Evaluation of HER2, p53, bcl-2, topoisomerase II-alpha, heat shock proteins 27 and 70 in primary breast cancer and metastatic ipsilateral axillary lymph nodes.

BACKGROUND: Breast cancer is a heterogeneous disease. Predictive biological markers (BM) of responsiveness to therapy need to be identified. Evaluation of BM is mainly done at the primary site. However, in the adjuvant therapy of breast cancer, the main goal is control of micrometastases. It is still unknown whether heterogeneity in the expression of BM between the primary site and its micrometastases exists. OBJECTIVE: To evaluate the expression of some BM with potential predictive value from the primary breast cancer site and metastatic ipsilateral axillary lymph nodes. PATIENTS AND METHODS: Focality (percentage of positive cells) and intensity staining scores were evaluated for each marker. Freshly cut sections (4 microm) from embedded blocks of breast cancer fixed in formalin or bouin were put onto superfrost slides (Menzel-Gläser). Protein expression was evaluated immunohistochemically (IHC) using monoclonal antibodies against: topo II-alpha (clone KiS1, 1 microg/ml, Roche) with a trypsine pre-treatment (P); HSP27 (clone G3.1, 1/60, Biogenex), HSP70 (clone BRM.22, 1/80, Biogenex) and HER2 (clone CB11, 1/40, Novocastra; without P); p53 (clone D07, 1/750, Dako) and bcl-2 (clone 124, 1/60, Dako) with citrate buffer as P. RESULTS: Overall, the percentage of discordant marker status in the primary tumour and its metastatic lymph nodes was 2% for HER2, 6% for p53, 15% for bcl-2, 19% for topoisomerase II-alpha, 24% for HSP27 and 30% for HSP70. For the subgroup of patients with positive BM in the primary tumour, the percentage of discordance was 6% for HER2, 7% for p53, 14% for bcl-2, 19% for HSP70, 21% for topoisomerase II-alpha and 36% for HSP27. For the subgroup of patients with positive BM in the lymph nodes, the percentage of discordance was 9% for bcl-2, 15% for HER2 and p53, 21% for topoisomerase II-alpha, 22% for HSP27 and 25% for HSP70. CONCLUSIONS: 1) No biological marker had 100% concordant results. 2) Although some discordant cases might be explained by the limitations of the IHC technique, future studies aiming to evaluate the predictive value of BM in the adjuvant therapy of breast cancer should take into account a possible difference in BM expression between the primary and the metastatic sites.

On the adsorption of hexaammineruthenium (III) at anionic self-assembled monolayers

The binding of the electroactive hexaammineruthenium (III) complex ions to anionic self-assembled monolayers (SAMs) has been investigated by means of chronocoulometry and ac voltammetry. From chronocoulometric data recorded in 10-2 M LiClO4 containing different [Ru(NH3)6]3+ concentrations, we have established the adsorption isotherm of [Ru(NH3)6]3+ on a compact monolayer of 2-mercaptobenzimidazole-5-sulfonate (MBIS) self-assembled on Au(1 1 1). The data were satisfactorily fitted to the linearized Langmuir adsorption isotherm and a binding constant of 4.0 (±0.4) × 106 M-1 has been determined. The electrostatic binding of [Ru(NH3)6]3+ to a dilute PNA-DNA monolayer formed after hybridization on a PNA-modified gold electrode by self-assembly from a mixed solution of mercaptobutan-1-ol and PNA oligonucleotides has been studied by ac voltammetry. The admittance of the PNA-modified electrode after hybridization with complementary DNA was measured in 0.01 M Tris-HCl buffer containing different [Ru(NH3)6]3+ concentrations. Based on these data, a binding constant of [Ru(NH3)6]3+ to the surface-confined PNA-DNA duplex was derived from the Langmuir isotherm and amounts to 2.9 (±0.3) × 105 M-1. As the interactions between [Ru(NH3)6]3+ and the immobilized PNA-DNA hybrids on the gold surface are essentially electrostatic, the adsorption of the highly charged cationic redox complex at low concentrations to the negatively charged PNA-DNA modified surface is in large competition with other monovalent cations present in the electrolyte at higher concentrations. The influence of competing sodium cations was thus studied by adding different NaCl concentrations in the 0.01 M Tris-HCl electrolyte. © 2008 Elsevier Ltd. All rights reserved.

