Validation of dot blot hybridization and denaturing high performance liquid chromatography as reliable methods for TP53 codon 72 genotyping in molecular epidemiologic studies

Background: Mutations in TP53 are common events during carcinogenesis. In addition to gene mutations, several reports have focused on TP53 polymorphisms as risk factors for malignant disease. Many studies have highlighted that the status of the TP53 codon 72 polymorphism could influence cancer susceptibility. However, the results have been inconsistent and various methodological features can contribute to departures from Hardy-Weinberg equilibrium, a condition that may influence the disease risk estimates. The most widely accepted method of detecting genotyping error is to confirm genotypes by sequencing and/or via a separate method. Results: We developed two new genotyping methods for TP53 codon 72 polymorphism detection: Denaturing High Performance Liquid Chromatography (DHPLC) and Dot Blot hybridization. These methods were compared with Restriction Fragment Length Polymorphism (RFLP) using two different restriction enzymes. We observed high agreement among all methodologies assayed. Dot-blot hybridization and DHPLC results were more highly concordant with each other than when either of these methods was compared with RFLP. Conclusions: Although variations may occur, our results indicate that DHPLC and Dot Blot hybridization can be used as reliable screening methods for TP53 codon 72 polymorphism detection, especially in molecular epidemiologic studies, where high throughput methodologies are required.

Rapid and sensitive measurements of nitrate ester explosives using microchip electrophoresis with electrochemical detection

This article describes an effective microchip protocol based on electrophoretic-separation and electrochemical detection for highly sensitive and rapid measurements of nitrate ester explosives, including ethylene glycol dinitrate (EGDN), pentaerythritol tetranitrate (PETN), propylene glycol dinitrate (PGDN) and glyceryl trinitrate (nitroglycerin, NG). Factors influencing the separation and detection processes were examined and optimized. Under the optimal separation conditions obtained using a 15 mM borate buffer (pH 9.2) containing 20 mM SDS, and applying a separation voltage of 1500 V, the four nitrate ester explosives were separated within less than 3 min. The glassy-carbon amperometric detector (operated at -0.9 V vs. Ag/AgCl) offers convenient cathodic detection down to the picogram level, with detection limits of 0.5 ppm and 0.3 ppm for PGDN and for NG, respectively, along with good repeatability (RSD of 1.8-2.3%; n = 6) and linearity (over the 10-60 ppm range). Such effective microchip operation offers great promise for field screening of nitrate ester explosives and for supporting various counter-terrorism surveillance activities.

Evaluation of grinding methods for pellets preparation aiming at the analysis of plant materials by laser induced breakdown spectrometry

It has been demonstrated that laser induced breakdown spectrometry (LIBS) can be used as an alternative method for the determination of macro (P, K. Ca, Mg) and micronutrients (B, Fe, Cu, Mn, Zn) in pellets of plant materials. However, information is required regarding the sample preparation for plant analysis by LIBS. In this work, methods involving cryogenic grinding and planetary ball milling were evaluated for leaves comminution before pellets preparation. The particle sizes were associated to chemical sample properties such as fiber and cellulose contents, as well as to pellets porosity and density. The pellets were ablated at 30 different sites by applying 25 laser pulses per site (Nd:YAG@1064 nm, 5 ns, 10 Hz, 25J cm(-2)). The plasma emission collected by lenses was directed through an optical fiber towards a high resolution echelle spectrometer equipped with an ICCD. Delay time and integration time gate were fixed at 2.0 and 4.5 mu s, respectively. Experiments carried out with pellets of sugarcane, orange tree and soy leaves showed a significant effect of the plant species for choosing the most appropriate grinding conditions. By using ball milling with agate materials, 20 min grinding for orange tree and soy, and 60 min for sugarcane leaves led to particle size distributions generally lower than 75 mu m. Cryogenic grinding yielded similar particle size distributions after 10 min for orange tree, 20 min for soy and 30 min for sugarcane leaves. There was up to 50% emission signal enhancement on LIBS measurements for most elements by improving particle size distribution and consequently the pellet porosity. (C) 2011 Elsevier B.V. All rights reserved.

