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PubMed1988

Characterization of guinea-pig cardiac muscarinic receptors by radioligand dissociation kinetics.

The question of cardiac muscarinic receptor heterogeneity was studied in guinea-pig auricles and ventricle. Radioligand dissociation kinetics were analyzed in the absence and presence of alinidine and AQ-A 39, two muscarinic agents and allosteric modulators of radiotracer dissociation. The dissociation kinetics were monophasic with all 3 radiotracers used and in both auricles and ventricle. [125I]3-Quinuclidinyl 4-iodobenzilate ([125I]QNB) dissociated with an identical half-life (t1/2 off) in ventricle and in left and right auricle, respectively. Alinidine (1 mM) decreased t1/2 off by 50% in ventricle but had no significant influence in auricles. AQ-A 39 (= falipamil) uniformly increased t1/2 off 4-fold in all 3 tissues. The binding of tritium-labelled quinuclidinyl benzilate ([3H]QNB) dissociated equally rapidly from both ventricular and auricular receptors under control conditions but was not affected by alinidine. AQ-A 39 (1 mM) slowed [3H]QNB dissociation 6.5-fold in ventricle but only 3-fold in both auricles. [3H]N-Methyl scopolamine ([3H]NMS) dissociation was uniform in both auricles but was somewhat slower in the ventricle, both in the absence and presence of alinidine. AQ-A 39 was without effect. These results demonstrate differences in ventricular versus auricular receptors detected by [125I]QNB in the presence of alinidine and by [3H]QNB in the presence of AQ-A 39. The quaternary ligand [3H]NMS was unable to detect receptor heterogeneity. No differences were found between right and left auricular receptors. The results presented provide no evidence for the existence of different subtypes of muscarinic receptors for the negative chronotropic and inotropic actions of muscarinic agonists.

Allosteric Site↗
PubMed1989

Dissociation pathway for recA nucleoprotein filaments formed on linear duplex DNA.

recA protein forms stable filaments on duplex DNA at low pH. When the pH is shifted above 6.8, recA protein remains stably bound to nicked circular DNA, but not to linear DNA. Dissociation of recA protein from linear duplex DNA proceeds to a non-zero endpoint. The kinetics and final extent of dissociation vary with several experimental parameters. The instability on linear DNA is most readily explained by a progressive unidirectional dissociation of recA protein from one end of the filament. Dissociation of recA protein from random points in the filament is eliminated as a possible mechanism by several observations: (1) the requirement for a free end; (2) the inverse and linear dependence of the rate of dissociation on DNA length (at constant DNA base-pair concentration); and (3) the kinetics of exposure of a restriction endonuclease site in the middle of the DNA. Evidence against another possible mechanism, ATP-mediated translocation of the filament along the DNA, is provided by a novel effect of the non-hydrolyzable ATP analog, ATP gamma S, which generally induces recA protein to bind any DNA tightly and completely inhibits ATP hydrolysis. We find that very low, sub-saturating levels of ATP gamma S completely stabilize the filament, while most of the ATP hydrolysis continues. If these levels of ATP gamma S are introduced after dissociation has commenced, further dissociation is blocked, but re-association does not occur. These observations are inconsistent with movement of recA protein along DNA that is tightly coupled to ATP hydrolysis. The recA nucleoprotein filament is polar and the protein binds the two strands asymmetrically, polymerizing mainly in the 5' to 3' direction on the initiating strand of a single-stranded DNA tailed duplex molecule. A model consistent with these results is presented.

Adenosine Triphosphate↗
PubMed1983

The dissociation of testosterone- and 5 alpha-dihydrotestosterone-receptor complexes formed within cultured human genital skin fibroblasts.

