Development of a Loop-Mediated Isothermal Amplification Method for Detection of Histoplasma capsulatum DNA in Clinical Samples

Improved methods for the detection of Histoplasma capsulatum are needed in regions with limited resources in which the organism is endemic, where delayed diagnosis of progressive disseminated histoplasmosis (PDH) results in high mortality rates. We have investigated the use of a loop-mediated isothermal amplification (LAMP) assay to facilitate rapid inexpensive molecular diagnosis of this disease. Primers for LAMP were designed to amplify the Hcp100 locus of H. capsulatum. The sensitivity and limit of detection were evaluated using DNA extracted from 91 clinical isolates of known geographic subspecies, while the assay specificity was determined using DNA extracted from 50 other fungi and Mycobacterium tuberculosis. Urine specimens (n = 6) collected from HIV-positive individuals with culture- and antigen-proven histoplasmosis were evaluated using the LAMP assay. Specimens from healthy persons (n = 10) without evidence of histoplasmosis were used as assay controls. The Hcp100 LAMP assay was 100% sensitive and specific when tested with DNA extracted from culture isolates. The median limit of detection was <= 6 genomes (range, 1 to 300 genomes) for all except one geographic subspecies. The LAMP assay detected Hcp100 in 67% of antigen-positive urine specimens (4/6 specimens), and results were negative for Hcp100 in all healthy control urine specimens. We have shown that the Hcp100 LAMP assay is a rapid affordable assay that can be used to expedite culture confirmation of H. capsulatum in regions in which PDH is endemic. Further, our results indicate proof of the concept that the assay can be used to detect Histoplasma DNA in urine. Further evaluation of this assay using body fluid samples from a larger patient population is warranted.

Time-periodic perturbation of a Lienard equation with an unbounded homoclinic loop

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Effect of oxidizing atmosphere on the electrical properties of SrBi4Ti4O15 thin films obtained by the polymeric precursor method

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

Variability of the conserved V3 loop tip motif in HIV-1 subtype B isolates collected from Brazilian and French patients

The diversity of the V3 loop tip motif sequences of HIV-1 subtype B was analyzed in patients from Botucatu (Brazil) and Montpellier (France). Overall, 37 tetrameric tip motifs were identified, 28 and 17 of them being recognized in Brazilian and French patients, respectively. The GPGR (P) motif was predominant in French but not in Brazilian patients (53.5% vs 31.0%), whereas the GWGR (W) motif was frequent in Brazilian patients (23.0%) and absent in French patients. Three tip motif groups were considered: P, W, and non-P non-W groups. The distribution of HIV-1 isolates into the three groups was significantly different between isolates from Botucatu and from Montpellier (P < 0.001). A higher proportion of CXCR4-using HIV-1 (X4 variants) was observed in the non-P non-W group as compared with the P group (37.5% vs 19.1%), and no X4 variant was identified in the W group (P < 0.001). The higher proportion of X4 variants in the non-P non-W group was essentially observed among the patients from Montpellier, who have been infected with HIV-1 for a longer period of time than those from Botucatu. Among patients from Montpellier, CD4+ cell counts were lower in patients belonging to the non-P non-W group than in those belonging to the P group (24 cells/µL vs 197 cells/µL; P = 0.005). Taken together, the results suggest that variability of the V3 loop tip motif may be related to HIV-1 coreceptor usage and to disease progression. However, as analyzed by a bioinformatic method, the substitution of the V3 loop tip motif of the subtype B consensus sequence with the different tip motifs identified in the present study was not sufficient to induce a change in HIV-1 coreceptor usage.

Massless and massive one-loop three-point functions in negative dimensional approach

In this article we present the complete massless and massive one-loop triangle diagram results using the negative dimensional integration method (NDIM). We consider the following cases: massless internal fields; one massive, two massive with the same mass m and three equal masses for the virtual particles. Our results are given in terms of hypergeometric and hypergeometric-type functions of the external momenta (and masses for the massive cases) where the propagators in the Feynman integrals are raised to arbitrary exponents and the dimension of the space-time is D. Our approach reproduces the known results; it produces other solutions as yet unknown in the literature as well. These new solutions occur naturally in the context of NDIM revealing a promising technique to solve Feynman integrals in quantum field theories.

NDIM achievements: massive, arbitrary tensor rank and N-loop insertions in Feynman integrals

One of the main difficulties in studying quantum field theory, in the perturbative regime, is the calculation of D-dimensional Feynman integrals. In general, one introduces the so-called Feynman parameters and, associated with them, the cumbersome parametric integrals. Solving these integrals beyond the one-loop level can be a difficult task. The negative-dimensional integration method (NDIM) is a technique whereby such a problem is dramatically reduced. We present the calculation of two-loop integrals in three different cases: scalar ones with three different masses, massless with arbitrary tensor rank, with and N insertions of a two-loop diagram.

Phase-Locked Loop design applied to frequency-modulated atomic force microscope

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

Potassium niobate thin films prepared through polymeric precursor method

Thin films of potassium niobate were deposited on (100) Si substrates by the polymeric precursor method (Pechini method). Annealing in static air was performed at 600degrees C for 20 h. The obtained films were characterized by X-ray diffraction and atomic force microscopy (AFM). Electrical characterization of the films pointed to ferroelectricity via hysteresis loop. The dielectric constant, dissipation factor and resistance were measured in frequency region from 10 Hz to 10 MHz. At 1 MHz, the dielectric constant was 158 and the dissipation factor was 0.11. (C) 2004 Elsevier B.V. All rights reserved.