Coexistence of forest and savanna in an Amazonian area from a geological perspective

Question: How can the coexistence of savanna and forest in Amazonian areas with relatively uniform climates be explained? Location: Eastern Marajo Island, northeast Amazonia, Brazil. Methods: The study integrated floristic analysis, terrain morphology, sedimentology and delta(13)C of soil organic matter. Floristic analysis involved rapid ecological assessment of 33 sites, determination of occurrence, specific richness, hierarchical distribution and matrix of floristic similarity between paired vegetation types. Terrain characterization was based on analysis of Landsat images using 4(R), 5(G) and 7(B) composition and digital elevation model (DEM). Sedimentology involved field descriptions of surface and core sediments. Finally, radiocarbon dating and analysis of delta(13)C of soil profile organic matter and natural ecotone forest-savanna was undertaken. Results: Slight tectonic subsidence in eastern Marajo Island favours seasonal flooding, making it unsuitable for forest growth. However, this area displays slightly convex-up, sinuous morphologies related to paleochannels, covered by forest. Terra-firme lowland forests are expanding from west to east, preferentially occupying paleochannels and replacing savanna. Slack, running water during channel abandonment leads to disappearance of varzea/gallery forest at channel margins. Long-abandoned channels sustain continuous terra-firme forests, because of longer times for more species to establish. Recently abandoned channels have had less time to become sites for widespread tree development, and are either not vegetated or covered by savanna. Conclusion: Landforms in eastern Marajo Island reflect changes in the physical environment due to reactivation of tectonic faults during the latest Quaternary. This promoted a dynamic history of channel abandonment, which controlled a set of interrelated parameters (soil type, topography, hydrology) that determined species location. Inclusion of a geological perspective for paleoenvironmental reconstruction can increase understanding of plant distribution in Amazonia.

Zone trapping/merging zones in flow analysis: A novel approach for rapid assays involving relatively slow chemical reactions

A novel strategy for accomplishing zone trapping in flow analysis is proposed. The sample and the reagent solutions are simultaneously inserted into convergent carrier streams and the established zones merge together before reaching the detector, where the most concentrated portion of the entire sample zone is trapped. The main characteristics, potentialities and limitations of the strategy were critically evaluated in relation to an analogous flow system with zone stopping. When applied to the spectrophotometric determination of nitrite in river waters, the main figures of merit were maintained, exception made for the sampling frequency which was calculated as 189h(-1), about 32% higher relatively to the analogous system with zone stopping. The sample inserted volume can be increased up to 1.0 mL without affecting sampling frequency and no problems with pump heating or malfunctions were noted after 8-h operation of the system. In contrast to zone stopping, only a small portion of the sample zone is halted with zone trapping, leading to these beneficial effects. (C) 2011 Elsevier B.V. All rights reserved.

Methods of imagetic information representation

The aim of this paper is to highlight some of the methods of imagetic information representation, reviewing the literature of the area and proposing a model of methodology adapted to Brazilian museums. An elaboration of a methodology of imagetic information representation is developed based on Brazilian characteristics of information treatment in order to adapt it to museums. Finally, spreadsheets that show this methodology are presented.

Methods of quantification of training load in exercise performed at maximal lactate steady state

The aim of the present study was to compare and correlate training impulse (TRIMP) estimates proposed by Banister (TRIMP(Banister)), Stagno (TRIMP(Stagno)) and Manzi (TRIMP(Manzi)). The subjects were submitted to an incremental test on cycle ergometer with heart rate and blood lactate concentration measurements. In the second occasion, they performed 30 min. of exercise at the intensity corresponding to maximal lactate steady state, and TRIMP(Banister), TRIMP(Stagno) and TRIMP(Manzi) were calculated. The mean values of TRIMP(Banister) (56.5 +/- 8.2 u.a.) and TRIMP(Stagno) (51.2 +/- 12.4 u.a.) were not different (P > 0.05) and were highly correlated (r = 0.90). Besides this, they presented a good agreement level, which means low bias and relatively narrow limits of agreement. On the other hand, despite highly correlated (r = 0.93), TRIMP(Stagno) and TRIMP(Manzi) (73.4 +/- 17.6 u.a.) were different (P < 0.05), with low agreement level. The TRIMP(Banister) e TRIMP(Manzi) estimates were not different (P = 0.06) and were highly correlated (r = 0.82), but showed low agreement level. Thus, we concluded that the investigated TRIMP methods are not equivalent. In practical terms, it seems prudent monitor the training process assuming only one of the estimates.

