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PubMed2004

Roles of coherent ongoing oscillations among dynamic cell assemblies in object perception.

I constructed a cortical neural network model and investigated possible roles of coherent ongoing oscillations in membrane potentials of neurons in object perception. The model has a hierarchical structure consisting of two lower networks and one higher network that are reciprocally connected via divergent/convergent projections. Information about features and their relationships (or objects) is encoded by the population activities of neurons (or dynamic cell assemblies) of the lower networks and the higher network, respectively. The ongoing state of the network is expressed by 'random itinerancy' among these dynamic cell assemblies. Under the ongoing state, the dynamic cell assemblies belonging to the same object are transiently linked across the networks and coherently oscillate at lower frequencies (approximately 15 Hz). When the model perceives a presented object, the dynamic cell assemblies corresponding to the object are persistently linked together across the networks and coherently oscillate at higher frequencies (approximately 40 Hz). When the feedback pathways are impaired, the dynamic phase transition from the slow- to fast-oscillations is not induced by the object presentation, keeping the lower frequency oscillations (approximately 15 Hz) where the activated dynamic cell assemblies oscillate incoherently. Reaction times to the object presentation are greatly reduced if the ongoing oscillation frequencies fall within a specific range (approximately 20-30 Hz). I suggest that coherent ongoing slow-oscillations in cortical activity may serve as a ready state for sensory input, whereby the brain can respond effectively to sensory stimulation. Top-down processing via feedback pathways may give an essential contribution to the induction of coherent fast-oscillations across multiple cortical areas, by which relevant features are effectively integrated into a unified percept when stimulated with a sensory object.

Action Potentials↗
PubMed2005

Nonlinear dynamical analysis of noisy time series.

Empirical time series in the life sciences are often nonstationary and have small signal-to-noise ratios, making it difficult to accurately detect and characterize dynamical structure. The usual response to high noise is averaging, but time domain averaging is inappropriate, especially when the dynamics are nonlinear. We review alternative delay-space averaging methods based on the topology and short-term predictability of nonlinear dynamics and illustrate their application using the TISEAN software (Hegger, Kantz & Schreiber, 1999). The methods were applied to a Lorenz series, which resembles the dynamics found by Kelly, Heathcote, Heath and Longstaff (2001) in series of decision times. The Lorenz series was corrupted with up to 80% additive Gaussian noise, a lower signal-to-noise ratio than has been used in any previous test of these methods, but consistent with Kelly et al.'s data. Prediction methods performed the best for detecting nonstationarity and nonlinear dynamics, and optimal predictability provided an objective criterion for setting the parameters required by the analyses. Local linear filtering methods performed best for characterization, producing informative plots that revealed the nature of the underlying dynamics. These results suggest that a methodology based on delay-space averaging and prediction could be useful with noisy empirical data series.

Animals↗
PubMed2006

Adjusting behavioral methods when applying nonlinear dynamical measures to stimulus rate.

A nonlinear dynamical perspective addresses an area of behavior that tends to be ignored: trial-to-trial variability under seemingly identical conditions. Data exhibiting sequence effects are excellent candidates for a nonlinear dynamical analysis, because of their dependence on previous trial events. When applying nonlinear dynamical tools to behavioral data, the assumptions underlying the tools may require changes in experimental methodology. In this paper, the assumptions of trial timing and their potential impact on nonlinear dynamical analysis are explored. This is done with the intent of initiating a discussion on the appropriate control of trial timing. To examine these issues, each participant's response times on simple two-choice tasks were treated as a time series and submitted along with two comparisons to two nonlinear dynamical measures. Two experimental variables, stimulus rate and stimulus-response mapping, were manipulated in order to examine the issue of controlling trial timing via stimulus rate. Significant differences between the observed and comparison time series were found for estimated dimensionalities(m). Differences in dimensionality(m) estimates were also found between the experimental variables. Of the two methods for controlling stimulus rate, response-stimulus-interval and inter-stimulus-interval, the latter is recommended when applying nonlinear dynamical measures.

