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PubMed2005

Transendocardial and transepicardial intramyocardial fibroblast growth factor-2 administration: myocardial and tissue distribution.

Effective local delivery to the heart remains an obstacle to successful therapeutic application of a number of drugs and biological agents. This study was designed to study and optimize the delivery characteristics of transendocardial intramyocardial (IM) administration, determine myocardial deposition and retention over time, and compare it to transepicardial IM injection. Thirty-nine pigs were used for the study (15 for catheter optimization, 15 for transendocardial IM delivery, and 9 for transepicardial IM delivery). (125)I-Fibroblast growth factor-2 (FGF2) (25 microCi) was used as the prototype molecule. Tissue and myocardial distribution was determined at 1 and 24 h and 7 days. Using 1-h (125)I-FGF2 myocardial deposition as a parameter for delivery efficiency, the optimal needle length and delivery volume for transendocardial based delivery were determined to be 6 mm and 0.1 ml, respectively. Using these parameters for endocardial delivery, (125)I-FGF2 cardiac activity was 18.01 +/- 3.84% of delivered activity at 1 h, 11.65 +/- 5.17% at 24 h, and 2.32 +/- 0.87% at 7 days in ischemic animals. Studies in nonischemic animals produced similar results. For transepicardial delivery, (125)I-FGF2 cardiac-specific activity was 23.14 +/- 12.67% for the 6-mm needle, declining to 12.32 +/- 8.50% at 24 h, and did not significantly differ from values obtained following transendocardial delivery. Thus, optimized transendocardial intramyocardial delivery using Biosense guidance results in efficient delivery of FGF2 to the target myocardium that is comparable with transepicardial delivery, both providing markedly higher myocardial deposition and retention and lower systemic recirculation of FGF2 than intracoronary, intrapericardial, or intravenous delivery. However, myocardial distribution is limited to injection sites.

Animals↗
PubMed2026

Landscape of retron diversity across the SPIRE microbial metagenome resource reveals candidate novel type XI-like lineages.

Retrons are bacterial genetic elements encoding a specialized reverse transcriptase (RT) that synthesizes multicopy single-stranded DNA and are increasingly recognized as components of bacterial anti-phage defense systems. However, their diversity and ecological distribution across large-scale genomic resources remain poorly characterized. Here, we surveyed retron RTs across the SPIRE representative metagenome collection, a non-redundant, species-level data set spanning diverse microbial habitats. Using a curated panel of type-specific hidden Markov models, we identified retrons representing all canonical types together with additional divergent lineages. Retron distribution showed strong taxonomic and ecological structuring, with some groups restricted to specific bacterial phyla, whereas others were broadly distributed across environmental categories. Systematic novelty assessment identified two candidate type XI-like lineages, TXI_C2like and TXI_noncan_h, characterized by protease-independent architectures and distinct accessory modules associated with WYL- and DnaB_C-containing proteins, respectively. De novo covariance-based analyses further identified candidate msr/msd-like non-coding RNA structures in both lineages, supporting conservation of the canonical RT-ncRNA organizational framework despite extensive sequence divergence. Together, these findings expand the known diversity of retron systems and identify type XI-like retrons as a dynamic and previously underexplored evolutionary group.IMPORTANCERetrons are bacterial genetic elements that are increasingly exploited as programmable tools for genome editing, molecular recording, and biosensing in addition to their natural role in anti-phage defense. Despite this growing biotechnological interest, the true diversity of retrons across the bacterial world has remained largely unmapped. By mining a resource of over 100,000 processed microbial metagenomes, we uncovered thousands of retron sequences spanning known types as well as previously unrecognized lineages and found that their distribution is strongly shaped by both bacterial taxonomy and ecological niche. Among these, we identified two candidate new lineages related to type XI retrons that lack the protease domain typical of this group but instead carry distinct accessory proteins, expanding the known architectural diversity of these systems. These findings broaden the catalog of retron diversity available for functional characterization and biotechnological engineering and provide a framework for prioritizing candidate lineages for future experimental validation.

effectors↗
PubMed2004

Left septal atrial flutter: electrophysiology, anatomy, and results of ablation.

