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PubMed1989

Evidence for a sodium/calcium exchanger and voltage-dependent calcium channels in adipocytes.

The objective of these studies is to identify and characterize Ca2+-transport systems that may be of potential importance in the action of Ca2+-mobilizing hormones in the adipocyte. Using the Ca2+-sensitive photoprotein, aequorin, [Ca2+]i was estimated to be 0.15 microM, assuming an intracellular [Mg2+] of 1 mM. Substitution of Na+ with choline+ caused a transient increase in [Ca2+]i which was inversely related to extracellular [Na+], consistent with operation of a mediated Na+-Ca2+ exchange system. The stoichiometry was 3Na+:Ca2+. Elevation of extracellular K+ caused an increase in [Ca2+]i that was blocked by the Ca2+ channel antagonist, diltiazem, by omitting extracellular Ca2+, or by substituting Sr2+ for Ca2+. These findings indicate the presence of an Na+-Ca2+ exchanger and voltage-sensitive Ca2+ channels in adipocytes.

Adipose Tissue↗
PubMed2006

Hematopoietic stem cell mobilization after rat partial orthotopic liver transplantation.

BACKGROUND: On the basis of the recently recognized potential of hematopoietic stem cells (HSC) to give rise to hepatocytes, we investigated the possibility that HSC could be mobilized and home to the injured liver promoting tissue repair after 50% partial orthotopic liver transplantation (PLTx) in the rat. METHODS: Using sex-mismatched (female to male) syngeneic SD rats, we performed 50% PLTx or whole orthotopic liver transplantation (WLTx) versus 50% partial hepatectomy (PHx) and sham operation (O). Elements with stem cell markers were detected in peripheral blood (PB) and in the liver. Liver injury and regeneration were estimated. The sex-determining region for Y chromosome gene (SRY) was used to define cell origin by in situ hybridization in liver sections. RESULTS: Comparison of WLTx and PHx groups showed a lower survival rate (50%), in the PLTx group were (P<.05). Further, the liver injury was more serious and the levels of serum biochemical parameters were higher. Compared with PHx groups, on days 3 and 5 postoperatively, the mitosis index and the expression of PCNA were lower among the PLTx groups. Compared with WLTx and sham operation groups, beta2m-/Thy-1.1+, CD34+ cells in PB in PLTx groups and PHx were increased on day 1 postoperatively and decreased on the following days. Compared with PHx groups, beta2m-/Thy-1.1+, CD34+ cells were higher in PLTx. The CD34-, c-kit-, and Thy-1.1-positive cells detected in portal tract areas peaked during 3 to 5 days postoperatively in PLTx. Few SRY+ cells were detected in PLTx liver grafts. CONCLUSIONS: beta2m-/Thy-1.1+ and CD34+ stem cells mobilized after PLTx and PHx may be related to the reduced-size liver. Few HSC are involved in liver regeneration in PLTx.

Animals↗
PubMed1998

Measurement of facial soft tissue mobility in man.

OBJECTIVE: The assessment of facial mobility is a key element in the treatment of patients with facial motor deficits. In this study, we explored the utility of a three-dimensional tracking system in the measurement of facial movements. METHODS AND RESULTS: First, the three-dimensional movement of potentially stable facial soft-tissue, headcap, and dental landmarks was measured with respect to a fixed space frame. Based on the assumption that the dental landmarks are stable, their motion during a series of standardized facial animations was subtracted from that of the facial and headcap landmarks to estimate their movement within the face. This residual movement was used to determine which points are relatively stable (< or = 1.5 mm of movement) and which are not (> or = 1.5 mm of movement). Headcap landmarks were found to be suitable as references during smile, cheek puff, and lip purse animations, and during talking. In contrast, skin-based landmarks were unsuitable as references because of their considerable and highly variable movement during facial animation. Second, the facial movements of patients with obvious facial deformities were compared with those of matched controls to characterize the face validity of three-dimensional tracking. In all instances, pictures that appear to be characteristic of the various functional deficits emerged. CONCLUSIONS: Our results argue that tracking instrumentation is a potentially useful tool in the measurement of facial mobility.

