336 research outputs found
Simultaneous detection of the C282Y, H63D and S65C mutations in the hemochromatosis gene using quenched-FRET real-time PCR
Hereditary hemochromatosis (HH) is a common autosomal disorder of iron metabolism mainly affecting Caucasian populations. Three recurrent disease-associated mutations have been detected in the hemochromatosis gene (HFE): C282Y, H63D, and S65C. Although HH phenotype has been associated with all three mutations, C282Y is considered the most relevant mutation responsible for hemochromatosis. Clinical complications of HH include cirrhosis of the liver, congestive cardiac failure and cardiac arrhythmias, endocrine pancreatic disease, which can be prevented by early diagnosis and treatment. Therefore, a reliable genotyping method is required for presymptomatic diagnosis. We describe the simultaneous detection of the C282Y, H63D and S65C mutations in the hemochromatosis gene by real-time PCR followed by melting curve analysis using fluorescence resonance energy transfer (FRET) probes. The acceptor fluorophore may be replaced by a quencher, increasing multiplex possibilities. Real-time PCR results were compared to the results of sequencing and conventional PCR followed by restriction digestion and detection by agarose gel electrophoresis (PCR-RFLP). Genotypes from 80 individuals obtained both by the conventional PCR-RFLP method and quenched-FRET real-time PCR were in full agreement. Sequencing also confirmed the results obtained by the new method, which proved to be an accurate, rapid and cost-effective diagnostic assay. Our findings demonstrate the usefulness of real-time PCR for the simultaneous detection of mutations in the HFE gene, which allows a reduction of a significant amount of time in sample processing compared to the PCR-RFLP method, eliminates the use of toxic reagents, reduces the risk of contamination in the laboratory, and enables full process automation
Massive quantum fields in a conical background
Representations of the Klein-Gordon and Dirac propagators are determined in a
dimensional conical background for massive fields twisted by an arbitrary
angle . The Dirac propagator is shown to be obtained from the
Klein-Gordon propagator twisted by angles where
is the cone deficit angle. Vacuum expectation values are determined
by a point-splitting method in the proper time representation of the
propagators. Analogies with the Aharonov-Bohm effect are pointed out throughout
the paper and a conjecture on an extension to fields of arbitrary spin is
given.Comment: Propagator (11) is rewritten in a more convenient form and the one
before that is amended. A more concise expression is given for the energy
density of a twisted spinor. The references contain minor correction
Charge Fluctuations and Counterion Condensation
We predict a condensation phenomenon in an overall neutral system, consisting
of a single charged plate and its oppositely charged counterions. Based on the
``two-fluid'' model, in which the counterions are divided into a ``free'' and a
``condensed'' fraction, we argue that for high surface charge, fluctuations can
lead to a phase transition in which a large fraction of counterions is
condensed. Furthermore, we show that depending on the valence, the condensation
is either a first-order or a smooth transition.Comment: 16 pages, 1 figure, accepted to be published in PR
Counterion Condensation and Fluctuation-Induced Attraction
We consider an overall neutral system consisting of two similarly charged
plates and their oppositely charged counterions and analyze the electrostatic
interaction between the two surfaces beyond the mean-field Poisson-Boltzmann
approximation. Our physical picture is based on the fluctuation-driven
counterion condensation model, in which a fraction of the counterions is
allowed to ``condense'' onto the charged plates. In addition, an expression for
the pressure is derived, which includes fluctuation contributions of the whole
system. We find that for sufficiently high surface charges, the distance at
which the attraction, arising from charge fluctuations, starts to dominate can
be large compared to the Gouy-Chapmann length. We also demonstrate that
depending on the valency, the system may exhibit a novel first-order binding
transition at short distances.Comment: 15 pages, 8 figures, to appear in PR
Euclidean Approach to the Entropy for a Scalar Field in Rindler-like Space-Times
The off-shell entropy for a massless scalar field in a D-dimensional
Rindler-like space-time is investigated within the conical Euclidean approach
in the manifold C_\be\times\M^N, C_\be being the 2-dimensional cone, making
use of the zeta-function regularisation. Due to the presence of conical
singularities, it is shown that the relation between the zeta-function and the
heat kernel is non trivial and, as first pointed out by Cheeger, requires a
separation between small and large eigenvalues of the Laplace operator. As a
consequence, in the massless case, the (naive) non existence of the Mellin
transform is by-passed by the Cheeger's analytical continuation of the
zeta-function on manifold with conical singularities. Furthermore, the
continuous spectrum leads to the introduction of smeared traces. In general, it
is pointed out that the presence of the divergences may depend on the smearing
function and they arise in removing the smearing cutoff. With a simple choice
of the smearing function, horizon divergences in the thermodynamical quantities
are recovered and these are similar to the divergences found by means of
off-shell methods like the brick wall model, the optical conformal
transformation techniques or the canonical path integral method.Comment: 17 pages, LaTex. A sign error corrected and few comments adde
Aspectos epidemiológicos da leishmaniose visceral canina no município de Pedro Leopoldo, Minas Gerais, 2003
Efeito da Gliricidia sepium sobre nutrientes do solo, microclima e produtividade do milho em sistema agroflorestal no Agreste Paraibano.
Gliricidia sepium é uma leguminosa arbórea que tem sido utilizada em sistemas em aléias no semi-árido nordestino por apresentar bom desenvolvimento em condições de estresse hídrico. Entretanto, há pouca informação disponível sobre o efeito da introdução dessa espécie nos agroecossistemas da região. No presente estudo, objetivou-se avaliar a influência da distância de plantas de Gliricidia sepium sobre características da cultura do milho e do solo e microclima no Agreste Paraibano. O estudo foi realizado no município de Esperança (PB), em área de 0,5 ha, onde, em 1996, foram plantadas fileiras de G. sepium espaçadas 6 m entre si e com 1 m entre as árvores. Nesta área, em 2002, foram delimitadas quatro parcelas de 6 x 8 m e, em cada parcela, foi estabelecido um transeto perpendicular às fileiras de árvores com três posições de amostragem: (1) nas fileiras de árvores (0 m); (2) a 1 m das fileiras de árvores, e (3) a 3 m de distância das fileiras de árvores. O delineamento experimental utilizado foi em blocos casualizados com quatro repetições. A massa seca de folhedo caído embaixo da fileira de árvores foi de 1.390 kg ha-1 e diminuiu, gradativamente, para 270 kg ha-1 a 3 m de distância das árvores. As concentrações de P, K e matéria orgânica leve (MOL) embaixo das árvores foram maiores do que a 1 e 3 m de distância das fileiras. As médias mensais das temperaturas mínimas do ar e do solo embaixo e a 3 m das árvores foram similares. Entretanto, as médias mensais das temperaturas máximas do solo e do ar foram de 6 e 2 °C mais altas a 3 m das árvores, respectivamente, ao longo do período de estudo. A umidade do solo foi significativamente menor embaixo das árvores do que a 1 e 3 m de distância. O milho produziu mais grãos e palha e acumulou mais nutrientes nas posições mais próximas das fileiras de G. sepium
Cover plants and mineral nitrogen: effects on organic matter fractions in an oxisol under no-tillage in the cerrado
Efeito da idade da matriz sobre o desempenho e rendimento de carcaça de frangos de corte
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