BACKGROUND: We describe the clinical and electrophysiological characteristics of a novel macroreentrant form of left atrial flutter circuit. METHODS AND RESULTS: A total of 11 patients were included in the study. The mean tachycardia cycle length was 278+/-41 ms. Nine of the 11 patients were treated with antiarrhythmic drugs at the time of the study for concomitant atrial fibrillation. With the use of entrainment pacing and either the CARTO Biosense mapping system (9 patients) or conventional mapping (2 patients), the flutter circuit was found to rotate around the left septum primum with a critical isthmus located between the pulmonary veins posteriorly and/or mitral annulus anteriorly and the septum primum. In 5 patients, radiofrequency ablation was performed from the septum primum to the right inferior pulmonary vein (group 1), and in 6 patients, a lesion was made from the septum primum to the mitral annulus (group 2). After a follow-up of 13+/-6 months, 2 patients in group 1 and all patients in group 2 remained in sinus rhythm without recurrence. CONCLUSIONS: Slowing of electric conduction in the left atrial septum due to antiarrhythmic drugs and/or atrial myopathy seems to promote left septal atrial flutter. Radiofrequency ablation of this arrhythmia is usually effective and safe. A line of block between the septum primum and the mitral annulus proved to be effective for cure of tachycardia.

Aged↗
PubMed1999

Electroanatomic left ventricular mapping in the porcine model of healed anterior myocardial infarction. Correlation with intracardiac echocardiography and pathological analysis.

BACKGROUND: Catheter ablation for ventricular tachycardia in healed infarction is limited to patients with inducible, tolerated arrhythmias. Strategies that would allow mapping during sinus rhythm might obviate this limitation. METHODS AND RESULTS: Two sets of experiments were performed in adult pigs to refine a new technique for left ventricular mapping. First, detailed endocardial maps were done in 5 normal pigs and 7 pigs 6 to 10 weeks after left anterior descending coronary artery infarction to characterize electrograms in normal and infarcted tissue by electroanatomic mapping (CARTO, Biosense). Electrogram recording sites were verified by intracardiac echo (ICE, 9 MHz) and grouped by location: infarct (area of akinesis by ICE), border (0.5-cm perimeter of akinetic area), and remote. Compared with remote sites, electrograms from infarct sites had smaller amplitudes (1.2+/-0.5 versus 5.1+/-2.1 mV, P<0.001), longer durations (74.2+/-26.3 versus 36.3+/-6.4 ms, P<0.001), and more frequent notched or late components. Border zone electrograms were intermediate in amplitude and duration. Second, infarct characterization by electroanatomic mapping was compared with pathological (exclusion of triphenyltetrazolium chloride staining) and ICE measurements. Infarct size by pathology correlated with the area defined by contiguous electrograms with amplitude </=1 mV (r=0.98, P=0.0001). Infarct size by ICE imaging correlated with the area defined by contiguous electrograms with amplitude </=2 mV (r=0.95, P=0.0016). CONCLUSIONS: Electroanatomic mapping during sinus rhythm allows accurate 3D characterization of infarct architecture and defines the relationship of electrophysiological and anatomic abnormalities. This technique may prove useful in devising anatomically based strategies for ablation of ventricular tachycardia.

Animals↗
PubMed1997

Three-dimensional mapping of the common atrial flutter circuit in the right atrium.