LiNbO3 thin films prepared through polymeric precursor method

Thin films of lithium niobate were deposited on (100) silicon by the polymeric precursor method (Pechini method). Annealing in static air was performed at 500degreesC for 3 h. The films obtained were characterized by X-ray diffraction, scanning electron microscopy and transmission electron microscopy. Electrical characterization of the films pointed to ferroelectricity via hysteresis loop. The dielectric constant, dissipation factor and resistance were measured in the frequency region from 10 Hz to 10 MHz. At 1 MHz, the dielectric constant was 46 and the dissipation factor was 0.043. (C) 2002 Elsevier B.V. B.V. All rights reserved.

Microstructural and ferroelectric properties of PbZr1-xTi(x)O(3) thin films prepared by the polymeric precursor method

The polymeric precursor method was employed in the preparation of PZT thin films on Pt(111)Ti/SiO2/Si(100) substrates. X-ray diffraction patterns revealed the polycrystalline nature of the PZT (53:47) thin films, which had a granular structure and a grain size of approximately 70 nm. A 350-nm thick film was obtained by running three cycles of the dip-coating/heating process. Atomic force microscopy (AFM) analyses showed the surface of these thin films to be smooth, dense and crack-free with low surface roughness (= 2.0 nm). The PZT (53:47) thin films annealed at 700 degreesC showed a well-saturated hysteresis loop. The C-V curves of perovskite thin film displayed normal ferroelectric behavior, while the remanent polarization (2P(r)) and coercive field (E-e) of the film deposited and measured at room temperature were 40 muC/cm(2) and 110 kV/cm, respectively. (C) 2001 Elsevier B.V. B.V. All rights reserved.

Oscillating holograms recorded in photorefractive crystals by a frequency detuned feedback loop

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

Two-loop self-energy diagrams worked out with NDIM

In this work we calculate two two-loop massless Feynman integrals pertaining to self-energy diagrams using NDIM (Negative Dimensional Integration Method). We show that the answer we get is 36-fold degenerate. We then consider special cases of exponents for propagators and the outcoming results compared with known ones obtained via traditional methods.

Probing negative-dimensional integration: Two-loop covariant vertex and one-loop light-cone integrals

The negative-dimensional integration method (NDIM) seems to be a very promising technique for evaluating massless and/or massive Feynman diagrams. It is unique in the sense that the method gives solutions in different regions of external momenta simultaneously. Moreover, it is a technique whereby the difficulties associated with performing parametric integrals in the standard approach are transferred to a simpler solving of a system of linear algebraic equations, thanks to the polynomial character of the relevant integrands. We employ this method to evaluate a scalar integral for a massless two-loop three-point vertex with all the external legs off-shell, and consider several special cases for it, yielding results, even for distinct simpler diagrams. We also consider the possibility of NDIM in non-covariant gauges such as the light-cone gauge and do some illustrative calculations, showing that for one-degree violation of covariance (i.e. one external, gauge-breaking, light-like vector n μ) the ensuing results are concordant with the ones obtained via either the usual dimensional regularization technique, or the use of the principal value prescription for the gauge-dependent pole, while for two-degree violation of covariance - i.e. two external, light-like vectors n μ, the gauge-breaking one, and (its dual) n * μ - the ensuing results are concordant with the ones obtained via causal constraints or the use of the so-called generalized Mandelstam-Leibbrandt prescription. © 1999 Elsevier Science B.V.

Loop Integrals in Three Outstanding Gauges: Feynman, Light-Cone, and Coulomb

We apply the negative dimensional integration method (NDIM) to three outstanding gauges: Feynman, light-cone, and Coulomb gauges. Our aim is to show that NDIM is a very suitable technique to deal with loop integrals, regardless of which gauge choice that originated them. In the Feynman gauge we perform scalar two-loop four-point massless integrals; in the light-cone gauge we calculate scalar two-loop integrals contributing to two-point functions without any kind of prescriptions, since NDIM can abandon such devices - this calculation is the first test of our prescriptionless method beyond one-loop order; and finally, for the Coulomb gauge we consider a four-propagator massless loop integral, in the split-dimensional regularization context. © 2001 Academic Press.

On the development of the electrochemical potentiokinetic method

The development of electrochemical potentiokinetic methods as applied to the testing of metals and alloys is followed from its early phases up to its latest advances relating to intergranular corrosion, SCC and pitting corrosion tests of stainless steels and special alloys and to the examination of their structure and properties. In assessing the susceptibility to intergranular and pitting corrosion by potentiokinetic polarization tests, the polarization curves which apply to the bulk of the alloy grains (the matrix) must be distinguished from those pertaining to grain boundaries. Cyclic polarization measurements such as the electrochemical potentiokinetic reactivation (EPR) test make it possible to derive the alloy's susceptibility to intergranular, pitting and crevice corrosion from characteristic potentials and other quantities determined in the 'double loop' test. EPR is rapid and responds to the combined effects of a number of factors that influence the properties of materials. The electrochemical p otentiokinetic tests are sensitive enough to detect structural changes in heat treated materials ranging far beyond the stainless steels alone, and can be used for non-destructive testing aimed at elucidating the properties and behavior of materials. © 2001 Elsevier Science Ltd. All rights reserved.