Analysis of Variance↗
PubMed1996

In vivo dynamics of myosin II in Dictyostelium by fluorescent analogue cytochemistry.

We used fluorescent analogue cytochemistry to study in vivo dynamics of myosin II in Dictyostelium discoideum. We labeled myosin with biotin or tetramethyl-rhodamine iodoacetamide (IATR). The labeled myosin shows normal activities as reversible filament assembly and Ca2+ and actin-activatable Mg(2+)-ATPase. We used the biotin-myosin as a probe examining the effects of microinjection on the amoebae and the ability to associate with endogenous actin cytoskeleton. The biotin-myosin incorporates into certain actin populations and localizes to the cortex with the highest accumulation in the posterior end of polarized amoebae. The dynamics in live amoebae were probed by TR-myosin. We monitored the dynamics for a long period to determined the dynamic reorganization corresponding specific cellular behaviors. The TR-myosin converges into a discrete actin- and myosin-rich structure located at the posterior end ("myosin-organizing center"). The rod-shaped TR-myosin exhibits linear orderly arrays emanating from the organizing center which extend about two-thirds of the cell length. The myosin arrays show a dynamic reorganization when the amoebae move. To examine if the observed myosin dynamics are related to filamentous (F-) actin, we disrupted the F-actin by cytochalasin D. The ratioed image of TR-myosin (vs. FITC-dextran) demonstrates that myosin in these cells accumulates in the cortex but does not form the organizing center. Overall, the results suggest that the filamentous myosin organizes into orderly arrays in the live cytoplasm and its translocation occurs by means of F-actin cables, converging into the organizing center.

Animals↗
PubMed1998

Relationship between microtubule dynamics and lamellipodium formation revealed by direct imaging of microtubules in cells treated with nocodazole or taxol.

Microtubules (MTs) contribute to the directional locomotion of many cell types through an unknown mechanism. Previously, we showed that low concentrations (<200 nM) of nocodazole or taxol reduced the rate of locomotion of NRK fibroblasts over 60% without altering MT polymer level [Liao et al., 1995: J. Cell Sci. 108:3473-3483]. In this paper, we directly measured the dynamics of MTs in migrating NRK cells injected with rhodamine tubulin and treated with low concentrations of nocodazole or taxol. Both drug treatments caused statistically significant reductions (approx. twofold) in growth and shortening rates and less dramatic effects on rescue and catastrophe transition frequencies. The percent time MTs were inactive (i.e., paused) increased greater than twofold in nocodazole- and taxol-treated cells, while the percent time growing was substantially reduced. Three parameters of MT dynamics were linearly related to the rates of locomotion determined previously: rate of shortening, percent time pausing and percent time growing. The number of MTs that came within 1 microm of the leading edge was reduced in drug-treated cells, suggesting that reduced MT dynamics may affect actin arrays necessary for cell locomotion. We examined two such structures, lamellipodium and adhesion plaques, and found that lamellipodia area was coordinately reduced with MT dynamics. No effect was detected on adhesion plaque density or distribution. In time-lapse recordings, MTs did not penetrate into the lamellipodium of untreated cells, suggesting that MTs affect lamellipodia either through their interaction with factors at the base of the lamellipodium or by releasing factors that diffuse into the lamellipodia. In support of the latter hypothesis, when all MTs were rapidly depolymerized by 20 microM nocodazole, we detected the rapid formation of exaggerated protrusions from the leading edge of the cell. Our results show for the first time a linear relationship between MT dynamics and the formation of the lamellipodium and support the idea that MT dynamics may contribute to cell locomotion by regulating the size of the lamellipodium, perhaps through diffusable factors.

Animals↗
PubMed1996

Dynamic synapse: a new concept of neural representation and computation.

