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Analysis of two-dimensional dissociation constant of laterally mobile cell adhesion molecules.

We formulate a general analysis to determine the two-dimensional dissociation constant (2D Kd), and use this method to study the interaction of CD2-expressing T cells with glass-supported planar bilayers containing fluorescently labeled CD58, a CD2 counter-receptor. Both CD2 and CD58 are laterally mobile in their respective membranes. Adhesion is indicated by accumulation of CD2 and CD58 in the cell-bilayer contact area; adhesion molecule density and contact area size attain equilibrium within 40 min. The standard (Scatchard) analysis of solution-phase binding is not applicable to the case of laterally mobile adhesion molecules due to the dynamic nature of the interaction. We derive a new binding equation, B/F=[(Ntxf)/(KdxScell)]-[(Bxp)/Kd], where B and F are bound and free CD58 density in the contact area, respectively; Nt is CD2 molecule number per cell; f is CD2 fractional mobility; Scell is cell surface area; and p is the ratio of contact area at equilibrium to Scell. We use this analysis to determine that the 2D Kd for CD2-CD58 is 5.4-7.6 molecules/microm2. 2D Kd analysis provides a general and quantitative measure of the mechanisms regulating cell-cell adhesion.

CD2 Antigens↗

Comparison of a new dental trauma splint device (TTS) with three commonly used splinting techniques.

Splinting is the standard of care for stabilization of replanted or repositioned permanent teeth following trauma. The present experimental study compared four dental trauma splints in 10 volunteers. The evaluated splints included a wire-composite splint (WCS), a button-bracket splint (BS), a resin splint (RS), and a new device (TTS=Titanium Trauma Splint) specifically developed for splinting traumatized teeth. All splints were bonded to the labial surfaces of the maxillary lateral and central incisors. Splints were left in place for 1 week. After splint removal, the next splint was placed after a 1-week rest period. The sequence of splint application was randomized for each individual. The following parameters were assessed: tooth mobility with horizontal and vertical Periotest values (PTV) before and after splint application and splint removal, respectively; probing depths, plaque and bleeding on probing indices before splint application and removal, and chair time needed for splint application and removal. After splint application, horizontal PTV were significantly lower in central incisors for BS compared to TTS (P=0.04), and for RS compared to TTS (P=0.005) and to WCS (P=0.006). Reduction of lateral tooth mobility (=splint effect) expressed by the difference between horizontal pre- and postoperative PTV was significantly greater in RS compared to TTS and WCS (P<0.05) for central as well as for lateral incisors. However, changes of vertical tooth mobility were not significant across the splinting techniques. Periodontal parameters remained unchanged, reflecting the excellent oral hygiene by the study subjects. The chair time needed for splint application was significantly shorter for TTS (P<0.01). In conclusion, all tested splints appeared to maintain physiologic vertical and horizontal tooth mobility. However, the latter was critically reduced in RS splints.

Acrylic Resins↗

A quantitative structure property relationship study of electrophoretic mobility of analytes in capillary zone electrophoresis.

A quantitative structure property relationship (QSPR) is proposed to calculate the electrophoretic mobility of analytes in capillary zone electrophoresis. The proposed model employs logarithm of the electrophoretic mobility (ln micro) as dependent variable and partial charge (PQ), surface area (V(2/3)), total energy (TE), heat of formation (DeltaH(f)) and molecular refractivity (MR) as independent variables whose calculated using AM1 (Austin model 1) semi-empirical quantum mechanics method by HyperChem 7.0 software. The general form of the model is: ln micro =K(0)+K(1)PQ+K(2)V(2/3)+K(3)TE+K(4)DeltaH(f)+K(5)MR, where K(0)-K(5) are the model constants computed using a least-square method. The applicability of the model on real mobility data has been studied employing five experimental data sets of beta-blockers, benzoate derivatives, non-steroidal anti-inflammatory drugs, sulfonamides and amines in different buffers. The accuracy of the model is assessed using absolute average relative deviation (AARD) and the overall AARD value. The obtained AARD for the sets studied are 1.0 (N=10), 2.1 (N=26), 0.8 (N=11), 0.6 (N=13) and 2.7% (N=18), respectively, and the overall AARD is 1.4%. The model is cross-validated using one leave out technique and the obtained overall AARD is 1.8%. To further investigate on the applicability of the proposed model, the prediction capability of the model is evaluated by employing a minimum number of six experimental data points as training set, and predicting the mobility of other data points using trained models. The obtained overall AARD (for 48 predicted data points) is 5.6%.