Membrane binding of plasmid DNA and endocytic pathways are involved in electrotransfection of mammalian cells.

Electric field mediated gene delivery or electrotransfection is a widely used method in various studies ranging from basic cell biology research to clinical gene therapy. Yet, mechanisms of electrotransfection are still controversial. To this end, we investigated the dependence of electrotransfection efficiency (eTE) on binding of plasmid DNA (pDNA) to plasma membrane and how treatment of cells with three endocytic inhibitors (chlorpromazine, genistein, dynasore) or silencing of dynamin expression with specific, small interfering RNA (siRNA) would affect the eTE. Our data demonstrated that the presence of divalent cations (Ca(2+) and Mg(2+)) in electrotransfection buffer enhanced pDNA adsorption to cell membrane and consequently, this enhanced adsorption led to an increase in eTE, up to a certain threshold concentration for each cation. Trypsin treatment of cells at 10 min post electrotransfection stripped off membrane-bound pDNA and resulted in a significant reduction in eTE, indicating that the time period for complete cellular uptake of pDNA (between 10 and 40 min) far exceeded the lifetime of electric field-induced transient pores (∼10 msec) in the cell membrane. Furthermore, treatment of cells with the siRNA and all three pharmacological inhibitors yielded substantial and statistically significant reductions in the eTE. These findings suggest that electrotransfection depends on two mechanisms: (i) binding of pDNA to cell membrane and (ii) endocytosis of membrane-bound pDNA.

Reexamining the science of marine protected areas: Linking knowledge to action

Marine protected areas (MPAs) are often implemented to conserve or restore species, fisheries, habitats, ecosystems, and ecological functions and services; buffer against the ecological effects of climate change; and alleviate poverty in coastal communities. Scientific research provides valuable insights into the social and ecological impacts of MPAs, as well as the factors that shape these impacts, providing useful guidance or "rules of thumb" for science-based MPA policy. Both ecological and social factors foster effective MPAs, including substantial coverage of representative habitats and oceanographic conditions; diverse size and spacing; protection of habitat bottlenecks; participatory decisionmaking arrangements; bounded and contextually appropriate resource use rights; active and accountable monitoring and enforcement systems; and accessible conflict resolution mechanisms. For MPAs to realize their full potential as a tool for ocean governance, further advances in policy-relevant MPA science are required. These research frontiers include MPA impacts on nontarget and wide-ranging species and habitats; impacts beyond MPA boundaries, on ecosystem services, and on resource-dependent human populations, as well as potential scale mismatches of ecosystem service flows. Explicitly treating MPAs as "policy experiments" and employing the tools of impact evaluation holds particular promise as a way for policy-relevant science to inform and advance science-based MPA policy. © 2011 Wiley Periodicals, Inc.

Community Vulnerability to Malaria in Madre de Dios, Peru

Construction of a highway and artisanal gold mining have contributed to population and land use changes within the department of Madre de Dios, Peru. Such changes are expected to alter malaria rates due to impacts on vector habitat and human exposure. Vulnerability, as defined by the possibility of bereavement of a physical good or abstract state, is useful for understanding which communities are most likely to be adversely impacted by hazards such as malaria. A model defining susceptibility (SUS) and lack of resilience (LOR) was used to create an index of vulnerability to malaria for 40 communities in Madre de Dios. Indicators of SUS and LOR were developed from household and community data and combined into a final vulnerability index score. Vulnerability scores ranged between 0.13 and 0.31 with a mean of 0.21. Communities were grouped according to standard deviations from the mean. The most vulnerable communities (>1.5 standard deviations from mean) were located in the southern portion of the study area. When the dimension scores were compared for all communities, scores were generally higher in the susceptibility dimension than in the lack of resilience dimension. Examination of the indicator scores of individual communities revealed that drivers of vulnerability vary across the department. Therefore, targeted interventions addressing specific aspects of vulnerability may be useful. Finally, a predicted vulnerability surface was created for a 10 km buffer surrounding the Interoceanic Highway in Madre de Dios.