Adolescent↗
PubMed2002

High-performance liquid chromatography with electrochemical detection applied to the analysis of 3,4-dihydroxymethamphetamine in human plasma and urine.

Metabolic activation in the disposition of 3,4-methylenedioxymethamphetamine (MDMA, "ecstasy") has been implicated in some of its pharmacological and toxicological effects, with the major metabolite 3,4-dihydroxymethamphetamine (HHMA) as a putative toxicant through the formation of thioether adducts. We describe the first validated method for HHMA determination based on acid hydrolysis of plasma and urine samples, further extraction by a solid-phase strong cation-exchange resin (SCX, benzenesulfonic acid), and analysis of extracts by high-performance liquid chromatography with electrochemical detection. The chromatographic separation was performed in an n-butyl-silane (C4) column and the mobile phase was a mixture of 0.1 M sodium acetate containing 0.1 M 1-octanesulphonic acid and 4 mM EDTA (pH 3.1) and acetonitrile (82:18, v/v). Compounds were monitored with an electrochemical cell (working potentials 1 and 2, +0.05 and +0.35 V, respectively, gain 60 microA). A mobile phase conditioning cell with a potential set at +0.40 V was connected between the pumping system and the injector. Calibration curves were linear within the working concentration ranges of 50-1000 microg/L for urine and plasma. Limits of detection and quantification were 10.5 and 31.8 microg/L for urine and 9.2 and 28.2 microg/L for plasma. Recoveries for HHMA and DHBA (3,4-dihydroxybenzylamine, internal standard) were close to 50% for both biological matrices. Intermediate precision and inter-day accuracy were within 3.9-6.5% and 7.4-15.3% for urine and 5.0-10.8% and 9.2-13.4% for plasma.

Calibration↗
PubMed1981

Mobile equivalent heart generator (migrating dipole).

The study is concerned with quantitative mapping of electrocardiographic potentials on the human body surface at various applications of equivalent multipole heart generators. The accuracy or reproduction of the electrical heart field with the application of mobile and immobile dipole and quadrupole generators is compared. The immobile equivalent generator (EG) is fixed in the geometrical centre of the trunk, whereas the mobile EG follows the path of the electrical heart centre during the heart cycle. Four healthy subjects and two patients with permanent postmyocardial ECG changes were examined. On transition from the immobile to the mobile model, in both groups of probands the relative contribution of the quadrupole component to the total potential of the body surface steeply diminished, with consequent substantial increase in accuracy of the dipole heart description. The dipole following the path of the electric heart centre during the heart cycle represents one of the most promising models, and deserves attentive clinical testing.

Electrocardiography↗
PubMed2006

Do mobile 'phones have a detrimental impact on auditory function?

Mobile telephones are now an integral part of modern telecommunications. The widespread use of mobile telephones has given rise to concern about the potential influences of electromagnetic fields (EMFs) on human health. The effects of EMFs on human beings are under investigation. Anatomically, the ear is in close proximity to the mobile telephone during use. The effect of mobile phones on auditory function has rarely been studied. The aim of this review is to determine if there is evidence that mobile phones have a detrimental effect on auditory function.

Animals↗
PubMed2002

Embryonic expression of the luteinizing hormone beta gene appears to be coupled to the transient appearance of p8, a high mobility group-related transcription factor.

A comparison between two pituitary-derived cell lines (alpha T3-1 and L beta T2) that represent gonadotropes at early and late stages of development, respectively, was performed to further elucidate the genomic repertoire required for gonadotrope specification and luteinizing hormone beta (LH beta) gene expression. One isolated clone that displayed higher expression levels in L beta T2 cells encodes p8, a high mobility group-like protein with mitogenic potential that is up-regulated in response to proapoptotic stimuli and in some developing tissues. To test the functional significance of this factor in developing gonadotropes, a knockdown of p8 in L beta T2 cells was generated. The loss of p8 mRNA correlated with loss of endogenous LH beta mRNA and the loss of activity of a transfected LH beta promoter-driven reporter, even upon treatment with gonadotropin-releasing hormone. In addition, expression of p8 mRNA in developing mouse pituitary glands mirrored its expression in the gonadotrope-derived cell lines and coincided with the first detectable appearance of LH beta mRNA. In contrast, p8 mRNA was undetectable in the pituitary glands of normal adults. Taken together, our data indicate that p8 is a stage-specific component of the gonadotrope transcriptome that may play a functional role in the initiation of LH beta gene expression during embryonic cellular differentiation.