Presynaptic mechanisms influencing the probability of neurotransmitter release from an axon terminal, such as facilitation, augmentation, and presynaptic feedback inhibition, are fundamental features of biological neurons and are cardinal physiological properties of synaptic connections in the hippocampus. The consequence of these presynaptic mechanisms is that the probability of release becomes a function of the temporal pattern of action potential occurrence, and hence, the strength of a given synapse varies upon the arrival of each action potential invading the terminal region. From the perspective of neural information processing, the capability of dynamically tuning the synaptic strength as a function of the level of neuronal activation gives rise to a significant representational and processing power of temporal spike patterns at the synaptic level. Furthermore, there is an exponential growth in such computational power when the specific dynamics of presynaptic mechanisms varies quantitatively across axon terminals of a single neuron, a recently established characteristic of hippocampal synapses. During learning, alterations in the presynaptic mechanisms lead to different pattern transformation functions, whereas changes in the postsynaptic mechanisms determine how the synaptic signals are to be combined. We demonstrate the computational capability of dynamic synapses by performing speech recognition from unprocessed, noisy raw waveforms of words spoken by multiple speakers with a simple neural network consisting of a small number of neurons connected with synapses incorporating dynamically determined probability of release. The dynamics included in the model are consistent with available experimental data on hippocampal neurons in that parameter values were chosen so as to be consistent with time constants of facilitative and inhibitory processes governing the dynamics of hippocampal synaptic transmission studied using nonlinear systems analytic procedures.

Acoustic Stimulation↗
PubMed1998

Cell membrane lipid molecular dynamics in a solenoid versus a magnetically shielded room.

The generalized polarization function of the fluorescent probe 2-dimethylamino-6-lauroylnaphthalene has been used to evaluate the lipid dynamics in Friend erythroleukemia cell membrane. The values of this function varied during the culture growth cycle, showing decreased lipid dynamics 24-48 h from the cell seeding. When the cycle occurred in a solenoid producing a magnetic field of 70 microT at 50 Hz in addition to the 45 microT DC of the earth (short-term 4-day exposure), the membrane lipid dynamics during this same time-period decreased by about 10% (P < .04). After long-term (184 days) or extremely long-term (395 days) exposure of the cells to the magnetic field, little additional variation in the membrane lipid dynamics was observed, suggesting an adaptation phenomenon. A variation of membrane lipid dynamics was also observed due to in vitro cell differentiation (P < .02). Nevertheless, the exposure of both undifferentiating and differentiating cells to a highly attenuated magnetic field in a magnetically shielded room (20 nT DC plus 2.5 pT AC) did not induce any modification of membrane lipid dynamics.

Animals↗
PubMed2000

Dynamic exercise therapy for rheumatoid arthritis.

OBJECTIVES: To assess the effects of dynamic exercise therapy in improving joint mobility, muscle strength, aerobic capacity and daily functioning in patients with rheumatoid arthritis (RA). In addition, possible unwanted effects such as an increase in pain, disease activity and radiological progression were studied. SEARCH STRATEGY: We searched the Cochrane Musculoskeletal trials register, the Cochrane Controlled trials Register, Medline, Embase and SCISEARCH databases up to may 1997 in order to controlled trials on the effect of exercise therapy. SELECTION CRITERIA: Randomized trials on the effect of dynamic exercise therapy in RA patients with an exercise program fulfilling the following criteria: a) intensity level such that heart rates exceeded 60% of maximal heart rate during at least 20 minutes, b) exercise frequency of two session per week, and c) duration of intervention of greater than six weeks DATA COLLECTION AND ANALYSIS: Two blinded reviewers independently selected eligible studies, rated the methodological quality and extracted data. MAIN RESULTS: Six out of 30 identified controlled trials met the inclusion criteria. Four of the six included studies fulfilled at least seven out of 10 methodological criteria. Due to heterogeneity in outcome measures, data could not be pooled. The results suggest that dynamic exercise therapy is effective at increasing aerobic capacity and muscle strength. No detrimental effects on disease activity and pain were observed. The effects of dynamic exercise therapy on functional ability and radiological progression are unclear. REVIEWER'S CONCLUSIONS: Dynamic exercise therapy has a positive effect on physical capacity. Research on the long-term effect of dynamic exercise therapy on radiological progression and functional ability is needed.