Journal Article↗

[Mobilization and transplantation of autologous hematopoietic stem and progenitor cells from peripheral blood: principles and applications].

The use of peripheral blood progenitor cells instead of bone marrow to rescue hematopoiesis has dramatically reduced the morbidity and costs of high-dose chemo/radiotherapy. The time of aplasia after the myeloablative regimen is now only 8-14 days, reducing hospital stay and use of antibiotics and red cell or platelet transfusions. Sufficient progenitor and stem cells can be obtained with a small number of leukaphereses after mobilization in the blood by chemotherapy and hematopoietic growth factors or by growth factors alone. This method constitutes the treatment of choice for patients with relapse of Hodgkin's disease and of high-grade non-Hodgkin's lymphoma. Future developments will include the definition of new indications and the search for ways to eliminate tumor cells from the graft.

Antineoplastic Agents↗

Inhibition of Ca2+ mobilization by caffeine in a cultured vascular smooth muscle cell line (A7r5).

The effects of caffeine on the resting level and agonist-induced changes in intracellular calcium ([Ca2+]i) have been studied in the vascular smooth muscle cell line A7r5. Caffeine (1-30 mM) lowers the resting [Ca2+]i by reducing the entry of Ca2+ and inhibits completely the mobilization of Ca2+ by arginine vasopressin. Application of forskolin, to elevate cAMP, does not affect the resting level of Ca2+i but does abolish the agonist-induced rise. These data add to the complexity of caffeine-induced changes in [Ca2+]i and point to a possible interaction between cAMP and other second messenger systems mobilizing Ca2+i in this cell type.

Animals↗

The actions of caffeine and 2,3-butanedione monoxime on calcium transients in human vascular smooth muscle.

Ca2+ mobilization by membrane depolarization or histamine application, was measured in isolated human uterine artery smooth muscle cells. Nifedipine, a Ca2+ channel blocker, was found to inhibit the depolarization-induced increase in [Ca2+]i but not the histamine-induced increase. This suggests the presence of functional voltage-dependent Ca2+ channels and agonist-induced mobilization of Ca2+ via a different mechanism. Caffeine inhibited both the depolarization- and histamine-induced increases in intracellular calcium. The mechanisms of inhibition do not involve cAMP and point to a complex action of caffeine at multiple sites. The dephosphorylating agent 2,3-butanedione monoxime (BDM) was found to block voltage-dependent Ca2+ changes but not agonist-induced changes suggesting a role for phosphorylation in the regulation of the Ca2+ channels in these cells.

Caffeine↗

Mobility of Zn, Cd and Pb in soils as affected by poultry litter extract--I. Leaching in soil columns.

Application of poultry litter to cropland may increase metal mobility, because the soluble organic ligands in poultry litter can form water-soluble complexes with metals. In this study, one uncontaminated soil and two metal-contaminated soils were sampled. A portion of the uncontaminated soil was amended with Zn, Pb, and Cd at rates of 400, 200, and 8 mg kg(-1), respectively. Packed soil columns were leached with H2O, EDTA, CaCl2, and poultry litter extract (PLE) solutions separately. No leaching of Zn, Cd, and Pb with the PLE was found in the uncontaminated soil. The retention of PLE-borne Zn indicated the potential for Zn accumulation in the soil. A large portion of the metals was leached from the metal-amended soil, and EDTA solubilized more Zn, Cd, and Pb than CaCl2 and PLE. In the metal-contaminated soils, the leaching of Zn and Cd with PLE was consistently larger than that for CaCl2, indicating that these metals were mobilized by organic ligands. The PLE did not mobilize Pb in these soils. The utilization of poultry litter in metal-contaminated soils might accelerate the movement of Zn and Cd in soil profiles.

Journal Article↗

Ionic liquids as electrolytes for nonaqueous capillary electrophoresis.

Acetonitrile is a well-suited medium for nonaqueous capillary electroseparations and enables extending the range of applications of capillary electrophoresis (CE) techniques to more hydrophobic species. In this study, the dialkylimidazolium-based low temperature melting organic salts know as "ionic liquids" (ILs) are used as electrolytes. At room temperature these liquids are miscible with acetonitrile which makes it easy to use them for adjustment of analyte mobility and separation. The anionic part as well as the concentration of an IL influence the general electrophoretic mobility of the buffer system. The separation of different analytes is achieved because they become charged in the presence of ILs in separation media. There is also a possibility for a complex formation between the solute and the electrolyte which alters the mobility of the solute. A selected application of separations of phenols and aromatic acids will be discussed.