BACKGROUND: The full circuit of common atrial flutter using conventional methods of sequential or multielectrode activation mapping is not completely understood. METHODS AND RESULTS: We performed three-dimensional right atrial endocardial activation mapping during common counterclockwise atrial flutter in 17 patients (16 men, 1 woman; mean age, 53+/-11 years) by using the Cordis-Biosense EP Navigation system and assessed the distribution of estimated conduction velocities and double and fractionated potentials. ECG flutter wave morphologies were compared with activation patterns. Points (91+/-29) were sequentially acquired covering 88+/-11% of the flutter cycle length of 239+/-22 ms. A wide and variable posterior zone of double and fractionated potentials coincided with blocking and colliding wave fronts and formed the posterior limit of the circuit. A progressively widening septal (sep) wave front ascending from just beyond the coronary sinus ostium, passed cranially as a broad front anterior to the superior vena cava (SVC) in 14 patients, whereas fusion around the SVC formed the superior (sup) limb of the circuit in 3. Bounded anteriorly by the tricuspid valve, the wave front descended down the lateral (lat) aspect of the right atrium before completing the circuit in all cases through the inferior vena cava-tricuspid annulus isthmus. The estimated conduction velocity in the medial isthmus (0.6+/-0.3 m/s) was lower than in the other limbs of the circuit (sup=1+/-0.5 m/s, lat=1+/-0.5 m/s, sep=0.9+/-0.4 m/s, P=.05). Double and fractionated potentials were constant and more prevalent in the posterior right atrium. ECG flutter wave morphology did not correlate with three-dimensional activation maps. CONCLUSIONS: Interindividual variations occur in the right atrial circuit of common atrial flutter, with constant activation through the cavotricuspid isthmus. A variable zone of block forms the posterior limit. Fusion around the SVC can occur, and ascending medial septal activation does not follow a consistent pattern.

Adult↗
PubMed1999

High-density mapping of activation through an incomplete isthmus ablation line.

BACKGROUND: Activation mechanisms through gaps in ablation lines and resulting electrograms are poorly understood. METHODS AND RESULTS: Eight patients (all men; age, 59+/-9 years) were studied during a recurrence of typical atrial flutter (cycle length, 233+/-19 ms) after a previous catheter ablation in the cavotricuspid isthmus. High-density 3-dimensional mapping of the isthmus was performed with the Cordis-Biosense EP Navigation system, and local conduction velocity (CV) was estimated. Maps created with 96+/-19 points revealed 0.8+/-0.3-cm gaps of recovered conduction in the ablation line. A broad wave front entered the lateral isthmus with a CV of 1.8+/-0.7 m/s, halted on the lesion line, and penetrated slowly through the gap with a CV of 0.3+/-0.1 m/s. Activation then curved and returned antidromically to activate the downstream flank of the line with a CV of 1.1+/-0.7 m/s. This front fused downstream of the line with slow transverse activation (CV, 0.4+/-0.3 m/s) parallel to it. The ablation line was demarcated by an incomplete line of convergent double potentials with isoelectric intervals (from 123+/-34 to 62+/-16 ms); each potential corresponded to local activation upstream and downstream of the lesions, while the intervening delay was produced by slow conduction through the gap combined with the progressively longer curved pathway of downstream antidromic activation as a function of distance from the gap. CONCLUSIONS: High-density isthmus mapping during recurrent flutter indicates slow conduction through gaps of recovered conduction of varying dimensions in the ablation line followed by a curved front of activation antidromically activating its downstream flank, this detour producing wide double potentials on the line.

Atrial Flutter↗
PubMed2005

Successful radiofrequency ablation in patients with previous atrial fibrillation results in a significant decrease in left atrial size.