We have studied the rate and character with which testosterone (T) and 5 alpha-dihydrotestosterone (DHT) dissociate from the androgen receptor both within intact cultured genital skin fibroblasts of a subject with 5 alpha-reductase deficiency and after the androgen-receptor complexes have been extracted from the cells. Within the cells, the kinetics of the dissociative process for each hormone was first-order, but T dissociated four times faster than DHT. An Arrhenius plot of the variation of the dissociation rate constants with temperature for T was linear and yielded an activation energy of 28 kcal/mol. This value is identical with the one previously determined for activated DHT-receptor complexes. T-receptor complexes extracted from the cells dissociated with complex kinetics: at 37 degrees C the rate constants of the "fast" and "slow" components were 40 and 14 X 10(-3) min-1, respectively. In contrast, DHT-receptor complexes extracted from the cells dissociated with first-order kinetics and at a rate identical to that observed within cells, except after exposure to pyridoxal 5'-phosphate (5 mM) or concentration by Amicon (B-15) filtration, when their dissociation kinetics became complex. We interpret these data to mean that, within the cells, both T- and DHT-receptor complexes exist predominantly in the activated state whereas, when extracted from the cells, DHT-receptor complexes remain activated, unless perturbed, while T-receptor complexes become unstable spontaneously, probably by reverting to a preactivated state.

Cells, Cultured↗
PubMed1985

A method for in vitro enzymatic dissociation of nerve roots and peripheral nerves from adult mammals.

Preparations yielding a high percentage of undamaged axons from fresh peripheral nerve or nerve root were made using an enzymatic dissociation regimen. The nerve was placed in a temperature-controlled chamber mounted over an inverted phase-contrast microscope. An oxygenated solution (Brimijoins) or modified Hank's solution was pumped through the chamber, first in a calcium-free form and then containing enzymes. The enzymes for dissociation were collagenase and trypsin, alternated. Enzymatic dissociation of the epineurium, perineurium and extracellular matrix was achieved. We supplemented the gentle agitation of a 10-roller peristaltic pump by periodically raising and lowering the fluid level in the chamber to provide a controlled mechanical agitation that promoted dissociation. A large percentage of the axons can be dissociated from the nerve, varying from approximately one-quarter to occasional complete dissociation. Action potentials were still conducted through dissociated axons, and axon transport was also still present, as documented by direct visualization using an AVEC-DIC type of microscope system. The axons had a better morphological appearance and displayed better transport than comparison preparations prepared by the usual mechanical teasing method, in our hands. The enzymatic method allows study of axons in an adult or developing mammal with regard to their electrical conduction and axon transport mechanisms. It should help to avoid a selection process for more hardy axons which may be imposed by traditional mechanical teasing methods. Mechanical stress was observed to cause widened Schmidt-Lanterman clefts, widened nodes, myelin bubbles, and other abnormal morphology as evidence of damage.

Animals↗
PubMed1985

The rate of MgADP binding to and dissociation from acto-S1.

The rate of binding and dissociation of MgADP from its ternary complex with actin and S1 was measured by following the extent to which fixed concentrations of MgADP slow down MgATP-induced dissociation of acto-S1. The solution of the equations describing this process shows that at any MgADP concentration the apparent rate of acto-S1 dissociation should be proportional to a square root of the equilibrium constant for MgADP dissociation and to MgATP concentration. By measuring the apparent rate of acto-S1 dissociation as a function of MgATP concentration, the rate of MgADP binding and dissociation were determined as 5 X 10(6) M-1 X s-1 and 1400 s-1, respectively. These rates were unchanged by modification of SH1 thiol of S1 by a variety of fluorescence and spin-labels, but dissociation rate was drastically reduced when SH1 was labelled with 5-iodoacetamidofluorescein.

Actins↗
PubMed1987

Hydrophobic stabilization of chiton hemocyanins: effects of ureas, Hofmeister salts and pH on their dissociation.