Electrolytes↗

Process analysis using ion mobility spectrometry.

Ion mobility spectrometry, originally used to detect chemical warfare agents, explosives and illegal drugs, is now frequently applied in the field of process analytics. The method combines both high sensitivity (detection limits down to the ng to pg per liter and ppb(v)/ppt(v) ranges) and relatively low technical expenditure with a high-speed data acquisition. In this paper, the working principles of IMS are summarized with respect to the advantages and disadvantages of the technique. Different ionization techniques, sample introduction methods and preseparation methods are considered. Proven applications of different types of ion mobility spectrometer (IMS) used at ISAS will be discussed in detail: monitoring of gas insulated substations, contamination in water, odoration of natural gas, human breath composition and metabolites of bacteria. The example applications discussed relate to purity (gas insulated substations), ecology (contamination of water resources), plants and person safety (odoration of natural gas), food quality control (molds and bacteria) and human health (breath analysis).

Bacteria↗

Assessment of arsenic mobility in the soils of some golf courses in South Florida.

High concentrations of arsenic have been detected in soils and underlying groundwater of some South Florida golf courses, indicating the possible impact of the application of arsenic-containing herbicides. The mobility of arsenic in the soils from selected golf courses was studied using a simple two-step sequential extraction procedure. Sodium nitrate (0.1 M), potassium dihydrogen phosphate (0.1 M) and concentrated nitric acid were used to obtain mobile, mobilizable, and pseudo total arsenic fractions. Soils were separated into fine (<0.25 mm) and large (0.25-0.75 mm) particle size fractions. Arsenic contents were correlated with the distribution of iron (R2=0.4827), manganese (R2=0.7674) and aluminum (R2=5459) in the particle size fractions, while such correlation was not observed for soil organic matter, indicating that the oxides/hydroxides of iron, manganese and aluminum control the distribution of arsenic in these soils. Sodium nitrate and potassium dihydrogen phosphate extractants used in this study extracted large portions of arsenic from most soil samples studied. This is especially true for the fine fraction where the extractable arsenic ranged from 9.2 to 51.3% with an average of 28.7 +/- 13.3%, whereas in the large fraction, arsenic ranged from 7.2 to 24.7% with an average of 15.4 +/- 6.4%. These extractants, however, release only small amounts of iron, manganese, and aluminum. It seems likely that arsenic can be released by sodium nitrate and potassium dihydrogen phosphate without significant dissolution of the oxides/hydroxides of iron, manganese, and aluminum in these soil samples.

Aluminum↗

Lymphedema, lipedema, and the open wound: the role of compression therapy.

As the science of wound healing has evolved over the past two decades, so has awareness of the "hidden epidemic" of lymphedema. Substantial information has been accumulated regarding the pathophysiology and therapy of lymphedema. Until recently, the relationship between wound healing and the negative effects of associated peri-wound lymphedema has received little attention. Identifying wound-related lymph stasis and safe mobilization of the fluid are fundamentals that must be addressed for proper therapy. Experience gained from the successful treatment of primary and secondary lymphedema has proven very useful in the applications to wound-related lymphedema. The mobilization of lymph fluid from the peri-wound area with the use of reasoned compression is essential for proper therapy of the open wound, as are appropriate bandage selection and safeguards for bandage application.

Adipocytes↗

Patterns of [Ca2+](i) mobilization and cell response in human spermatozoa exposed to progesterone.