BACKGROUND: The objective of the present study was to evaluate the relation between freedom from atrial fibrillation (AF) and left atrial (LA) size in patients who underwent circumferential pulmonary vein (PV) isolation and LA ablation. METHODS AND RESULTS: One hundred five consecutive patients with symptomatic and drug-refractory paroxysmal or persistent AF were included in the present study. The mean age was 52+/-9.5 years (range, 27 to 75 years); 74 patients (70%) were male. Paroxysmal AF was present in 52 (49.5%) and persistent AF in 53 (50.5%) patients. Mean AF duration was 6.0+/-5.1 years in the paroxysmal AF group and 7.6+/-6.0 years in the persistent AF group. A 3D electroanatomic map of the LA including the PV ostia was constructed with a nonfluoroscopic navigation system (Carto, Biosense Webster). Left- and right-sided PVs were encircled by continuous radiofrequency ablation lines. We performed 128 ablation procedures in 105 patients, ie, 23 redo procedures. The mean long-term follow-up duration was 14.6+/-4.9 months (range, 6 to 24 months). Sinus rhythm was present in 45 patients (86.5%) in the paroxysmal AF group and in 41 patients (77.3%) in the persistent AF group at the latest follow-up. Six months after ablation, LA dimension in the persistent AF subjects who remained in sinus rhythm decreased from 44.0+/-5.8 to 40+/-4.5 mm (range, 31 to 51 mm). In contrast, in patients with recurrences of AF, LA dimension increased from 45+/-6.5 to 49+/-5.4 mm (range, 32 to 59 mm). In the successfully treated paroxysmal AF group, LA dimension decreased from 40.5+/-4.4 to 37.5+/-3.5 mm (P<0.01). CONCLUSIONS: In radiofrequency ablation of AF using an electroanatomic approach, there is a statistically significant relationship between medium-term procedural success and LA size: persistent sinus rhythm is associated with reduced and recurrent AF with increased LA dimensions.

Adult↗
PubMed2006

Carbon nanotube-templated assembly of protein.

This paper describes a novel general strategy for fabricating protein-polyion multilayers by electrostatic layer-by-layer (LBL) self-assembly on carbon nanotube templates. Such a noncovalent functionalization method is important for preserving the activity of biomolecular, the mechanical, and electrical properties of carbon nanotubes. Glucose oxidase and poly(diallyldimethylammonium) chloride polymer (PDDA) were used as models to investigate the LBL process on CNT templates. High-resolution TEM and electrochemical characterization confirm the formation of LBL nanostructures on carboxyl functionalized carbon nanotubes. We have also demonstrated the applications of these nanoshell bioreactors to direct electrochemistry of protein and biosensing. This strategy can be applied to assemble other biological molecules such as antibody, antigen, and DNA for wide bioassay applications.

Coated Materials, Biocompatible↗
PubMed2006

Nanopore Detector based analysis of single-molecule conformational kinetics and binding interactions.

BACKGROUND: A Nanopore Detector provides a means to transduce single molecule events into observable channel current changes. Nanopore-based detection can report directly, or indirectly, on single molecule kinetics. The nanopore-based detector can directly measure molecular characteristics in terms of the blockade properties of individual molecules--this is possible due to the kinetic information that is embedded in the blockade measurements, where the adsorption-desorption history of the molecule to the surrounding channel, and the configurational changes in the molecule itself, imprint on the ionic flow through the channel. This rich source of information offers prospects for DNA sequencing and single nucleotide polymorphism (SNP) analysis. A nanopore-based detector can also measure molecular characteristics indirectly, by using a reporter molecule that binds to certain molecules, with subsequent distinctive blockade by the bound-molecule complex. RESULTS: It is hypothesized that reaction histories of individual molecules can be observed on model DNA/DNA, DNA/Protein, and Protein/Protein systems. Preliminary results are all consistent with this hypothesis. Nanopore detection capabilities are also described for highly discriminatory biosensing, binding strength characterization, and rapid immunological screening. CONCLUSION: In essence, the heart of chemistry is now accessible to a new, single-molecule, observation method that can track both external molecular binding states, and internal conformation states.

Base Sequence↗
PubMed2006

Surge capacity for health care systems: early detection, methodologies, and process.