The subunit dissociation of the hemocyanins from five members of the Polyplacophora families, Acanthochitonidae, Callistoplacidae, Chitonidae, Ischnochitonidae and Mopalidae, represented by the chitons Cryptochiton stelleri, Nutallina fluxa, Acanthopleura granulata, Stenoplax conspicua and Mopalia mucosa, respectively, have been investigated by light-scattering molecular-weight and ultracentrifugation methods, using the hydrophobic reagents of the urea series and the Hofmeister salt series as probes of the contact areas of the hemocyanin subunits. The polyplacophoran hemocyanins are decamers with molecular weights of (4.2-4.6) X 10(6). The effectiveness of dissociation by the ureas follows the order of increasing hydrophobicity of the reagent, i.e., urea, methyl-, ethyl-, propyl- and butylurea, as expected of hydrophobically stabilized subunit systems. The urea dissociation is found to be a two-step reaction: the dissociation of parent decamers to dimers followed by dissociation of the dimers to monomers. Analysis of the observed decrease in molecular weight requires the interaction with urea of about 27 to 35 apparent amino-acid groups (Napp) at the contact areas of the dimers, and a much larger number of apparent binding groups ranging from about 100 to 120 per monomer at the contact areas of the monomers. Fitting of the pH dissociation profiles of A. granulata, C. stelleri, M. muscosa and S. conspicua requires the participation of a much smaller number of amino-acid residues in the interaction with the probe-solvent components. The ionization or protonation of one acidic and one basic group per dimer, and five to eight acidic and basic groups per monomer is found to be adequate for the description of the two-step pH dissociation reaction.

Animals↗
PubMed1976

The dissociation of insulin from human insulin antibodies.

The dissociation of insulin from human insulin antibodies has been investigated using a technique that is rapid and does not require addition of excess unlabelled insulin. A slow (k1 = 2-10(-3) min-1) and a fast k2 = 4-10(-2) min-1) dissociating antibody component were identified in all studies. These have been shown to correspond, respectively, to the high and low affinity antibody components of equilibrium binding studies. The range of k1 and k2 values and their response to temperature change is small. Insulin resistance and stability of diabetes are not related to properties of antibody dissociation. Dissociation is faster in the presence of high (6-850 nM) insulin concentration due to increased binding to the fast dissociating component without change in the dissociation rate constants. When incubation time is increased beyond achievement of maximal binding there is a time-depent rise in binding to the slow dissociating component, with a concomitant fall in k1. The traditional concept that equilibrium is established at maximum binding requires further examination.

Antigen-Antibody Reactions↗
PubMed2006

The role of depression and dissociation in the relationship between childhood trauma and bulimic symptoms among ethnically diverse female undergraduates.

OBJECTIVE: The goals of this study were to examine the role of dissociation and depression as possible mediators of the relationship between several forms of childhood trauma and bulimic symptomatology and to explore potential ethnic differences in these relationships. METHOD: Four hundred seventeen female undergraduates participated in this cross-sectional study. They completed measures of dissociative, depressive, and bulimic symptoms, and childhood trauma. Experiences of multiple forms of childhood trauma were measured, including physical abuse, sexual abuse, emotional abuse, physical neglect, and emotional neglect. However, only emotional abuse was correlated with bulimic symptoms at p<.01. Therefore, other forms of trauma were excluded from the analyses to control for Type I error. RESULTS: Dissociation was not associated with emotional abuse after controlling for depression; therefore, tests of dissociation as a mediator were discontinued. Depression was significantly associated with emotional abuse after controlling for dissociation. Emotional abuse was significantly associated with bulimia. Finally, emotional abuse and depression together were significantly associated with bulimia after controlling for dissociation. However, emotional abuse became nonsignificant when entered with depression, indicating that depression mediated the relationship between emotional abuse and bulimic symptoms. There were no ethnic differences in this relationship. CONCLUSIONS: It appears that among women who have experienced childhood emotional abuse, depression is more strongly associated with unhealthy eating behaviors than is dissociation. Results also suggest that emotional abuse is a form of childhood trauma particularly relevant to bulimia.

Adolescent↗
PubMed2005

Rates of nitric oxide dissociation from hemoglobin.