Recognition of Planar Segments in Point Cloud Based on Wavelet Transform

© 2005-2012 IEEE.Within industrial automation systems, three-dimensional (3-D) vision provides very useful feedback information in autonomous operation of various manufacturing equipment (e.g., industrial robots, material handling devices, assembly systems, and machine tools). The hardware performance in contemporary 3-D scanning devices is suitable for online utilization. However, the bottleneck is the lack of real-time algorithms for recognition of geometric primitives (e.g., planes and natural quadrics) from a scanned point cloud. One of the most important and the most frequent geometric primitive in various engineering tasks is plane. In this paper, we propose a new fast one-pass algorithm for recognition (segmentation and fitting) of planar segments from a point cloud. To effectively segment planar regions, we exploit the orthonormality of certain wavelets to polynomial function, as well as their sensitivity to abrupt changes. After segmentation of planar regions, we estimate the parameters of corresponding planes using standard fitting procedures. For point cloud structuring, a z-buffer algorithm with mesh triangles representation in barycentric coordinates is employed. The proposed recognition method is tested and experimentally validated in several real-world case studies.

Elucidating the Molecular Composition of Cartilage by Proteomics.

Articular cartilage consists of chondrocytes and two major components, a collagen-rich framework and highly abundant proteoglycans. Most prior studies defining the zonal distribution of cartilage have extracted proteins with guanidine-HCl. However, an unextracted collagen-rich residual is left after extraction. In addition, the high abundance of anionic polysaccharide molecules extracted from cartilage adversely affects the chromatographic separation. In this study, we established a method for removing chondrocytes from cartilage sections with minimal extracellular matrix protein loss. The addition of surfactant to guanidine-HCl extraction buffer improved protein solubility. Ultrafiltration removed interference from polysaccharides and salts. Almost four-times more collagen peptides were extracted by the in situ trypsin digestion method. However, as expected, proteoglycans were more abundant within the guanidine-HCl extraction. These different methods were used to extract cartilage sections from different cartilage layers (superficial, intermediate, and deep), joint types (knee and hip), and disease states (healthy and osteoarthritic), and the extractions were evaluated by quantitative and qualitative proteomic analyses. The results of this study led to the identifications of the potential biomarkers of osteoarthritis (OA), OA progression, and the joint specific biomarkers.

Polymeric photosensitizer-embedded self-expanding metal stent for repeatable endoscopic photodynamic therapy of cholangiocarcinoma

Photodynamic therapy (PDT) is a new therapeutic approach for the palliative treatment of malignant bile duct obstruction. In this study, we designed photosensitizer-embedded self-expanding nonvascular metal stent (PDT-stent) which allows repeatable photodynamic treatment of cholangiocarcinoma without systemic injection of photosensitizer. Polymeric photosensitizer (pullulan acetate-conjugated pheophorbide A; PPA) was incorporated in self-expanding nonvascular metal stent. Residence of PPA in the stent was estimated in buffer solution and subcutaneous implantation on mouse. Photodynamic activity of PDT-stent was evaluated through laserexposure on stent-layered tumor cell lines, HCT-116 tumor-xenograft mouse models and endoscopic intervention of PDT-stent on bile duct of mini pigs. Photo-fluorescence imaging of the PDT-stent demonstrated homogeneous embedding of polymeric Pheo-A (PPA) on stent membrane. PDT-stent sustained its photodynamic activities at least for 2 month. And which implies repeatable endoscopic PDT is possible after stent emplacement. The PDT-stent after light exposure successfully generated cytotoxic singlet oxygen in the surrounding tissues, inducing apoptotic degradation of tumor cells and regression of xenograft tumors on mouse models. Endoscopic biliary in-stent photodynamic treatments on minipigs also suggested the potential efficacy of PDT-stent on cholangiocarcinoma. In vivo and in vitro studies revealed our PDT-stent, allows repeatable endoscopic biliary PDT, has the potential for the combination therapy (stent plus PDT) of cholangiocarcinoma. © 2014 Elsevier Ltd.

