Inflammatory cytokine mRNA detection by real time PCR in chorioamniotic membranes from pregnant women with preterm premature rupture of membranes

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

Development of a real-time PCR method for rapid sexing of human preimplantation embryos

Genes on the X chromosome are known to be responsible for more than 200 hereditary diseases. After IVF, the simple selection of embryo sex before uterine transfer can prevent the occurrence of affected offspring among couples at risk for these genetic disorders. The aim of this investigation was to develop a rapid method of preimplantation genetic diagnosis (PGD) using real-time polymerase chain reaction (PCR) for the sexing of human embryos, and to compare it to the fluorescence in-situ hybridization technique, considered to be the gold standard. After biopsies were obtained from 40 surplus non-viable embryos for transfer, a total of 98 blastomeres were analysed. It was possible to analyse 24 embryos (60%) by both techniques, generating a total of 70 blastomeres (35 per technique), white 28 blastomeres from 16 embryos (40%) were analysed only by real-time PCR. A rapid and safe method was developed in the present study for the sexual diagnosis of a single human cell (blastomere and buccal cell) using the emerging technology of real-time PCR. (C) 2009, Reproductive Healthcare Ltd. Published by Elsevier Ltd. All rights reserved.

Cloning, sequencing and expression analysis of the equine hepcidin gene by real-time PCR

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

Real-time ultrasound to predict rabbit carcass composition and volume of longissimus dorsi muscle

Real-time ultrasonography (RTU) was used to measure the longissimus dorsi muscle (LM) volume in vivo and to predict the carcass composition of rabbits. For this, 63 New Zealand White × Californian rabbits with 2093±63 g live weight were used. Animals were scanned between the 6th and 7th lumbar vertebrae using an RTU equipment with a 7.5 MHz probe. Measurements of LM volume were obtianed both in vivo and on carcass. Regression equations were used for the prediction of carcass composition and LM volume using the LM volume measured obtained with RTU (LMVU) as independent variable. Carcass meat, bone and total dissectible fat weights represented 780, 164 and 56 g/kg of the reference carcass weight, respectively. Regression equations showed a strong relationship between LMVU and the correspondent volume in carcass. Furthermore, LMVU was also useful in predicting the amounts of carcass tissues. It is possible to predict LM volume in the carcass using the LM volume measured in vivo by RTU. The amount of carcass tissues can be predicted by the LM volume measured in vivo by RTU.

The Effect of a Supragingival Plaque-Control Regimen on the Subgingival Microbiota in Smokers and Never-Smokers: Evaluation by Real-Time Polymerase Chain Reaction

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Anti-poliovirus activity of Baccharis dracunculifolia and propolis by cell viability determination and real-time PCR

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Conversion of web texts into Braille characters for real time reading through arrays of electromechanical actuators

This paper describes an electronic device conceived to convert common web texts into sequences of corresponding Braille signals, which are immediately reproduced onto an array ( keyboard) of electromechanical actuators. These actuators are reconfigurable in real time, displaying the Braille characters as matrices of points composed by small stems which can be lowered or raised according to the Braille code. The device, together with its conversion software package, can provide direct access to web texts in any personal computer, thus avoiding the use of complicated Braille printers.

Stochastic quantization of real-time thermal field theory

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Polymerase chain reaction and real-time PCR for diagnosing of Leishmania infantum chagasi in dogs

A importância do cão como reservatório de L. infantum chagasi no meio urbano tem estimulado a realização de inúmeros trabalhos de avaliação de técnicas de diagnóstico, uma vez que este procedimento, quando realizado corretamente, torna-se um importante passo na prevenção da doença em humanos. Dentre os métodos de diagnóstico, as técnicas moleculares têm adquirido destaque. Objetivou-se neste trabalho verificar o desempenho da Reação em Cadeia da Polimerase (PCR) e da PCR em tempo real (qPCR) para diagnóstico da Leishmaniose Visceral Canina (LVC) utilizando diferentes amostras biológicas. Para tanto foram utilizados 35 cães provenientes de uma área endêmica para LVC, onde foram utilizados para o diagnóstico molecular, aspirado de medula óssea, fragmentos de linfonodo e baço. Neste estudo a qPCR foi capaz de detectar um maior número de animais positivos quando comparada com a PCR. Já entre as diferentes amostras biológicas utilizadas não foi observada diferença significativa na detecção de DNA de L. infantumchagasi por meio da PCR e qPCR. Mesmo assim, considerando a facilidade de obtenção, o linfonodo pode ser considerada como a melhor amostra para diagnóstico molecular da infecção por L. infantum chagasi.