Cell Line↗
PubMed1996

Electrophoretic mobility of biological cells in asymmetric electrolyte solutions.

The electrophoretic mobility of a particle covered by a membrane in an a:b electrolyte solution is modeled theoretically. The membrane, which simulates the surface of a biological cell, is ion-penetrable, and carries homogeneously distributed negative fixed charges. An approximate expression for the electrophoretic mobility is derived. Based on the results of numerical simulation, we conclude the following: (1) The absolute Donnan potential increases with the concentration of the fixed charges C0, but decreases with the ionic strength I. (2) The greater the valence of cation alpha, the lower the absolute potential distribution. (3) The greater the C0, the greater the absolute mobility of a particle, magnitude of mu, and the greater the friction coefficient of the membrane phase gamma, the smaller the magnitude of mu. (4) A large I or a large a leads to a small magnitude of mu. (5) The greater the ratio (permittivity of solution/permittivity of membrane phase), the smaller the magnitude of mu. (6) For a large gamma, magnitude of mu decreases with the thickness of membrane d under the condition of constant amount of fixed charges. However, if gamma is sufficiently small, the variation of magnitude of mu as a function of d exhibits a maximum. The classic result of Smoluchowski for the electrophoretic mobility of a rigid particle can be recovered as a limiting case of the present model.

Animals↗
PubMed1972

The ionogenic nature of the secretory-granule membrane. Electrokinetic properties of isolated chromaffin granules.

1. Chromaffin granules isolated from the bovine adrenal medulla possess an electrophoretic mobility of -1.12mum.s(-1).cm.V(-1), corresponding to a surface zeta potential of -14.4mV and surface charge density of 1.38x10(-6)C.cm(-2). 2. The mobility of chromaffin granules is pH-dependent, indicating an amphoteric surface with an isoelectric point at pH3.0 and acidic groups with a pK(a) of 3.11. 3. Addition of bi- and ter-valent cations decreased the mobility of chromaffin granules in a dose-dependent fashion with a relative potency of La(3+)>>Mn(2+)>Ca(2+) >Sr(2+)>Mg(2+)>Ba(2+). 4. Treatment with neuraminidase decreased the mobility of erythrocytes by 84%, whereas chromaffin-granule mobility was decreased by only 14%. This correlates well with the small complement of neuraminic acid present in the granule membrane. 5. The nature, origin and significance of the anionic surface charge of the chromaffin granule is discussed. It is concluded that the net negative charge at the surface of shear derives chiefly from a single type of chemical group, namely -CO(2) (-), contributed by the alpha-carboxyl group of constituent proteins, the phospholipid phosphatidylserine and, to a lesser extent, the sialic acid component of glycoproteins.

Adrenal Medulla↗
PubMed2001

Dipole potentials indicate restructuring of the membrane interface induced by gadolinium and beryllium ions.

The dipole component of the membrane boundary potential, phi(d), is an integral parameter that may report on the conformational state of the lipid headgroups and their hydration. In this work, we describe an experimental approach to measurements of the dipole potential changes, Deltaphi(d), and apply it in studies of Be(2+) and Gd(3+) interactions with membranes composed of phosphatidylserine (PS), phosphatidylcholine (PC), and their mixtures. Deltaphi(d) is determined as the difference between the changes of the total boundary potential, phi(b), measured by the IFC method in planar lipid membranes and the surface potential, phi(s), determined from the electrophoretic mobility of liposomes. The Gouy-Chapman-Stern formalism, combined with the condition of mass balance, well describes the ion equilibria for these high-affinity cations. For the adsorption of Be(2+) and Gd(3+) to PC membranes, and of Mg(2+) to PS membranes, the values of Deltaphi(b) and Deltaphi(s) are the same, indicative of no change of phi(d). Binding of Gd(3+) to PS-containing membranes induces changes of phi(d) of opposite signs depending on the density of ionized PS headgroups in the bilayer. At low density, the induced Deltaphi(d) is negative (-30 mV), consistent with the effect of dehydration of the surface. At maximal density (pure PS, neutral pH), adsorption of Be(2+) or Gd(3+) induces an increase of phi(d) of 35 or 140 mV, respectively. The onset of the strong positive dipole effect on PS membranes with Gd(3+) is observed near the zero charge point and correlates with a six-fold increase of membrane tension. The observed phenomena may reflect concerted reorientation of dipole moments of PS headgroups as a result of ion adsorption and lipid condensation. Their possible implications to in-vivo effects of these high-affinity ions are discussed.