Human spermatozoa stimulated with progesterone (a product of the cumulus and thus encountered by sperm prior to fertilization in vivo) apparently mobilize Ca(2+) and respond very differently according to the way in which the steroid is presented. A progesterone concentration ramp (0-3 microM) induces [Ca(2+)](i) oscillations (repetitive store mobilization) which modify flagellar beating, whereas bolus application of micromolar progesterone causes a single large transient (causing acrosome reaction) which is apparently dependent upon Ca(2+) influx. We have investigated Ca(2+)-mobilization and functional responses in human sperm exposed to 3 muM progesterone. The [Ca(2+)](i) response to progesterone was abolished by 4 min incubation in 0 Ca(2+) medium (2 mM EGTA) but in nominally Ca(2+)-free medium (no added Ca(2+); 0 EGTA) a smaller, slow response occurred. Single cell imaging showed a similar effect of nominally Ca(2+)-free medium and approximately 5% of cells generated a small transient even in the presence of EGTA. When cells were exposed to EGTA-containing saline (5 min) and then returned to nominally Ca(2+)-free medium before stimulation, the [Ca(2+)](i) transient was greatly delayed (approximately 50 s) and rise time was doubled in comparison to cells not subjected to EGTA pre-treatment. We conclude that mobilization of stored Ca(2+) contributes a 'slow' component to the progesterone-induced [Ca(2+)](i) transient and that incubation in EGTA-buffered saline is able rapidly to deplete this store. Analysis of flagellar activity induced by 3 muM progesterone showed an effect (modified beating) associated with the [Ca(2+)](i) transient, in >80% of cells bathed in nominally Ca(2+)-free medium. This was reduced greatly in cells subjected to 5 min EGTA pre-treatment. The store-mediated transient showed a pharmacological sensitivity similar to that of progesterone-induced [Ca(2+)](i) oscillations (consistent with filling of the store by an SPCA) suggesting that the transient induced by micromolar progesterone is a 'single shot' activation of the same store that generates Ca(2+) oscillations.

Acrosome↗

Extensive mobilization of the urethral plate and urethra for repair of hypospadias: the modified Barcat technique.

The modified Barcat technique was used to treat 168 patients with hypospadias during the last 5 years. The technique, which is applicable for distal hypospadias, uses apposing fully mobilized meatal based skin flaps. Cosmetic and functional results are excellent, with only 3.5% of the patients requiring reoperation. Because the surgical technique does not depend on the configuration of the glans, depth of the urethral groove, caliber of the urethral meatus or even whether the patient has been circumcised, it is particularly suitable for training residents who can perform a single operation to correct a wide variety of cosmetic deformities.

Child↗

Capabilities and potentialities of transverse pore gradient gel electrophoresis.

Transverse pore gradient gel electrophoresis is important as a tool for obtaining nonlinear Ferguson plots [log(mobility) vs. gel concentration], e.g. in application to DNA in polyacrylamide gels or to agarose gels, with the purpose of evaluating molecular properties (size, conformation, malleability) and gel fiber properties (fiber radius and length per unit volume). To date, it is capable of (i) yielding gel patterns ("Ferguson curves") of migration distance vs. predicted % T-range of the pore gradient, assuming its linearity; (ii) yielding information regarding molecular conformation from the intersection of Ferguson curves of unknowns (e.g. bent DNA) with those of standards; (iii) acquisition of Ferguson curves by computer, using prototype instrumentation; (iv) mathematical manipulation of acquired Ferguson curves to yielding Ferguson plots, providing that mobility in free solution has been assessed by capillary zone electrophoresis. The potentialities of the method remain unfulfilled to date due to (i) the unavailability, with a single exception, of an accurate and precise way to produce pore gradients of known shape; (ii) unavailability of a routinely applicable analysis for % T; (iii) unavailability of optimized, user-friendly and foolproof instrumentation for computer acquisition of Ferguson curves, including the present inapplicability of a commercially available electrophoresis apparatus with intermittent optical detection to transverse pore gradient gels; and (iv) unresolved problems in the statistical evaluation of Ferguson curves.

DNA↗

Molecular dynamics in living cells observed by fluorescence correlation spectroscopy with one- and two-photon excitation.

Multiphoton excitation (MPE) of fluorescent probes has become an attractive alternative in biological applications of laser scanning microscopy because many problems encountered in spectroscopic measurements of living tissue such as light scattering, autofluorescence, and photodamage can be reduced. The present study investigates the characteristics of two-photon excitation (2PE) in comparison with confocal one-photon excitation (1PE) for intracellular applications of fluorescence correlation spectroscopy (FCS). FCS is an attractive method of measuring molecular concentrations, mobility parameters, chemical kinetics, and fluorescence photophysics. Several FCS applications in mammalian and plant cells are outlined, to illustrate the capabilities of both 1PE and 2PE. Photophysical properties of fluorophores required for quantitative FCS in tissues are analyzed. Measurements in live cells and on cell membranes are feasible with reasonable signal-to-noise ratios, even with fluorophore concentrations as low as the single-molecule level in the sampling volume. Molecular mobilities can be measured over a wide range of characteristic time constants from approximately 10(-3) to 10(3) ms. While both excitation alternatives work well for intracellular FCS in thin preparations, 2PE can substantially improve signal quality in turbid preparations like plant cells and deep cell layers in tissue. At comparable signal levels, 2PE minimizes photobleaching in spatially restrictive cellular compartments, thereby preserving long-term signal acquisition.