Nitric oxide (NO) plays a major role in human physiology and in many pathological states. Although oxyhemoglobin is known to destroy NO activity, NO activity can, in principle, be conserved through iron nitrosylation at vacant hemes. In order for this NO activity to be delivered, the NO must dissociate from the heme. Despite its study over the past few decades, our understanding of NO dissociation from hemoglobin is incomplete. In principle, there are at least four NO dissociation rates: kR(alpha), kR(beta), kT(alpha), and kT(beta), where the subscript refers to the quaternary state and the superscript to the hemoglobin chain. In the T-state, a proportion of the proximal histidine bonds break forming pentacoordinate alpha-nitrosyl hemoglobin. In vivo, alpha-nitrosyl hemoglobin predominates over beta-nitrosyl hemoglobin. In this study we have used a fast NO trap, Fe(II)-proline-dithiocarbamate, to measure NO dissociation rates from hemoglobin. We have varied solution conditions so the rate of dissociation from pentacoordinate alpha-nitrosyl hemoglobin could be definitively measured for the first time; kT(alpha) = 4.2 +/- 1.5 x 10(-4) s(-1). We have also found that the fastest NO dissociation rate is on the order of 10(-3) s(-1) and that NO dissociation from sickle cell hemoglobin is the same as that from normal adult hemoglobin.

Electron Spin Resonance Spectroscopy↗
PubMed2004

Modeling deoxyribose radicals by neutralization-reionization mass spectrometry. Part 2. Preparation, dissociations, and energetics of 3-hydroxyoxolan-3-yl radical and cation.

The title radical (1) is generated in the gas-phase by collisional neutralization of carbonyl-protonated oxolan-3-one. A 1.5% fraction of 1 does not dissociate and is detected following reionization as survivor ions. The major dissociation of 1 (approximately 56%) occurs as loss of the hydroxyl H atom forming oxolan-3-one (2). The competing ring cleavages by O[bond]C-2 and C-4[bond]C-5 bond dissociations combined account for approximately 42% of dissociation and result in the formation of formaldehyde and 2-hydroxyallyl radical. Additional ring-cleavage dissociations of 1 resulting in the formation of C(2)H(3)O and C(2)H(4)O cannot be explained as occurring competitively on the doublet ground (X) electronic state of 1, but are energetically accessible from the A and higher electronic states accessed by vertical electron transfer. Exothermic protonation of 2 also produces 3-oxo-(1H)-oxolanium cation (3(+)) which upon collisional neutralization gives hypervalent 3-oxo-(1H)-oxolanium radical (3). The latter dissociates spontaneously by ring opening and expulsion of hydroxy radical. Experiment and calculations suggest that carbohydrate radicals incorporating the 3-hydroxyoxolan-3-yl motif will prefer ring-cleavage dissociations at low internal energies or upon photoexcitation by absorbing light at approximately 590 and approximately 400 nm.

Cations↗
PubMed2005

Toward a general mechanism of electron capture dissociation.

The effects of positive charge on the properties of ammonium and amide radicals were investigated by ab initio and density functional theory calculations with the goal of elucidating the energetics of electron capture dissociation (ECD) of multiply charged peptide ions. The electronic properties of the amide group in N-methylacetamide (NMA) are greatly affected by the presence of a remote charge in the form of a point charge, methylammonium, or guanidinium cations. The common effect of the remote charge is an increase of the electron affinity of the amide group, resulting in exothermic electron capture. The N-Calpha bond dissociation and transition state energies in charge-stabilized NMA anions are 20-50 kJ mol(-1) greater than in the hydrogen atom adduct. The zwitterions formed by electron capture have proton affinities that were calculated as 1030-1350 kJ mol(-1), and are sufficiently basic for the amide carbonyl to exothermically abstract a proton from the ammonium, guanidinium and imidazolium groups in protonated lysine, arginine, and histidine residues, respectively. A new mechanism is proposed for ECD of multiply charged peptide and protein cations in which the electron enters a charge-stabilized electronic state delocalized over the amide group, which is a superbase that abstracts a proton from a sterically proximate amino acid residue to form a labile aminoketyl radical that dissociates by N-Calpha bond cleavage. This mechanism explains the low selectivity of N-Calpha bond dissociations induced by electron capture, and is applicable to dissociations of peptide ions in which the charge carriers are metal ions or quaternary ammonium groups. The new amide superbase and the previously proposed mechanisms of ECD can be uniformly viewed as being triggered by intramolecular proton transfer in charge-reduced amide cation-radicals. In contrast, remote charge affects N-H bond dissociation in weakly bound ground electronic states of hypervalent ammonium radicals, as represented by methylammonium, CH3NH3*, but has a negligible effect on the N-H bond dissociation in the strongly bound excited electronic states. This refutes previous speculations that loss of "hot hydrogen" can occur from an excited state of an ammonium radical.

