Cys mutants in functional regions of Sticholysin I clarify the participation of these residues in pore formation

Experimental evidence shows that the mechanism of pore formation by actinoporins is a multistep process, involving binding of the water-soluble monomer to the membrane and subsequent oligomerization on the membrane surface, leading to the formation of a functional pore. However, as for other eukaryotic pore-forming toxins, the molecular details of the mechanism of membrane insertion and oligomerization are not clear. In order to obtain further insight with regard to the structure-function relationship in sticholysins, we designed and produced three cysteine mutants of recombinant sticholysin I (rStI) in relevant functional regions for membrane interaction: StI E2C and StI F15C (in the N-terminal region) and StI R52C (in the membrane binding site). The conformational characterization derived from fluorescence and CD spectroscopic studies of StI E2C, StI F15C and StI R52C suggests that replacement of these residues by Cys in rStI did not noticeably change the conformation of the protein. The substitution by Cys of Arg(52) in the phosphocholine-binding site, provoked noticeable changes in rStI permeabilizing activity; however, the substitutions in the N-terminal region (Glu(2), Phe(15)) did not modify the toxin`s permeabilizing ability. The presence of a dimerized population stabilized by a disulfide bond in the StI E2C mutant showed higher pore-forming activity than when the protein is in the monomeric state, suggesting that sticholysins pre-ensembled at the N-terminal region could facilitate pore formation. (C) 2011 Elsevier Ltd. All rights reserved.

Effect of flavonoids on 2 `-deoxyguanosine and DNA oxidation caused by singlet molecular oxygen

Antioxidant potential is generally investigated by assaying the ability of a compound to protect biological systems from free radicals. However, non-radical reactive oxygen species can also be harmful. Singlet molecular oxygen ((1)O(2)) is generated by energy transfer to molecular oxygen. The resulting (1)O(2) is able to oxidize the nucleoside 2`-deoxyguanosine (dGuo), which leads to the formation of 8-oxo-7,8-dihydro-2`-deoxyguanosine (8-oxodGuo) and spiroiminodihydantoin 2`-deoxyribonucleoside diastereomers (dSp) in an aqueous solution. The main objective of the present study was to verify whether the presence of flavonoids (flavone, apigenin, quercetin, morin and catechin) at different concentrations could protect dGuo from (1)O(2) damage. Of the tested flavonoids, flavone possessed antioxidant activity, as determined by a decrease in the formation of both products. Apigenin, morin, quercetin and catechin all increased the formation of 8-oxodGuo at a concentration of 100 mu M. The quantification of plasmid strand breaks after treatment with formamidopyrimidine-DNA glycosylase showed that flavone protected and quercetin and catechin enhanced DNA oxidation. Our results show that compounds, such as flavonoids, may affect the product distribution of (1)O(2)-mediated oxidation of dGuo, and, in particular, high concentrations of flavonoids with hydroxyl groups in their structure lead to an increase in the formation of the mutagenic lesion 8-oxodGuo. (C) 2010 Elsevier Ltd. All rights reserved.

Single-wall carbon nanotubes modified with organic dyes: Synthesis, characterization and potential cytotoxic effects

This work deals with the covalent functionalization of single-wall carbon nanotubes (SWNTs) with phenosafranine (PS) and Nile Blue (NB) dyes. These dyes can act as photosensitizers in energy and electron transfer reactions, with a potential to be applied in photodynamic therapy. Several changes in the characteristic Raman vibrational features of the dyes suggest that a covalent modification of the nanotubes with the organic dyes occurs. Specifically, the vibrational modes assigned to the NH(2) moieties of the dyes are seen to disappear in the SWNT-dye nanocomposites, corroborating the bond formation between amine groups in the dyes and carboxyl groups in the oxidized nanotubes. The X-ray absorption (XANES) data also show, that the intense band at 398.6 eV attributed to 1s -> 2p pi* transition of the nitrogen of the aromatic PS ring, is shifted due to the bonding with the carbonic structure of the SWNTs. The cytotoxicity data of dyes-modified SWNT composites in the presence and absence of light shows that the SWNT-NB (4 mu g/mL) composite presents a good photodynamic effect, namely a low toxicity in the dark, higher toxicity in the presence of light and also a reduced dye photobleaching by auto-oxidation. (C) 2010 Elsevier B.V. All rights reserved.

Simple telemedicine for developing regions: Camera phones and paper-based microfluidic devices for real-time, off-site diagnosis

This article describes a prototype system for quantifying bioassays and for exchanging the results of the assays digitally with physicians located off-site. The system uses paper-based microfluidic devices for running multiple assays simultaneously, camera phones or portable scanners for digitizing the intensity of color associated with each colorimetric assay, and established communications infrastructure for transferring the digital information from the assay site to an off-site laboratory for analysis by a trained medical professional; the diagnosis then can be returned directly to the healthcare provider in the field. The microfluidic devices were fabricated in paper using photolithography and were functionalized with reagents for colorimetric assays. The results of the assays were quantified by comparing the intensities of the color developed in each assay with those of calibration curves. An example of this system quantified clinically relevant concentrations of glucose and protein in artificial urine. The combination of patterned paper, a portable method for obtaining digital images, and a method for exchanging results of the assays with off-site diagnosticians offers new opportunities for inexpensive monitoring of health, especially in situations that require physicians to travel to patients (e.g., in the developing world, in emergency management, and during field operations by the military) to obtain diagnostic information that might be obtained more effectively by less valuable personnel.