Arthritis, Rheumatoid↗
PubMedDate not supplied

Morphological variation and airflow dynamics in the human nose.

Airflow dynamics are recognized as being important to the functioning of the human nose in conditioning and filtering inspired air, yet these dynamics are poorly understood. Despite considerable research on airflow dynamics by otolaryngologists, respiratory physiologists, and toxicologists, major disagreements remain about the nature of airflow in the human nose. Specifically, there is little consensus about the character of nasal airflow regimes (laminar or turbulent) and about the major pathways of airflow through the internal chamber. Additionally, a number of features in the human nose have been argued to enhance airflow turbulence, thus increasing the exposure of moving air to the nasal mucosa and facilitating heat and moisture exchange in cold and/or dry climates. These features include: an inferior orientation of the nares; a nasal sill that is high relative to the floor of the internal nasal chamber; a nasal valve that is small in cross-sectional area relative to that of the internal chamber; and large, projecting conchae. The claim that these features affect airflow dynamics has never been tested. To clarify the nature of human nasal airflow and to test these claims of functional significance to nasal variation, we studied airflow across physiological flow rates using water and dye flowing through anatomically accurate acrylic models of human nasal air passageways (with adjustment of water flow rates to maintain dynamic similarity). The models were derived from direct casting of the nasal passageways of 10 Caucasian ("leptorrhine") cadavers (six male, four female). Measures of naris angle, nasal sill height, nasal valve area relative to internal chamber cross-sectional area, and relative projection of the inferior and middle turbinates were taken directly on the resulting casts. The relationships between aspects of nasal morphology and turbulent air flow were evaluated by examining the flow regimes (laminar, semiturbulent, or turbulent) at varying flow rates, with the expectation that the greater the development of the proposed turbulence-enhancing features the slower the flow rate at which flow would shift from one regime to another. Flow characteristics (both flow regimes and principal pathways) were highly variable within our sample. The relative projection of the inferior turbinate was the only variable that significantly affected the flow rate at which flow became turbulent. However, more projecting turbinates appear to laminate flow rather than to induce turbulence. Nostril orientation was moderately correlated with flow dynamics (with more inferiorly directed nares producing turbulence at slower flow rates), but this correlation was not statistically significant. Relative nasal valve area and nasal sill height were unrelated to turbulence in our models.

Algorithms↗
PubMed2002

Global and local motions in ribonuclease A: a molecular dynamics study.

The understanding of protein dynamics is one of the major goals of structural biology. A direct link between protein dynamics and function has been provided by x-ray studies performed on ribonuclease A (RNase A) (B. F. Rasmussen et al., Nature, 1992, Vol. 357, pp. 423-424; L. Vitagliano et al., Proteins: Structure, Function, and Genetics, 2002, Vol. 46, pp. 97-104). Here we report a 3 ns molecular dynamics simulation of RNase A in water aimed at characterizing the dynamical behavior of the enzyme. The analysis of local and global motions provides interesting insight on the dynamics/function relationship of RNase A. In agreement with previous crystallographic reports, the present study confirms that the RNase A active site is constituted by rigid (His12, Asn44, Thr45) and flexible (Lys41, Asp83, His119, Asp121) residues. The analysis of the global motions, performed using essential dynamics, shows that the two beta-sheet regions of RNase A move coherently in opposite directions, thus modifying solvent accessibility of the active site, and that the mixed alpha/3(10)-helix (residues 50-60) behaves as a mechanical hinge during the breathing motion of the protein. These data demonstrate that this motion, essential for RNase A substrate binding and release, is an intrinsic dynamical property of the ligand-free enzyme.

Animals↗
PubMed1991

Virtual rigid body dynamics.