Acetamides↗
PubMed2005

Gas-phase dissociation of oligoribonucleotides and their analogs studied by electrospray ionization tandem mass spectrometry.

Oligoribonucleotides (RNA) and modified oligonucleotides were subjected to low-energy collision-induced dissociation in a hybrid quadrupole time-of-flight mass spectrometer to investigate their fragmentation pathways. Only very restricted data are available on gas-phase dissociation of oligoribonucleotides and their analogs and the fundamental mechanistic aspects still need to be defined to develop mass spectrometry-based protocols for sequence identification. Such methods are needed, because chemically modified oligonucleotides can not be submitted to standard sequencing protocols. In contrast to the dissociation of DNA, dissociation of RNA was found to be independent of nucleobase loss and it is characterized by cleavage of the 5'-P-O bond, resulting in the formation of c- and their complementary y-type ions. To evaluate the influence of different 2'-substituents, several modified tetraribonucleotides were analyzed. Oligoribonucleotides incorporating a 2'-methoxy-ribose or a 2'-fluoro-ribose show fragmentation that does not exhibit any preferred dissociation pathway because all different types of fragment ions are generated with comparable abundance. To analyze the role of the nucleobases in the fragmentation of the phosphodiester backbone, an oligonucleotide lacking the nucleobase at one position has been studied. Experiments indicated that the dissociation mechanism of RNA is not influenced by the nucleobase, thus, supporting a mechanism where dissociation is initiated by formation of an intramolecular cyclic transition state with the 2'-hydroxyl proton bridged to the 5'-phosphate oxygen.

DNA↗
PubMed2005

Blackbody infrared radiative dissociation of nonspecific protein-carbohydrate complexes produced by nanoelectrospray ionization: the nature of the noncovalent interactions.

Gas-phase thermal dissociation experiments, implemented with blackbody infrared radiative dissociation (BIRD) and Fourier transform ion cyclotron resonance mass spectrometry, have been performed on a series of protonated and deprotonated 1:1 and protonated 1:2 protein-carbohydrate complexes formed by nonspecific interactions during the nanoflow electrospray (nanoES) ionization process. Nonspecific interactions between the proteins bovine carbonic anhydrase II (CA), bovine ubiquitin (Ubq), and bovine pancreatic trypsin inhibitor and several carbohydrates, ranging in size from mono- to tetrasaccharides, have been investigated. Over the range of temperatures studied (60-190 degrees C), BIRD of the protonated and deprotonated complexes proceeds exclusively by the loss of the carbohydrate in its neutral form. The rates of dissociation of the 1:1 complexes containing a mono- or disaccharide decrease with reaction time, suggesting the presence of two or more kinetically distinct structures produced during nanoES or by gas-phase processes. In contrast, the 1:1 complexes of the tri- and tetrasaccharides exhibit simple first-order dissociation kinetics, a result that, on its own, is suggestive of a single preferred carbohydrate binding site or multiple equivalent sites in the gas phase. A comparative analysis of the dissociation kinetics measured for protonated 1:1 and 1:2 complexes of Ubq with alphaTal[alphaAbe]alphaMan further supports the presence of a single preferred binding site. However, a similar analysis performed on the complexes of CA and alphaTal[alphaAbe]alphaMan suggests that equivalent but dependent carbohydrate binding sites exist in the gas phase. Analysis of the Arrhenius activation parameters (E(a) and A) determined for the dissociation of 1:1 complexes of CA with structurally related trisaccharides provides evidence that neutral intermolecular hydrogen bonds contribute, at least in part, to the stability of the gaseous complexes. Surprisingly, the E(a) values for the complexes of the same charge state are not sensitive to the structure (primary or higher order) of the protein, suggesting that the carbohydrates are able to form energetically equivalent interactions with the various functional groups presented by the protein. For a given protein-carbohydrate complex, the dissociation E(a) is sensitive to charge state, initially increasing and then decreasing with increasing charge. It is proposed that both ionic and neutral hydrogen bonds stabilize the nonspecific protein-carbohydrate complexes in the gas phase and that the relative contribution of the neutral and ionic interactions is strongly influenced by the charge state of the complex, with neutral interactions dominating at low charge states and ionic interactions dominating at high charge states.