Testing motor fitness in karate

Background and Study Aim: Evaluation of sport skills test can be very useful tool for coach practice. The aim of the present paper was: (a) to evaluate the reliability and accuracy of the Specific Physical Fitness Tests (SPFT) (b) to review the results of karate athletes who represent different weight categories, and who are at different stages of schooling; (c) to establish grading criteria of physical fitness preparation. Material/Methods: The reseach was conducted among 219 Kyokushin karate players, whose profiles were presented as (chi) over bar +/- SD and their main characteristics were the following: age 26.8 +/- 4.67 (19-39) years, body mass 75.2 +/- 8.35 (50-97) kg and body height 176.4 +/- 5.67 (160-196) cm. The value of the BMI amounted to 24.1 +/- 2.17 (17.9-29.4) kg/m(2). All the subjects of the research had training experience of 10.5 +/- 3.71 (4-20) years and their degree of proficiency ranged from 4(th) kyu to 3(rd) dan. The physical fitness trials proposed by Story (1989) included: hip turning speed, speed punches, flexibility, rapid kicks, agility, and evasion actions. It was supplemented by a test of local strength endurance, composing a battery of the SPFT, which was implemented by first of the authors between 1991 and 2006. Results: SPFT is characterized by high reliability and it can be used to diagnose the physical fitness preparation and monitor the individual results of training. It discriminates accurately competitors with different sports level and it is characterized by very high accuracy, it is correlated with the test results of motor general physical fitness abilities and coordination abilities as well as it is connected with the somatic build of the athlete. The performance classification table was developed on the basis of our research. Discussion: Results obtained in SPFT were shortly discussed. Conclusions: The collected results of our research allowed us to come to, the conclusion: The table can be applied not only to assess karate fighters, but also adepts in taekwondo, kick-boxing, ju-jitsu, hapkido or other mixed martial arts.

Propofol-induced sleep: Polysomnographic evaluation of patients with obstructive sleep apnea and controls

OBJECTIVE: The localization of upper airway obstruction in patients with obstructive sleep apnea (OSA) may optimize treatment. Nasoendoscopy during propofol sedation allows such an evaluation, but the effect of this drug on respiratory patterns and muscle relaxation is unknown. The objective of the present study was to determine through polysomnography whether propofol would change sleep parameters. STUDY DESIGN: Prospective study of subjects submitted to polysomnography under sedation with propofol. SETTING: Tertiary referral center. SUBJECTS AND METHODS: Fifteen non-obese subjects (4 controls/11 OSA patients) were submitted to two diurnal polysomnograms (90-120 minutes of sleep), with and without the use of propofol. The parameters presence of snoring, apnea-hypopnea index (AHI), oxygen desaturation, and sleep architecture were compared. RESULTS: The use of propofol did not induce snoring in the control subjects, whereas 100 percent of the OSA patients snored. AHI and mean oxygen saturation (SaO(2)) did not differ significantly between examinations with and without sedation. However, minimum SaO(2) differed significantly (P < 0.05) with sedation, being lower during propofol sedation. Propofol also significantly changed the sleep architecture, with a significant increase in N3 sleep (P < 0.005) and total abolishment of rapid eye movement sleep (P < 0.0005) during propofol sedation. CONCLUSIONS: These preliminary results allow us to infer that sedation with propofol changes sleep architecture but permits respiratory evaluation, because the main respiratory parameters evaluated in OSA are maintained. These preliminary results support the view that nasoendoscopy under propofol sedation is a promising examination for management of this disease. (C) 2010 American Academy of Otolaryngology-Head and Neck Surgery Foundation. All rights reserved.

Rapid weight loss followed by recovery time does not affect judo-related performance

In this study, we investigated the effects of rapid weight loss followed by a 4-h recovery on judo-related performance. Seven weight-cycler athletes were assigned to a weight loss group (5% body weight reduction by self-selected regime) and seven non-weight-cyclers to a control group (no weight reduction). Body composition, performance, glucose, and lactate were assessed before and after weight reduction (5-7 days apart; control group kept weight stable). The weight loss group had 4 h to re-feed and rehydrate after the weigh-in. Food intake was recorded during the weight loss period and recovery after the weigh-in. Performance was evaluated through a specific judo exercise, followed by a 5-min judo combat and by three bouts of the Wingate test. Both groups significantly improved performance after the weight loss period. No interaction effects were observed. The energy and macronutrient intake of the weight loss group were significantly lower than for the control group. The weight loss group consumed large amounts of food and carbohydrate during the 4-h recovery period. No changes were observed in lactate concentration, but a significant decrease in glucose during rest was observed in the weight loss group. In conclusion, rapid weight loss did not affect judo-related performance in experienced weight-cyclers when the athletes had 4 h to recover. These results should not be extrapolated to inexperienced weight-cyclers.