An important direction in biological simulations is the development of methods that permit the study of larger systems and/or longer simulation time scales than is currently feasible by molecular dynamics. One such method designed with this objective in mind is stochastic boundary molecular dynamics (SBMD). SBMD was developed for the characterization of spatially localized processes in proteins, and has been shown to successfully reproduce structural and dynamical properties of these macromolecules, as compared to a molecular dynamics control simulation, when concerted or global motions are not present. The virtual rigid body dynamics method presented in this work extends the range of applicability of the SBMD method, by providing a framework to include these important long time scale conformational transitions. In this paper we describe the two-step implementation of the virtual rigid body model: first, the reduction of the full atomic representation to a reduced particle (virtual bond) model, and second, the propagation of the dynamics of flexibly connected rigid bodies containing virtual atom sites.

Biopolymers↗
PubMed2004

Use of the time-signal intensity curve from dynamic magnetic resonance imaging to evaluate remnant pancreatic fibrosis after pancreaticojejunostomy in patients undergoing pancreaticoduodenectomy.

BACKGROUND: This study assessed the value of the time-signal intensity curve (TIC) obtained from dynamic magnetic resonance imaging (MRI) in the evaluation of remnant pancreatic fibrosis after pancreaticojejunostomy in patients undergoing pancreaticoduodenectomy. METHODS: Two modes of pancreaticojejunostomy-duct-to-mucosa anastomosis (DMA; 24 patients) and pancreatojejunoserosal anastomosis (PJSA; 22 patients)-were used in 46 consecutive patients undergoing pancreaticoduodenectomy. All patients underwent dynamic contrast-enhanced MRI of the pancreas before pancreaticoduodenectomy. Retrospective review of the pancreatic magnetic resonance images and histological examination of the pancreas were performed, and the patterns of TICs from dynamic MRI were compared with the degree of pancreatic fibrosis. Dynamic MRI of the residual pancreas was carried out for 1-3 years after pancreaticoduodenectomy in 26 patients (14 DMA, 12 PJSA) who had a histologically verified normal pancreas with no fibrosis at the time of pancreaticoduodenectomy. RESULTS: Evaluation of preoperative dynamic magnetic resonance images showed that a pancreatic TIC with a rapid rise to a peak followed by a rapid decline (type I) was characteristic of a normal pancreas without fibrosis. Pancreatic TICs with a slow rise to a peak followed by a slow decline or a plateau (types II and III) indicated a fibrotic pancreas. Postoperative pancreatic MRI demonstrated that six of 12 patients with a PJSA had a type II TIC, whereas 12 of 14 patients with a DMA had a type I curve (P = 0.046). CONCLUSION: The TIC obtained from dynamic MRI is a reliable indicator of fibrosis in the remnant pancreas after pancreaticoduodenectomy. Use of a DMA was associated with a lower risk of pancreatic fibrosis 1-3 years after surgery than a PJSA.

Adult↗
PubMed2006

Regulation of microtubule dynamics in 3T3 fibroblasts by Rho family GTPases.

To get insight into the action of Rho GTPases on the microtubule system we investigated the effects of Cdc42, Rac1, and RhoA on the dynamics of microtubules in Swiss 3T3 fibroblasts. In control cells microtubule ends were dynamic: plus ends frequently switched between growth, shortening and pauses; the growth phase predominated over shortening. Free minus ends of microtubules depolymerized rapidly and never grew. Free microtubules were short-lived, and the microtubule network was organized into a radial array. In serum-starved cells microtubule ends became more stable: although plus ends still transited between growth and shortening, polymerization and depolymerization excursions became shorter and balanced each other. Microtubule minus ends were also stabilized. Consequently lifespan of free microtubules increased and microtubule array changed its radial pattern into a random one. Activation of Cdc42 and Rac1 in serum-starved cells promoted dynamic behavior of microtubule plus and minus ends, while inhibition of these GTPases in serum-grown cells suppressed microtubule dynamics and mimicked all effects of serum starvation. Activation of RhoA in serum-grown cells had effects similar to Cdc42 /Rac1 inactivation: it suppressed the dynamics of plus and minus ends, reduced the length of growth and shrinking episodes, and disrupted the radial organization of microtubules. However, in contrast to Cdc42 and Rac1 inactivation, active RhoA had no effect on the balance between microtubule growth and shortening. We conclude that Cdc42 and Rac1 have similar stimulating effects on microtubule dynamics while RhoA acts in an opposite way.

