991 resultados para Chromogenic In Situ Hybridization


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A sensitive and affordable approach is described for the in-situ measurement of ambient formaldehyde. Air is sampled around a 100 microliter aqueous drop containing 3-methyl-2-benzothiazoline hydrazone. After a desired period of sampling (typ. 5 min) and a waiting period of 10 min for the reaction to be completed, a second reagent (FeCl3) is added to the drop by means of a conjoined conduit. A blue product is formed and is read after an additional 10 min of reaction by a fiber-optic/light emitting diode based photodetector. A fresh drop is then formed and the process begins anew. As demonstrated here, the limit of detection is similar to 6.25 mu g m(-3) HCHO but can be significantly improved by using longer sampling times and a sampling rate higher than 100 mi min(-1) used in most of this work. This is the first example of a chromogenic drop sensor that utilizes sequential reagent addition.

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Members of the transforming growth factor-β family play critical roles in body patterning, in both vertebrates and invertebrates. One transforming growth factor-β-related gene, dbl-1, has been shown to regulate body length and male ray patterning in Caenorhabditis elegans. We screened arrayed cDNAs to identify downstream target genes for the DBL-1 signaling by using differential hybridization. C. elegans cDNAs representing 7,584 independent genes were arrayed on a nylon membrane at high density and hybridized with 33P-labeled DNA probes synthesized from the mRNAs of wild-type, dbl-1, sma-2, and lon-2 worms. Signals for all the spots representing hybridized DNA were quantified and compared among strains. The screening identified 22 and 2 clones, which were positively and negatively regulated, respectively, by the DBL-1 signal. Northern hybridization confirmed the expression profiles of most of the clones, indicating good reliability of the differential hybridization using arrayed cDNAs. In situ hybridization analysis revealed the spatial and temporal expression patterns of each clone and showed that at least four genes, including the gene for the type I receptor for DBL-1, sma-6, were transcriptionally regulated by the DBL-1 signal.

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Lobular carcinoma in situ was first described over 60 years ago. Despite the long history, it continues to pose significant difficulties in screening, diagnosis, management and treatment. This is partly due its multi-focal and bilateral presentation, an incomplete understanding of its biology and natural history and perpetuation of misconceptions gathered over the last decades. In this review, the working group on behalf of EUSOMA has attempted to summarise the current thinking and management of this interesting lesion. (c) 2006 Elsevier Ltd. All rights reserved.

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The morphological and chemical changes occurring during the thermal decomposition of weddelite, CaC2O4·2H2O, have been followed in real time in a heating stage attached to an Environmental Scanning Electron Microscope operating at a pressure of 2 Torr, with a heating rate of 10 °C/min and an equilibration time of approximately 10 min. The dehydration step around 120 °C and the loss of CO around 425 °C do not involve changes in morphology, but changes in the composition were observed. The final reaction of CaCO3 to CaO while evolving CO2 around 600 °C involved the formation of chains of very small oxide particles pseudomorphic to the original oxalate crystals. The change in chemical composition could only be observed after cooling the sample to 350 °C because of the effects of thermal radiation.

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Ceramic membranes were fabricated by in situ synthesis of alumina nanofibres in the pores of an alumina support as a separation layer, and exhibited a high permeation selectivity for bovine serum albumin relative to bovine hemoglobin (over 60 times) and can effectively retain DNA molecules at high fluxes.

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Using an in situ-generated calcium-based initiating species derived from pentaerythritol, the bulk synthesis of well-defined 4-arm star poly(L-lactide) oligomers has been studied in detail. The substitution of the traditional initiator, stannous octoate with calcium hydride allowed the synthesis of oligomers that had both low PDIs and a comparable number of polymeric arms (3.7 – 3.9) to oligomers of similar molecular weight. Investigations into the degree of control observed during the course of the polymerization found that the insolubility of pentaerythritol in molten L-lactide resulted in an uncontrolled polymerization only when the feed mole ratio of L-lactide to pentaerythritol was 13. At feed ratios of 40 and greater, a pseudo-living polymerization was observed. As part of this study, in situ FT-Raman spectroscopy was demonstrated to be a suitable method to monitor the kinetics of the ring-opening polymerization (ROP) of lactide. The advantages of using this technique rather than FT-IR-ATR and 1H NMR for monitoring L-lactide consumption during polymerization are discussed.