Quantitative real-time PCR identifies a critical region of deletion on 22q13 related to prognosis in oral cancer

Quantitative real time PCR was performed on genomic DNA from 40 primary oral carcinomas and the normal adjacent tissues. The target genes ECGFB, DIA1, BIK, and PDGFB and the microsatellite markers D22S274 and D22S277, mapped on 22q13, were selected according to our previous loss of heterozygosity findings in head and neck tumors. Quantitative PCR relies on the comparison of the amount of product generated from a target gene and that generated from a disomic reference gene (GAPDH-housekeeping gene). Reactions have been performed with normal control in triplicates, using the 7700 Sequence Detection System (PE Applied Biosystems). Losses in the sequences D22S274 (22q13.31) and in the DIA1 (22q13.2-13.31) gene were detected in 10 out of 40 cases (25%) each. Statistically significant correlations were observed for patients with relative copy number loss of the marker D22S274 and stages T3-T4 of disease (P=0.025), family history of cancer (P = 0.001), and death (P = 0.021). Relative copy number loss involving the DIA1 gene was correlated to family history of cancer (P<0.001), death (P=0.002), and consumption of alcohol (P=0.026). Log-rank test revealed a significant decrease in survival (P=0.0018) for patients with DIA1 gene loss. Relative copy number losses detected in these sequences may be related to disease progression and a worse prognosis in patients with oral cancer.

Progress on holographic techniques to measure real-time phase and amplitude gratings in photosensitive materials

In this paper we present and demonstrate a technique that allows simultaneous and independent measurement of small changes in the refractive index and the absorption coefficient produced in photosensitive materials during holographic exposure. The technique is based on the synchronous detection of two-wave mixing signals in both directions of the transmitted interfering beams. By processing both signals it is possible to separate the diffraction contributions of the refractive index from the absorption coefficient and simultaneously stabilize the incident fringe pattern. The demonstration of this technique is undertaken by following the temporal evolution of the phase and amplitude modulations in photoresist films. To check the ability of the technique to perform numeric evaluations, for a positive photoresist the changes in the optical constants were measured and compared with those obtained using independent methods.

Predicting beef carcass retail products of Mediterranean buffaloes by real-time ultrasound measures

Twenty eight Mediterranean buffaloes bulls were scanned with real-time ultrasound (RTU), slaughtered, and fabricated into retail cuts to determine the potential for ultrasound measures to predict carcass retail yield. Ultrasound measures of fat thickness, ribeye area and rump fat thickness were recorded three to five days prior to slaughter. Carcass measurements were taken, and one side of each carcass was fabricated into retail cuts. Stepwise regression analysis was used to compare possible models for prediction of either kilograms or percent retail product from carcass mesaurements and ultrasound measures. Results indicate that possible prediction models for percent or kilograms of retail products using RTU measures were similar in their predictive power and accuracy when compared to models derived from carcass measurements. Both fat thickness and ribeye area were over-predicted when measured ultrasonically compared to measurements taken on the carcass in the cooler. The mean absolute differences for both traits are larger than the mean differences, indicating that some images were interpreted to be larger and some smaller than actual carcass measurements. Ultrasound measurements of REA and FT had positive correlations with carcass measures of the same traits (r=.96 for REA and r=.99 for FT). Standard errors of prediction currently are being used as the standard to certify ultrasound technicians for accuracy. Regression equations using live weight (LW), rib eye area (REAU) and subcutaneous fat thickness (FTU) between 12(th) and 13 (th) ribs and also over the biceps femoris muscle (FTP8) by ultrasound explained 95% of the variation in the hot carcass weight when measure immediately before slaughter.

Periodontal status in smokers and never-smokers: Clinical findings and real-time polymerase chain reaction quantification of putative periodontal pathogens

Background: Smoking is a well-known risk factor for destructive periodontal disease, but its relationship with periodontal status and subgingival microbiota remains unclear. Inherent limitations of microbiological methods previously used may partly explain these mixed results, and real-time polymerase chain reaction (PCR) has been presented as a valid alternative. The aim of the present study was to investigate the clinical condition and microbiological profile of patients with chronic periodontitis as related to the habit of smoking.Methods: Fifty patients (33 to 59 years old), 25 smokers and 25 never-smokers, constituted the sample. The visible plaque index (VPI), gingival bleeding index (GBI), bleeding on probing (BOP), periodontal probing depth (PD), clinical attachment loss (CAL), and gingival crevicular fluid (GCF) volume were recorded. Real-time PCR quantified Porphyromonas gingivalis, Micromonas micros, Dialister pneumosintes, Actinobacillus actinomycetemcomitans and total bacteria in subgingival samples.Results: Smokers and never-smokers showed similar values for VPI, GBI, and BOP. Smokers had deeper PD in buccal/lingual sites and higher CAL independently of the tooth surface. The GCF volume was smaller in smokers, independent of the PD. Similar amounts of total bacteria and P. gingivalis were observed for both groups. Significantly higher numbers of D. pneumosintes and M. micros were present in smokers and associated with moderate and deep pockets. When heavy smokers were considered, higher counts of total bacteria, M. micros, and D. pneumosintes were observed.Conclusions: Smoking seems to have a detrimental impact on the periodontal status and microbiological profile of patients with periodontitis. Compared to never-smokers, smokers had deeper pockets, greater periodontal destruction, and higher counts of some putative periodontal pathogens.