Animals↗
PubMed1988

Dynamic instability and motile events of native microtubules from squid axoplasm.

Native microtubules from extruded axoplasm of squid giant axons were used as a paradigm to characterize the motion of organelles along free microtubules and to study the dynamics of microtubule length changes. The motion of large round organelles was visualized by AVEC-DIC microscopy and analyzed at a temporal resolution of 10 frames per second. The movements were smooth and showed no major changes in velocity or direction. During translocation, the organelles paused very rarely. Superimposed on the rather constant mean velocity was a velocity fluctuation, which indicated that the organelles are subject to considerable thermal motion during translocation. Evidence for a regular low-frequency oscillation was not found. The thermal motion was anisotropic such that axial motion was less restricted than lateral motion. We conclude that the crossbridge connecting the moving organelle to the microtubule has a flexible region that behaves like a hinge, which permits preferential movement in the direction parallel to the microtubule. The dynamic changes in length of native microtubules were studied at a temporal resolution of 1 Hz. About 98% of the native microtubules maintained their length ("stable" microtubules), while 2% showed phases of growing and/or shrinking typical for dynamic instability ("dynamic" microtubules). Gliding and organelle motion were not influenced by dynamic length changes. Transitions between growing and shrinking phases were low-frequency events (1-10 minutes per cycle). However, a new type of microtubule length fluctuation, which occurred at a high frequency (a few seconds per cycle), was detected. The length changes were in the 1-3 micron range. The latter events were very prominent at the (+) ends. It appears that the native axonal microtubules are much more stable than the purified microtubules and the microtubules of cultured cells that have been studied thus far. Potential mechanisms accounting for the three states of microtubule stability are discussed. These studies show that the native microtubules from squid giant axons are a very useful paradigm for studying microtubule-related motility events and microtubule dynamics.

Animals↗
PubMed1993

Unambiguous classification of microtubule-ends in vitro: dynamic properties of the plus- and minus-ends.

To understand the mechanism of dynamic instability of microtubule growth and shortening, one needs a means of reliably determining the polarity of the microtubules under investigation. Sea urchin sperm-tail axonemal fragments nucleate the growth of both plus-ended and minus-ended microtubules, but their polarity is not apparent by video-enhanced DIC microscopy. The polarity of a microtubule is usually assessed by observing differences between the rates and lengths of growth and shortening excursions of the two ends. In practice, though, a significant fraction of the population of microtubules displays characteristics intermediate between the average characteristics of either end, thereby escaping classification. Excluding these "intermediate" microtubules from the measured populations introduces bias into the understanding of microtubule dynamic instability. We circumvent this problem by making use of the plus-end directed movement of the microtubule-dependent molecular motor kinesin to determine the polarity of any given microtubule unambiguously. Carboxylated-microspheres coated with kinesin, which are clearly visible by DIC microscopy, were used to determine the polarity of a microtubule. The dynamics were then observed. Kinesin was found to have no marked effect on dynamic instability. By this technique, we show that the distributions of properties that describe microtubule dynamic instability (rates and lengths of growth and shortening as well as frequencies of interconversion between these phases) of plus-ends overlap to a significant extent with those of minus-ends. It is this overlap that obscures the usual classification of the ends. Therefore, models describing microtubule dynamic instability need to incorporate the broad and overlapping range of properties of the two ends.

Animals↗
PubMed1995

beta-Tubulin mutation suppresses microtubule dynamics in vitro and slows mitosis in vivo.