Excessive demand on hospital services from large-scale emergencies is something that every emergency department health care provider and hospital administrator knows could happen at any time. Nowhere in this country have we recently faced a disaster of the magnitude of concern we now face involving agents of mass destruction or social disruption, especially those in the area of infectious diseases and radiological materials. The war on terrorism is not a conventional war, and terrorists may use any means of convenience to carry out their objectives in an unpredictable time line. Have we adequately prepared for the potentially excessive surge in demand for medical services that a large-scale event could bring to our medical care system? Are our emergency departments ready for such events? Surveillance systems, such as BioWatch, BioSense, the National Biosurveillance Integration System, and the countermeasure program BioShield, offer hope that we will be able to meet these new challenges.

Delivery of Health Care↗
PubMed2004

Two-dimensional phase unwrapping with a multichannel least-mean-square algorithm.

This paper presents a computationally efficient two-dimensional phase-unwrapping method based on a multichannel least-mean-square algorithm. The performance of the proposed method is evaluated by applying phase unwrapping to several simulated very noisy images and to a genuine noisy interferometrical image taken from a five-step phase-shift interferogram obtained from a surface plasmon resonance imaging biosensing experiment. The results confirm that the proposed method is more widely applicable, more computationally efficient, and more robust in the presence of noise than the representative methods presented in this paper.

Journal Article↗
PubMed2006

Surface plasmon resonance sensors based on uniform-waist tapered fibers in a reflective configuration.

We present a configuration for surface plasmon resonance sensors based on uniform-waist tapered optical fibers and reflective elements. Once the fiber is tapered fulfilling the adiabatic criterion, a multilayer including a metallic medium is asymmetrically deposited on the uniform waist of the fiber. This feature provides the resonant excitation of multiple surface plasma waves. In addition, a mirror is produced at the fiber tip by a chemical Tollens reaction. In this way, the sensor operates in a reflective mode, more convenient for dip probes. When these sensors are spectrally interrogated, a high sensitivity of 10-4 refractive index units per nanometer is attained. These devices can be advantageously used for any kind of chemical sensing and biosensing.

Journal Article↗
PubMed2003

Light scattering by microscopic spheres behind a glass-air interface.

Scattering of light from single spheres placed behind a glass-air interface with light incident through the glass is examined. This scattering is investigated for both p- and s-polarized light incident at angles below the glass-air critical angle. The intensity of light scattered into the air half-space from each sphere is measured as a function of scattering angle, and this response is compared in situ with the background scatter produced by the planar substrate. A detailed comparison between data and established theory are thereby obtained. This system is of interest in the field of optical biosensing.

Journal Article↗
PubMed2005

Investigation of dual-channel fiber-optic surface plasmon resonance sensing for biological applications.

A dual-channel fiber-optic sensor based on surface plasmon resonance (SPR) for self-referencing refractive-index measurements has been proposed. Most applications of fiber-optic SPR sensors are designed to measure the refractive index of a liquid or gas sample by measuring the signal from a single surface, the sensitivity and stability of which is easily affected by the fluctuation of external environmental conditions. We have designed a dual-channel fiber-optic surface sensor with two independent SPR signals from two areas of the same probe. A prototype sensor was fabricated and characterized. The preliminary experimental results demonstrate the characteristic responses of both SPR signals from two channels that independently correspond to the refractive index changes in the liquid samples with which they are in contact. The design could be extended to a multichannel sensor with further developments. The experimental results confirmed that one channel can be used as a reference sensor that could compensate for unexpected changes in bulk refraction or temperature and develop this sensor as a practicable high-sensitivity biosensing device.

Journal Article↗
PubMed1999

Immunocytochemical distribution of cytochrome P4501A (CYP1A) in developing gilthead seabream, Sparus aurata.