Photophysics of bifunctional Ru(II) complexes bearing an aminoquinoline organic unit. Potential new photoprobes and photoreagents of DNA

[Ru(BPY)2POQ-Nmet]2+ and [Ru(TAP)2POQ-Nmet]2+ (1 and 3) are bifunctional complexes composed of a metallic unit linked by a flexible chain to an organic unit. They have been prepared as photoprobes or photoreagents of DNA. In this work, the spectroscopic properties of these bifunctional complexes in the absence of DNA are compared with those of the monofunctional analogues [Ru(BPY)2Phen]2+, [Ru-(BPY)2acPhen]2+, [Ru(TAP)2Phen]2+, and [Ru(TAP)2acPhen]2+ (2 and 4). The electrospray mass spectrometry and absorption data show that the quinoline moiety exists in the protonated and nonprotonated form. Although the bifunctional complex containing 2,2′-bipyridine (BPY) ligands exhibits photophysical properties similar to those of the monofunctional compounds, the bifunctional complex with 1,4,5,8-tetraazaphenanthrene (TAP) ligands behaves quite differently. It has weaker relative emission quantum yields and shorter luminescence lifetimes than the monofunctional TAP analogue when the quinoline unit is nonprotonated. This indicates an efficient intramolecular quenching of the 3MLCT (metal to ligand charge transfer) excited state of the TAP metallic moiety. When the organic unit is protonated, there is no internal quenching. In organic solvent, the nonquenched excited metallic unit (bearing a protonated quinoline) and the quenched one (bearing a nonprotonated organic unit) are in slow equilibrium as compared to the lifetime of the two emitters. In aqueous solution this equilibrium is faster and is catalysed by the presence of phosphate buffer. Flash photolysis experiments suggest that the intramolecular quenching process originates from a photoinduced electron transfer from the nonprotonated quinoline to the excited Ru(TAP)2 2+ moiety.

Relaciones suelo planta de crisantemos (Dendranthema grandiflora) en maceta

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Indices de sorción de fósforo en algunos molisoles de la pradera pampeana

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Distintas fracciones de fósforo en suelos del norte de Entre Ríos

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Rutinas de fertilización de plantas ornamentales anuales en contenedores pequeños

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Performance evaluation of the register insertion protocol for voice-data integration

The performance of the register insertion protocol for mixed voice-data traffic is investigated by simulation. The simulation model incorporates a common insertion buffer for station and ring packets. Bandwidth allocation is achieved by imposing a queue limit at each node. A simple priority scheme is introduced by allowing the queue limit to vary from node to node. This enables voice traffic to be given priority over data. The effect on performance of various operational and design parameters such as ratio of voice to data traffic, queue limit and voice packet size is investigated. Comparisons are made where possible with related work on other protocols proposed for voice-data integration. The main conclusions are: (a) there is a general degradation of performance as the ratio of voice traffic to data traffic increases, (b) substantial improvement in performance can be achieved by restricting the queue length at data nodes and (c) for a given ring utilisation, smaller voice packets result in lower delays for both voice and data traffic.

Impacts of wildlife on household food security and income in northeastern Tanzania

We evaluated the impacts of wildlife on household food security and income in three semi-arid villages adjacent to Lake Manyara National Park (LMNP) and Mkomazi Game Reserve (MGR) in Northeastern Tanzania. Survey data were collected using both household interviews and human-wildlife conflict related archive information from the village government offices. Crop destruction by wildlife influenced both household food security and cash income. Crop damage to households was, on average, 0.08 ton/annum, equivalent to two months household loss of food and reduced household cash income by 1.3%. A combination of measures is proposed as incentives for conservation. These include provision of economic incentives, soft loans to initiate non-farm (e.g., ecotourism, business enterprises) projects to ease dependency on natural resources, increasing of reserves buffer zones and fencing of reserves.