Microtubule (MT) dynamics vary both spatially and temporally within cells and are thought to be important for proper MT cellular function. Because MT dynamics appear to be closely tied to the guanosine triphosphatase (GTPase) activity of beta-tubulin subunits, we examined the importance of MT dynamics in the budding yeast S. cerevisiae by introducing a T107K point mutation into a region of the single beta-tubulin gene, TUB2, known to affect the assembly-dependent GTPase activity of MTs in vitro. Analysis of MT dynamic behavior by video-enhanced differential interference contrast microscopy, revealed that T107K subunits slowed both the growth rates and catastrophic disassembly rates of individual MTs in vitro. In haploid cells tub2-T107K is lethal; but in tub2-T107K/tub2-590 heterozygotes the mutation is viable, dominant, and slows cell-cycle progression through mitosis, without causing wholesale disruption of cellular MTs. The correlation between the slower growing and shortening rates of MTs in vitro, and the slower mitosis in vivo suggests that MT dynamics are important in budding yeast and may regulate the rate of nuclear movement and segregation. The slower mitosis in mutant cells did not result in premature cytokinesis and cell death, further suggesting that cell-cycle control mechanisms "sense" the mitotic slowdown, possibly by monitoring MT dynamics directly.

Base Sequence↗
PubMed1996

An analysis of the dynamic range and linearity of an infrared DNA sequencer.

We are currently developing genotyping software and protocols for use on the Li-Cor model 4000(L) infrared fluorescence DNA sequencers. During the development of the genotyping software it became apparent that the potential dynamic range of the instrument was not being realized when the data collection parameters were optimized to produce a high-contrast image on the computer screen. In particular, we were unable to obtain peaks with signal to noise (S/N) greater than about 30:1 and the stronger peaks often saturated the detector. Because the numerical dynamic range available in 16-bit data collection mode exceeds 65000, our limited dynamic range of about 30 in actual data was somewhat puzzling. Hence, we undertook a study to explore the dynamic range and linearity of the Li-Cor DNA sequencer in order to minimize background fluorescence and noise as well as maximize the available S/N. Data is presented on the background and noise using different polyacrylamide gel mixes with various signal gain and offset values, and the relative contribution of the glass, gel and instrumentation to the background fluorescence is discussed. Based on these results, optimum gain and offset values were determined that maintain the background fluorescence at approximately 1-2% of the maximum dynamic range with a minimum amount of noise. Using these optimized values, we determined the detection system is linear over three orders of magnitude by titrating known quantities of an infrared fluorophore-labeled primer. In addition, we were able to detect approximately 15.2 amoles of labeled primer. The results provided by this study establish a set of guidelines for evaluating how to set the signal gain and offset in order to minimize the background and maximize the S/N and dynamic range of the Li-Cor sequencer.

DNA↗
PubMed2004

Capillary electrophoresis analysis of a wide variety of seized drugs using the same capillary with dynamic coatings.

Capillary electrophoresis methodology is presented for the routine analysis of a wide variety of seized drugs using the same capillary with dynamic coatings and multiple run buffers. The types of exhibits analyzed using diode array UV detection include phenethylamines, cocaine, oxycodone, heroin, lysergic acid diethylamide (LSD), opium, hallucinogenic mushrooms, and gamma-hydroxybutyrate-gamma-butyrolactone (GHB-GBL). Both qualitative and quantitative analyses are achieved using run buffers that contain additives that provide for secondary equilibrium and/or dynamic coating of the capillary. Dynamic coating of the capillary surface is accomplished by rapid flushes of 0.1 N sodium hydroxide, water, buffer containing polycation coating reagent, and a buffer containing a polyanionic coating reagent (with or without cyclodextrin(s)) or a micelle coating reagent. Dynamic coating with a polyanionic coating reagent is used for the analysis of moderately basic seized drugs and adulterants. The use of cyclodextrin in the run buffer not only allows for chiral analysis but also greatly enhances separation selectivity for achiral solutes. A capillary dynamically coated with a micelle allows for the analysis of neutral, acidic, and weakly basic drugs (GHB, GBL and neutral, acidic, and weakly basic adulterants). Dynamic coating, which gives rise to a relatively high and robust electroosmotic flow at pH < 7, allows for rapid, precise and reproducible separations. For a wide variety of drugs, excellent linearity and migration time precision and good peak area precision (external and internal standard) is obtained. Quantitative results for synthetic mixtures are in good agreement with actual values. Screening for adulterants is greatly enhanced by the use of automated library searches.

Alkaloids↗