Diagnostics for the Developing World: Microfluidic Paper-Based Analytical Devices

Microfluidic paper-based analytical devices (mu PADs) are a new class of point-of-care diagnostic devices that are inexpensive, easy to use, and designed specifically for use in developing countries. (To listen to a podcast about this feature, please go to the Analytical Chemistry multimedia page at pubs.acs.org/page/ancham/audio/index.html.)

Paper-Based ELISA

Bill & Melinda Gates Foundation[51308]

Understanding Wax Printing: A Simple Micropatterning Process for Paper-Based Microfluidics

This technical note describes a detailed study on wax printing, a simple and inexpensive method for fabricating microfluidic devices in paper using a commercially available printer and hot plate. The printer prints patterns of solid wax on the surface of the paper, and the hot plate melts the wax so that it penetrates the full thickness of the paper. This process creates complete hydrophobic barriers in paper that define hydrophilic channels, fluid reservoirs, and reaction zones. The design of each device was based on a simple equation that accounts for the spreading of molten wax in paper.

Paper Microzone Plates

This paper describes 96- and 384-microzone plates fabricated in paper as alternatives to conventional multi-well plates fabricated in molded polymers. Paper-based plates are functionally related to plastic well plates, but they offer new capabilities. For example, paper-microzone plates are thin (similar to 180 mu m), require small volumes of sample (5 mu L per zone), and can be manufactured from inexpensive materials ($0.05 per plate). The paper-based plates are fabricated by patterning sheets of paper, using photolithography, into hydrophilic zones surrounded by hydrophobic polymeric barriers. This photolithography used an inexpensive formulation photoresist that allows rapid (similar to 15 min) prototyping of paper-based plates. These plates are compatible with conventional microplate readers for quantitative absorbance and fluorescence measurements. The limit of detection per zone loaded for fluorescence was 125 fmol for fluorescein isothiocyanate-labeled bovine serum albumin, and this level corresponds to 0.02 the quantity of analyte per well used to achieve comparable signal-to-noise in a 96-well plastic plate (using a solution of 25 nM labeled protein). The limits of detection for absorbance on paper was aproximately 50 pmol per zone for both Coomassie Brilliant Blue and Amaranth dyes; these values were 0.4 that required for the plastic plate. Demonstration of quantitative colorimetric correlations using a scanner or camera to image the zones and to measure the intensity of color, makes it possible to conduct assays without a microplate reader.

1989-1990 Yearbook

Vol. 8, Sept. 1990; 136 p.; b&w, color photographs TOC: Activities & Events…17 / Clubs & Programs…33 / Graduation…49 CREDITS: FACULTY ADVISOR, Vincent Banrey; PROJECT DIRECTOR, Catherine Whan; EDITORS, Luis Lopez; ASST. EDITOR, Rodolfo Medina; ART: Rodolfo Medina (Cover Design; Back Endsheet; Division Pages), Peter Abbate (Front Endsheet). GRAPHIC ARTIST: Rodolfo "Funky Cold' Medina. LAYOUT DESIGNERS: Vincent Banrey, Marino "Tito" Cabrera, Ann Marie Edwards, Charletin Enock, Ana Lisa Gonzalez, Edward Hollins, Luis Lopez, Peter Martinez, Rodolfo Medina, JilI Nage, Angela ?, Keith ?, Daly Rodriguez, Catherine Whan. WRITERS: Marino "Tito" Cabrera, Vincent Cousin, Edward Hollins, Luis Lopez, JilI Nage, Cathy Passiglia, Eartha Porter, Daly Rodriguez, Christiana Sommerville, Catherine Whan. PHOTOGRAPHERS: Peter Abbate, Marino "Tito" Cabrera, Charletin Enock, Milton Ferriera, Fran Gibson, Alexandra Gomez, Bernadette Henry, Edward Hollins, Juan Jiminez, Umoja Kwanguvu, Luis Lopez, Rodolfo Medina, Allen Scribner, Frank Tocco, Catherine Wran, Alan O. Young. Special Thanks to Cathy Soria of Classic Studio (as well as Frank & Tommy).

On Real-Time Monitoring And Control For Efficient Management Of Drinking Water Networks: Barcelona Case Study

Drinking water utilities in urban areas are focused on finding smart solutions facing new challenges in their real-time operation because of limited water resources, intensive energy requirements, a growing population, a costly and ageing infrastructure, increasingly stringent regulations, and increased attention towards the environmental impact of water use. Such challenges force water managers to monitor and control not only water supply and distribution, but also consumer demand. This paper presents and discusses novel methodologies and procedures towards an integrated water resource management system based on advanced ICT technologies of automation and telecommunications for largely improving the efficiency of drinking water networks (DWN) in terms of water use, energy consumption, water loss minimization, and water quality guarantees. In particular, the paper addresses the first results of the European project EFFINET (FP7-ICT2011-8-318556) devoted to the monitoring and control of the DWN in Barcelona (Spain). Results are split in two levels according to different management objectives: (i) the monitoring level is concerned with all the aspects involved in the observation of the current state of a system and the detection/diagnosis of abnormal situations. It is achieved through sensors and communications technology, together with mathematical models; (ii) the control level is concerned with computing the best suitable and admissible control strategies for network actuators as to optimize a given set of operational goals related to the performance of the overall system. This level covers the network control (optimal management of water and energy) and the demand management (smart metering, efficient supply). The consideration of the Barcelona DWN as the case study will allow to prove the general applicability of the proposed integrated ICT solutions and their effectiveness in the management of DWN, with considerable savings of electricity costs and reduced water loss while ensuring the high European standards of water quality to citizens.