CYP1A is a major inducible enzyme in the metabolism of xenobiotic substrates. In this paper we investigate by means of immunohistochemistry, the tissue distribution of constitutive cytochrome P4501A (CYP1A) during the period of endogenous nutrition (from hatching until day 4) in developing gilthead seabream, Sparus aurata larvae. For this purpose, a polyclonal antiserum (BN-1, Biosense Laboratories) directed against conserved piscine CYP1A sequences was used on paraffin-embedded sections from seabream larvae. From hatching onward, CYP1A immunoreactivity was observed in the following tissues and cells: syncytial, oil-globule envelopes and matrix of the yolk-sac, kidney (epithelia of renal tubules), cardiac muscle cells, skin epidermal cells, troncal musculature, enterocytes of different intestinal regions, goblet cells of the bucco-pharyngeal region, gill epithelial cells and the endothelia of the vascular system of various tissues (especially from liver and brain). Moreover, eye (retina), olfactory epithelium and some positive nerve fibers located in the proximity of the olfactory bulbs and running ventrally toward the posterior brain were strongly CYP1A immunoreactive. In general, the intensity of immunostaining increased with larval development.

Animals↗
PubMed2006

The role of flexible tethers in multiple ligand-receptor bond formation between curved surfaces.

Ligands mounted to surfaces via extensible tethers are present in nature and represent a growing class of molecules used to engineer adhesion in drug targeting, biosensing, self-assembling nanostructures, and in other biophysical research. Using a continuum approach with geometric and thermodynamic arguments, we derive a number of analytical expressions that relate key properties of single-tethered ligand-receptor interactions to multiple bond formation between curved surfaces. The theoretical predictions are in good agreement with measurements made with the surface forces apparatus. We establish that, when ligated, many tethers commonly used in biophysical research exhibit a discrete binding range that can be accurately measured with force spectroscopy. The distribution of bound ligated tethers is independent of the surfaces' interaction radius, R. The bridging force scales linearly with R, the tether's effective spring constant and grafting density, and with the ligand-receptor bond energy when the surfaces are in direct contact. These results are contrasted to bridging forces that evolve between plane-parallel geometries. Last, we show how our simple analytical reductions can be used to predict adhesive forces for STEALTH liposomes and other targeted and self-assembled nanoparticles.

Binding Sites↗
PubMed2006

Structural stabilization and functional improvement of horseradish peroxidase upon modification of accessible lysines: experiments and simulation.

Horseradish peroxidase (HRP) is an important heme enzyme with enormous medical diagnostic, biosensing, and biotechnological applications. Thus, any improvement in the applicability and stability of the enzyme is potentially interesting. We previously reported that covalent attachment of an electron relay (anthraquinone 2-carboxylic acid) to the surface-exposed Lys residues successfully improves electron transfer properties of HRP. Here we investigated structural and functional consequences of this modification, which alters three accessible charged lysines (Lys-174, Lys-232, and Lys-241) to the hydrophobic anthraquinolysine residues. Thermal denaturation and thermoinactivation studies demonstrated that this kind of modification enhances the conformational and operational stability of HRP. The melting temperature increased 3 degrees C and the catalytic efficiency enhanced by 80%. Fluorescence and circular dichroism investigations suggest that the modified HRP benefits from enhanced aromatic packing and more buried hydrophobic patches as compared to the native one. Molecular dynamics simulations showed that modification improves the accessibility of His-42 and the heme prosthetic group to the peroxide and aromatic substrates, respectively. Additionally, the hydrophobic patch, which functions as a binding site or trap for reducing aromatic substrates, is more extended in the modified enzyme. In summary, this modification produces a new derivative of HRP with enhanced electron transfer properties, catalytic efficiency, and stability for biotechnological applications.

Binding Sites↗
PubMed2005

Edge plane pyrolytic graphite electrodes in electroanalysis: an overview.

The recent development, behavior and scope of edge plane pyrolytic graphite electrodes in electroanalysis are overviewed. Similarities to, and advantages, over multi-walled CNT modified electrodes are noted and the wide scope of applications, ranging through gas sensing, stripping voltammetry and biosensing, illustrated.

Ascorbic Acid↗