Binding Sites↗
PubMed2007

Influence of Coulombic repulsion on the dissociation pathways and energetics of multiprotein complexes in the gas phase.

Thermal dissociation experiments, implemented with blackbody infrared radiative dissociation and Fourier-transform ion cyclotron resonance mass spectrometry, are performed on gaseous protonated and deprotonated ions of the homopentameric B subunits of Shiga toxin 1 (Stx1 B5) and Shiga toxin 2 (Stx2 B5) and the homotetramer streptavidin (S4). Dissociation of the gaseous, multisubunit complexes proceeds predominantly by the loss of a single subunit. Notably, the fractional partitioning of charge between the product ions, i.e., the leaving subunit and the resulting multimer, for a given complex is, within error, constant over the range of charge states investigated. The Arrhenius activation parameters (E(a), A) measured for the loss of subunit decrease with increasing charge state of the complex. However, the parameters for the protonated and deprotonated ions, with the same number of charges, are indistinguishable. The influence of the complex charge state on the dissociation pathways and the magnitude of the dissociation E(a) are modeled theoretically with the discrete charge droplet model (DCDM) and the protein structure model (PSM), wherein the structure of the subunits is considered. Importantly, the major subunit charge states observed experimentally for the Stx1 B5(n+/-) ions correspond to the minimum energy charge distribution predicted by DCDM and PSM assuming a late dissociative transition-state (TS); while for structurally-related Stx2 B5(n+) ions, the experimental charge distribution corresponds to an early TS. It is proposed that the lateness of the TS is related, in part, to the degree of unfolding of the leaving subunit, with Stx1 B being more unfolded than Stx2 B. PSM, incorporating significant subunit unfolding is necessary to account for the product ions observed for the S4(n+) ions. The contribution of Coulombic repulsion to the dissociation E(a) is quantified and the intrinsic activation energy is estimated for the first time.

Gases↗
PubMed2005

Childhood stuttering and dissociations across linguistic domains.

UNLABELLED: The purpose of this investigation was to evaluate the possible presence of dissociations in the speech and language skills of young children who do (CWS) and do not stutter (CWNS) using a correlation-based statistical procedure [Bates, E., Appelbaum, M., Salcedo, J., Saygin, A. P., & Pizzamiglio, L. (2003). Quantifying dissociations in neuropsychological research. Journal of Clinical and Experimental Neuropsychology, 25, 1128-1153]. Participants were 45 preschool CWS and 45 CWNS between the ages of 3;0 and 5;11 (years;months), with the two groups matched by age, gender, race, and parental socioeconomic status. Children participated in a parent-child interaction for the purpose of disfluency analysis and responded to four standardized speech-language tests for subsequent analyses as main dependent variables. Findings indicated that CWS were over three times more likely than CWNS to exhibit dissociations across speech-language domains, with 44 cases of dissociation for CWS and 14 for CWNS across 10 possible comparisons. Results suggest that there may be a subgroup of CWS who exhibit dissociations across speech-language domains, which may result in a greater susceptibility to breakdowns in speech fluency. EDUCATIONAL OBJECTIVES: The reader will be able to: (1) summarize findings from previous studies examining differences in speech and language performance between children who do and do not stutter; (2) describe what is meant by "dissociations" in the speech and language skills of young children who do and do not stutter; and (3) discuss three hypotheses that could account for the present findings that suggest CWS, more often than CWNS, exhibit dissociations in their speech-language system.

Child↗
PubMed2006

Methods for isolating highly-enriched embryonic spinal cord neurons: a comparison between enzymatic and mechanical dissociations.