Estimation of hexenuronic acids and kappa number in kraft pulps of Eucalyptus globulus by Fourier transform near infrared spectroscopy and multivariate analysis

Fourier transform near infrared (FT-NIR) spectroscopy was evaluated as an analytical too[ for monitoring residual Lignin, kappa number and hexenuronic acids (HexA) content in kraft pulps of Eucalyptus globulus. Sets of pulp samples were prepared under different cooking conditions to obtain a wide range of compound concentrations that were characterised by conventional wet chemistry analytical methods. The sample group was also analysed using FT-NIR spectroscopy in order to establish prediction models for the pulp characteristics. Several models were applied to correlate chemical composition in samples with the NIR spectral data by means of PCR or PLS algorithms. Calibration curves were built by using all the spectral data or selected regions. Best calibration models for the quantification of lignin, kappa and HexA were proposed presenting R-2 values of 0.99. Calibration models were used to predict pulp titers of 20 external samples in a validation set. The lignin concentration and kappa number in the range of 1.4-18% and 8-62, respectively, were predicted fairly accurately (standard error of prediction, SEP 1.1% for lignin and 2.9 for kappa). The HexA concentration (range of 5-71 mmol kg(-1) pulp) was more difficult to predict and the SEP was 7.0 mmol kg(-1) pulp in a model of HexA quantified by an ultraviolet (UV) technique and 6.1 mmol kg(-1) pulp in a model of HexA quantified by anion-exchange chromatography (AEC). Even in wet chemical procedures used for HexA determination, there is no good agreement between methods as demonstrated by the UV and AEC methods described in the present work. NIR spectroscopy did provide a rapid estimate of HexA content in kraft pulps prepared in routine cooking experiments.

Preparation and characterization of Mo/W bimetallic carbides by using different synthesis methods

Molybdenum and tungsten bimetallic oxides were synthetized according to the following methods: Pechini, coprecipitation and solid state reaction (SSR). After the characterization, those solids were carbureted at programmed temperature. The carburation process was monitored by checking the consumption of carburant hydrocarbon and CO produced. The monitoring process permits to avoid or to diminish the formation of pirolytic carbon.

Rapid solidification and phase stability evaluation of Ti-Si-B alloys

Ti-rich Ti-Si-B alloys can be considered for structural applications at high temperatures (max. 700 degrees C), however, phase equilibria data is reported only for T = 1250 degrees C. Thus, in this work the phase stability of this system has been evaluated at 700 degrees C. In order to attain equilibrium conditions in shorter time, rapid solidified samples have been prepared and carefully characterized. The microstructural characterization of the produced materials were based on X-ray diffraction (XRD), scanning electron microscopy (SEM-BSE), high resolution transmission electron microscopy (HRTEM), High Temperature X-ray diffraction with Synchrotron radiation (XRDSR) and Differential Scanning Calorimetry (DSC). Amorphous and amorphous with embedded nanocrystals have been observed after rapid solidification from specific alloy compositions. The values of the crystallization temperature (Tx) of the alloys were in the 509-647 degrees C temperature range. After Differential Scanning Calorimetry and High Temperature X-ray Diffraction with Synchrotron radiation, the alloys showed crystalline and basically formed by two or three of the following phases: alpha Ti, Ti(6)Si(2)B; Ti(5)Si(3); Ti(3)Si and TiB. It has been shown the stability of the Ti(3)Si and Ti(6)Si(2)B phases at 700 degrees C and the proposition of an isothermal section at this temperature. (C) 2011 Elsevier B.V. All rights reserved.

Ecodesign methods focused on remanufacturing

Understanding the product`s `end-of-life` is important to reduce the environmental impact of the products` final disposal. When the initial stages of product development consider end-of-life aspects, which can be established by ecodesign (a proactive approach of environmental management that aims to reduce the total environmental impact of products), it becomes easier to close the loop of materials. The `end-of-life` ecodesign methods generally include more than one `end-of-life` strategy. Since product complexity varies substantially, some components, systems or sub-systems are easier to be recycled, reused or remanufactured than others. Remanufacture is an effective way to maintain products in a closed-loop, reducing both environmental impacts and costs of the manufacturing processes. This paper presents some ecodesign methods focused on the integration of different `end-of-life` strategies, with special attention to remanufacturing, given its increasing importance in the international scenario to reduce the life cycle impacts of products. (C) 2009 Elsevier Ltd. All rights reserved.

A Study on Unconditionally Stable FDTD Methods for the Modeling of Metamaterials

We assess the performance of three unconditionally stable finite-difference time-domain (FDTD) methods for the modeling of doubly dispersive metamaterials: 1) locally one-dimensional FDTD; 2) locally one-dimensional FDTD with Strang splitting; and (3) alternating direction implicit FDTD. We use both double-negative media and zero-index media as benchmarks.