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It is known that adenosine 5'-triphosphate (ATP) is a cotransmitter in the heart. Additionally, ATP is released from ischemic and hypoxic myocytes. Therefore, cardiac-derived sources of ATP have the potential to modify cardiac function. ATP activates P2X(1-7) and P2Y(1-14) receptors; however, the presence of P2X and P2Y receptor subtypes in strategic cardiac locations such as the sinoatrial node has not been determined. An understanding of P2X and P2Y receptor localization would facilitate investigation of purine receptor function in the heart. Therefore, we used quantitative PCR and in situ hybridization to measure the expression of mRNA of all known purine receptors in rat left ventricle, right atrium and sinoatrial node (SAN), and human right atrium and SAN. Expression of mRNA for all the cloned P2 receptors was observed in the ventricles, atria, and SAN of the rat. However, their abundance varied in different regions of the heart. P2X(5) was the most abundant of the P2X receptors in all three regions of the rat heart. In rat left ventricle, P2Y(1), P2Y(2), and P2Y(14) mRNA levels were highest for P2Y receptors, while in right atrium and SAN, P2Y(2) and P2Y(14) levels were highest, respectively. We extended these studies to investigate P2X(4) receptor mRNA in heart from rats with coronary artery ligation-induced heart failure. P2X(4) receptor mRNA was upregulated by 93% in SAN (P < 0.05), while a trend towards an increase was also observed in the right atrium and left ventricle (not significant). Thus, P2X(4)-mediated effects might be modulated in heart failure. mRNA for P2X(4-7) and P2Y(1,2,4,6,12-14), but not P2X(2,3) and P2Y(11), was detected in human right atrium and SAN. In addition, mRNA for P2X(1) was detected in human SAN but not human right atrium. In human right atrium and SAN, P2X(4) and P2X(7) mRNA was the highest for P2X receptors. P2Y(1) and P2Y(2) mRNA were the most abundant for P2Y receptors in the right atrium, while P2Y(1), P2Y(2), and P2Y(14) were the most abundant P2Y receptor subtypes in human SAN. This study shows a widespread distribution of P2 receptor mRNA in rat heart tissues but a more restricted presence and distribution of P2 receptor mRNA in human atrium and SAN. This study provides further direction for the elucidation of P2 receptor modulation of heart rate and contractility.

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Aim: To measure the influence of spherical intraocular lens implantation and conventional myopic laser in situ keratomileusis on peripheral ocular aberrations. Setting: Visual & Ophthalmic Optics Laboratory, School of Optometry & Institute of Health and Biomedical Innovation, Queensland University of Technology, Brisbane, Australia. Methods: Peripheral aberrations were measured using a modified commercial Hartmann-Shack aberrometer across 42° x 32° of the central visual field in 6 subjects after spherical intraocular lens (IOL) implantation and in 6 subjects after conventional laser in situ keratomileusis (LASIK) for myopia. The results were compared with those of age matched emmetropic and myopic control groups. Results: The IOL group showed a greater rate of quadratic change of spherical equivalent refraction across the visual field, higher spherical aberration, and greater rates of change of higher-order root-mean-square aberrations and total root-mean-square aberrations across the visual field than its emmetropic control group. However, coma trends were similar for the two groups. The LASIK group had a greater rate of quadratic change of spherical equivalent refraction across the visual field, higher spherical aberration, the opposite trend in coma across the field, and greater higher-order root-mean-square aberrations and total root-mean-square aberrations than its myopic control group. Conclusion: Spherical IOL implantation and conventional myopia LASIK increase ocular peripheral aberrations. They cause considerable increase in spherical aberration across the visual field. LASIK reverses the sign of the rate of change in coma across the field relative to that of the other groups. Keywords: refractive surgery, LASIK, IOL implantation, aberrations, peripheral aberrations

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This paper examines the ground-water flow problem associated with the injection and recovery of certain corrosive fluids into mineral bearing rock. The aim is to dissolve the minerals in situ, and then recover them in solution. In general, it is not possible to recover all the injected fluid, which is of concern economically and environmentally. However, a new strategy is proposed here, that allows all the leaching fluid to be recovered. A mathematical model of the situation is solved approximately using an asymptotic solution, and exactly using a boundary integral approach. Solutions are shown for two-dimensional flow, which is of some practical interest as it is achievable in old mine tunnels, for example.

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Gaze and movement behaviors of association football goalkeepers were compared under two video simulation conditions (i.e., verbal and joystick movement responses) and three in situ conditions (i.e., verbal, simplified body movement, and interceptive response). The results showed that the goalkeepers spent more time fixating on information from the penalty kick taker’s movements than ball location for all perceptual judgment conditions involving limited movement (i.e., verbal responses, joystick movement, and simplified body movement). In contrast, an equivalent amount of time was spent fixating on the penalty taker’s relative motions and the ball location for the in situ interception condition, which required the goalkeepers to attempt to make penalty saves. The data suggest that gaze and movement behaviors function differently, depending on the experimental task constraints selected for empirical investigations. These findings highlight the need for research on perceptual— motor behaviors to be conducted in representative experimental conditions to allow appropriate generalization of conclusions to performance environments.