Regarding the inverting stage, it is composed by a classical resonant Half-Bridge converter, associated to Series Parallel-Loaded Resonant (SPLR) filters. The dimming control technique employed in this Half-Bridge inverter is based on the phase-shift in the current processed through the sets of filter + lamp. In addition, experimental results are shown in order to validate the developed analysis.

An improved design procedure for LCC resonant filter of dimmable electronic ballasts for fluorescent lamps, based on lamp model

This paper presents an improved design methodology for the determination of the parameters used in the classical series-resonant parallel-loaded (SRPL) filter employed in the switching frequency controlled dimmable electronic ballasts. According to the analysis developed in this paper, it is possible to evaluate some important characteristics of the resonant filter during the dimming operation, such as: range of switching frequency, phase shift, and rms value of the current drained by the resonant filter + fluorescent lamp set. Experimental results are presented in order to validate the analyses developed in this paper. © 2005 IEEE.