Adsorption↗
PubMed2003

Electrosurface phenomena at polymer films for biosensor applications.

Electrosurface phenomena at thin polymer films utilized in the preparation of biosensors have been studied by use of the Microslit Electrokinetic set-up. For the investigated polymer layers (aminodextran, carboxylated dextran, diaminopoly(ethylene glycol), dicarboxypoly(ethylene glycol), biotinylated poly(ethylene glycol), and streptavidin on biotinylated poly(ethylene glycol), the charge formation in aqueous electrolyte solutions was found to depend on the pH value, that is, OH- and H3O+ are the charge determining ions. The isoelectric points obtained from zeta potential versus pH plots could be utilized to draw conclusions on the introduction of acidic or basic groups and on the degree of molecular surface coverage, respectively. The hydrodynamically mobile charge reflected by the magnitude of the zeta potential contributed to only about 6% or less of the total surface conductivity of the polymer layers. The experimental determination of the total surface conductivity was found to provide valuable information on structural features of biosensor interfaces in aqueous environments.

Biosensing Techniques↗
PubMed2002

Efficient mobilization of E1-deleted adenovirus type 5 vectors by wild-type adenoviruses of other serotypes.

Mobilization of replication-deficient adenovirus vectors can lead to spread and shedding of the vector. Here we show that in cultured HepG2 cells wild-type (wt) adenoviruses of subgroup A (Ad12), B (Ad7, 11 and 16), C (Ad1, 2 and 5) and E (Ad4) can efficiently mobilize Ad5CMVluc, a DeltaE1DeltaE3-Ad5 vector carrying the firefly luciferase gene as reporter. In addition, we show that Ad5CMVluc can be propagated on Ad12E1-transformed human embryonic retinoblasts. This provides evidence that expression of the E1 region of Ad12 is sufficient for mobilizing DeltaE1-Ad5-derived vectors. Thus, in therapeutic applications of replication-defective Ad vectors any active Ad infection is of potential concern, independent of the serotype involved. To prevent vector mobilization by wt Ads, new vectors should be developed in which essential functions such as the initiation of DNA replication and genome packaging are restricted.

Adenovirus E1 Proteins↗
PubMedDate not supplied

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↗
PubMed1983

Large divalent cations and electrostatic potentials adjacent to membranes. Experimental results with hexamethonium.

A simple extension of the Gouy-Chapman theory predicts that the ability of a divalent cation to screen charges at a membrane-solution interface decreases significantly if the distance between the charges on the cation is comparable with the Debye length. We tested this prediction by investigating the effect of hexamethonium on the electrostatic potential adjacent to negatively charged phospholipid bilayer membranes. The distance between the two charges of an extended hexamethonium molecule is approximately 1 nm, which is the Debye length in the 0.1 M monovalent salt solutions used in these experiments. Six different experimental approaches were utilized. We measured the electrophoretic mobility of multilamellar vesicles to determine the zeta potential, the line width of the 31P nuclear magnetic resonance (NMR) signal from sonicated vesicles to calculate the change in potential at the phosphodiester moiety of the lipid, and the conductance of planar bilayer membranes exposed to either carriers (nonactin) or pore formers (gramicidin) to estimate the change in potential within the membrane. We also measured directly the effect of hexamethonium on the potential above a monolayer formed from negative lipids, and attempted to calculate the change in the surface potential of a bilayer membrane from capacitance measurements. With the exception of the capacitance calculations, each of the techniques gave comparable results: hexamethonium exerts a smaller effect on the potential than that predicted by the classic screening theory. The results are consistent with the predictions of the extended Gouy-Chapman theory and are relevant to the interpretation of physiological and pharmacological experiments that utilize hexamethonium and other large divalent cations.