Spinal cord neuronal culture is a useful system to study normal and abnormal functions of the spinal cord. For many bioassays, obtaining large quantities of highly purified spinal cord neurons is required. However, technical difficulties exist in obtaining these cells reliably and consistently. By comparing two dissociation methods, mechanical and enzymatic dissociations, we found that the enzymatic dissociation of embryonic day 14-15 spinal cords resulted in significantly higher cell yield than the mechanical dissociation (25.40 +/- 5.41 x 10(6) versus 3.43 +/- 0.52 x 10(6) cells per 12 embryos; n = 6/group; p < 0.01). Furthermore, cell viability was significantly higher after the enzymatic than the mechanical dissociation (83.40 +/- 3.08% versus 32.81 +/- 3.49%, n = 4/group; p < 0.01). In both methods, highly purified populations of primary neurons were obtained (mechanical: 85.17 +/- 2.84%; enzymatic: 87.67 +/- 2.52%; n = 3/group). Critical measures that affect culture outcomes include, but not limited to, the age of embryo, cell seeding density, dissociation time, and elimination of non-neuronal cells. Thus, the present study has identified the enzymatic dissociation method to be a preferred method for obtaining large quantity of highly-enriched embryonic spinal cord neurons.

Age Factors↗
PubMed1995

The prevalence of dissociated vertical deviation in patients with sensory heterotropia.

PURPOSE: We reviewed 281 consecutive patients with sensory esotropia or exotropia to determine the prevalence of dissociated vertical deviation in patients with sensory strabismus. METHODS: We reviewed the charts of all patients who received a diagnosis of sensory heterotropia or dissociated vertical deviation and who were examined at the Pediatric Ophthalmology Service at the Texas Children's Hospital between 1973 and 1992. Statistical analyses of the prevalence of dissociated vertical deviation were determined after evaluating the direction of the accompanying horizontal strabismus and examining the temporal relationship of the unilateral vision loss. RESULTS: Dissociated vertical deviation was diagnosed in 35 patients (12.5%). It occurred more frequently in sensory esotropia (22 patients, 18.3%) than exotropia (13 patients, 8.1%) (P = .009). The age at which unilateral visual loss occurred did not influence the development of dissociated vertical deviation. CONCLUSION: Dissociated vertical deviation in patients with acquired loss of vision does not support the contention that dissociated vertical deviation is a genetically predetermined anomaly of binocular vision.

Adolescent↗
PubMed1989

Competition between solution and cell surface receptors for ligand. Dissociation of hapten bound to surface antibody in the presence of solution antibody.

We present a joint theoretical and experimental study on the effects of competition for ligand between receptors in solution and receptors on cell surfaces. We focus on the following experiment. After ligand and cell surface receptors equilibrate, solution receptors are introduced, and the dissociation of surface bound ligand is monitored. We derive theoretical expressions for the dissociation rate and compare with experiment. In a standard dissociation experiment (no solution receptors present) dissociation may be slowed by rebinding, i.e., at high receptor densities a ligand that dissociates from one receptor may rebind to other receptors before separating from the cell. Our theory predicts that rebinding will be prevented when S much greater than N2Kon/(16 pi 2D a4), where S is the free receptor site concentration in solution, N the number of free surface receptor sites per cell, Kon the forward rate constant for ligand-receptor binding in solution, D the diffusion coefficient of the ligand, and a the cell radius. The predicted concentration of solution receptors needed to prevent rebinding is proportional to the square of the cell surface receptor density. The experimental system used in these studies consists of a monovalent ligand, 2,4-dinitrophenyl (DNP)-aminocaproyl-L-tyrosine (DCT), that reversibly binds to a monoclonal anti-DNP immunoglobulin E (IgE). This IgE is both a solution receptor and, when anchored to its high affinity Fc epsilon receptor on rat basophilic leukemia (RBL) cells, a surface receptor. For RBL cells with 6 x 10(5) binding sites per cell, our theory predicts that to prevent DCT rebinding to cell surface IgE during dissociation requires S much greater than 2,400 nM. We show that for S = 200-1,700 nM, the dissociation rate of DCT from surface IgE is substantially slower than from solution IgE where no rebinding occurs. Other predictions are also tested and shown to be consistent with experiment.

Animals↗