Animals↗

Cardiac magnetic resonance in outpatients in Germany--indications, complications and protocol suggestions from a high-volume center.

BACKGROUND: Cardiac magnetic resonance (CMR) has developed into a routine examination in many centers in cardiology. However, there is little knowledge about its applicability in outpatients as a diagnostic tool for cardiovascular diseases. We report about the experiences in a high-volume cardiac imaging center and in a "mobile setting" in Germany and provide routinely used examination protocols. METHODS: 8976 patients referred for CMR from cardiologists, internal medicine practices and from general practitioners and 2200 patients examined in a "mobile" system by outpatient cardiologists were included in the study. Indications were as follows: 7672 (69%) examinations for myocardial ischemia and viability, 1313 (12%) for cardiac and pericardial inflammatory disease and cardiac mass, 976 (9%) for detection and quantification of heart valve disease and 466 (4%) for congenital heart disease. 697 (6%) were referred for other indication. Two independent readers performed image analysis of the 8976 patients in our center. RESULTS: Image quality was rated "excellent" in 90.6%, "good" in 8%, "fair" in 1.2% and "poor" in 0.2%. 0.0002% of all examinations were not assessable due to low image quality. Minor complications (temporarily, asymptomatic AV-blockade; mild chest pain and/or dyspnea; nausea) could be observed in 12% and resolved within few minutes. One patient experienced a grand mal seizure due to hyperventilation. 0.9% examinations had to be terminated untimely due to claustrophobia. CONCLUSION: CMR in outpatients is a widely used imaging modality in cardiology in Germany. A large variety of clinical questions may be answered by CMR with excellent image quality and without major complication. With user-adapted protocols, a rapid diagnosis is achieved even in outpatients in a "mobile" setting. Hence, CMR will increase its applicability as a routine imaging tool.

Clinical Protocols↗

Design and validation of a methodology using the International Classification of Diseases, 9th Revision, to identify secondary conditions in people with disabilities.

OBJECTIVE: To design and validate a methodology that identifies secondary conditions using International Classification of Disease, 9th Revision (ICD-9) codes. DESIGN: Secondary conditions were identified through a literature search and a survey of Washington State physiatrists. These conditions were translated into ICD-9 codes and this list was then validated against a national sample of Medicare survey respondents with differing levels of mobility and activities of daily living (ADL) disability. SETTING: National survey. PARTICIPANTS: Participants (N=9731) in the 1999 Medicare Current Beneficiary Survey with no, mild, moderate, and severe mobility and ADL disability. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURE: Percentage of survey respondents with a secondary condition. The secondary conditions were grouped into 4 categories: medical, psychosocial, musculoskeletal, and dysphagia related (problems associated with difficulty in swallowing). RESULTS: Our literature search and survey of 26 physiatrists identified 64 secondary conditions, including depression, decubitus ulcers, and deconditioning. Overall, 70.4% of all survey respondents were treated for a secondary condition. We found a significant relation between increasing mobility as well as ADL disability and increasing numbers of secondary conditions (chi 2 test for trend, P <.001). This relation existed for all categories of secondary conditions: medical (chi 2 test for trend, P <.001), psychosocial (chi 2 test for trend, P <.001), musculoskeletal (chi 2 test for trend, P <.001), and dysphagia related (chi 2 test for trend, P <.001). CONCLUSIONS: We created a valid ICD-9-based methodology that identified secondary conditions in Medicare survey respondents and discriminated between people with different degrees of disability. This methodology will be useful for health services researchers who study the frequency and impact of secondary conditions.

Disability Evaluation↗

On the electrophoretic mobility of biological cells.

A theoretical study has been performed on the electrophoretic mobility of a large colloidal particle with a surface charge layer as a model for biological cells. An approximate mobility expression is derived which is applicable to the general case of an arbitrary distribution of membrane-fixed charges. This expression consists of two terms: the first is a weighted average of potentials over the surface charge layer, while the second is that of the volume density of membrane-fixed charges and does not depend on the electrolyte concentration. At high electrolyte concentrations (corresponding to physiological conditions), where the potentials are very low owing to the shielding effects of electrolytes, the first term diminishes so that the mobility is determined mainly by the second term. This means that a particle with zero surface potential can exhibit a non-zero mobility, in contrast to the prediction from the classical Smoluchowski theory. Comparison is also made with the theory of Hermans and Fujita for the electrophoresis of polyelectrolytes.

Cell Membrane↗