Animals↗
PubMed2002

Use of nonionic poly(ethylene glycol) p-isooctyl-phenyl ether (Triton X-1 00) surfactant mobile phases in the thin-layer chromatography of heavy-metal cations.

The analytical potential of poly(ethylene glycol) p-isooctyl-phenyl ether (Triton X-100), a nonionic surfactant, is used as a mobile phase in the thin-layer chromatographic separation of heavy-metal cations. The surfactant concentration below its critical micellar concentration (CMC) as well as above the CMC value is used to investigate the migrational behavior of some heavy-metal ions on silica gel layers. The mobility of the metal ions is found to change marginally with the increase of surfactant concentration from 0.001M (below CMC) to 0.1M (above CMC). The influence of the pH of the medium, nonelectrolyte organic (urea and alkanols), and inorganic electrolyte (NaCI) additives in the surfactant containing mobile phase on the mobility of heavy metals on the silica gel layer is examined. For separating metal ions, surfactant must be used in the presence of buffers. Triton X-100 (0.02M) at pH 2.3 is found to be the best mobile phase for the separation of heavy-metal cations. In general, the presence of alcohol in aqueous surfactant solutions results in a decrease in the mobility of metal ions. Besides Cu2+ and Fe3+, all of the metal ions show a trend of increasing the retardation factor beyond a minima at 0.1 or 0.3M of added urea or NaCl. The proposed method is successfully applied for the simultaneous detection of Zn2+ and Cd2+ from a spiked human blood sample.

Journal Article↗
PubMed1976

The effect of chelating agents on iron mobilization in Chang cell cultures.

The investigation of chelating agents with potential therapeutic value in patients with transfusional iron overload has been facilitated by the use of Chang cell cultures. These cells have been incubated with [59Fe]transferrin for 22 hr, following which most of the intracellular radioiron is found in the cytosol, distributed between a ferritin and a nonferritin form. Iron release from the cells depends on transferrin saturation in the medium, but when transferrin is 100% saturated, which normally does not allow iron release, desferrioxamine, 2,3-dihydroxybenzoic acid, rhodotorulic acid, cholythydroxamic acid, and tropolone all promote the mobilization of ferritin iron and its release from cells. They are effective to an approximately equal degree. The incubation of [59Fe]transferrin with tropolone in vitro at a molar ratio of 1:500 results in the transfer of most of the labeled iron to the chelator, reflecting the exceptionally high binding constant of this compound. How far these phenomena relate to therapeutic potentially remains to be seen.

Anemia, Hypochromic↗
PubMedDate not supplied

Is there evidence for early mobilization following an extraarticular hand fracture?

The authors conducted a systematic review to determine if there is scientifically valid (level I or II) evidence for the effect of early motion (<21 days) of joints surrounding an extraarticular hand fracture on fracture healing or functional outcomes. Two reviewers independently evaluated for study inclusion, trial quality and internal validity. Six poor-quality, quasirandomized studies (level III evidence) involving 459 patients were included. Findings suggest that in simple, closed metacarpal fractures, early motion (EM) has the potential to: (1) result in earlier recovery of mobility and strength, (2) facilitate an earlier return to work, and (3) not affect fracture alignment. Findings also indicate that skin pressure problems are not associated with custom-molded metacarpal fracture braces. The scientific validity of EM interventions after an extraarticular hand fracture has not been established in well-conducted, randomized, controlled trials (level I or II evidence). Current evidence does not support or refute the use of EM after an extraarticular hand fracture. However, further investigations are warranted, as findings to date show a consistent potential for benefit with no significant risk of harm when early regional joint motions are incorporated into the management of closed, extraarticular, finger metacarpal fractures.

Early Ambulation↗