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PubMedDate not supplied

[Effect of carbon, nitrogen, and phosphorus sources of nutrition on the growth of R-, S- and M-dissociants of Pseudomonas aeruginosa].

The effect of glucose, nitrate, and phosphate on the stationary-phase growth parameters of the R, S, and M dissociants of the hydrocarbon-oxidizing bacterium Pseudomonas aeruginosa K-2 was studied. The data obtained were analyzed in terms of the Mitscherlich equation. S dissociant required less glucose than other dissociants, whereas M dissociant required less nitrogen and phosphorus. These findings were confirmed by the results of investigation of the combined action of glucose, nitrate, and phosphate in a 3 x 3 x 3 factor experiment. It is anticipated that M dissociant must prevail under conditions of nitrogen and phosphorus deficiency, and S dissociant must be dominant in the case of optimally chosen proportions between the biogenic elements studied.

Carbon↗
PubMed1999

Dissociated glucocorticoids with anti-inflammatory potential repress interleukin-6 gene expression by a nuclear factor-kappaB-dependent mechanism.

Synthetic glucocorticoids (GCs) remain among the most effective agents for the management of chronic inflammatory diseases. However, major side effects severely limit their therapeutic use. Physiologic and therapeutic activities of GCs are mediated by a nuclear receptor belonging to a superfamily of ligand-inducible transcription factors that, in addition to directly regulating their cognate gene programs, can also mutually interfere with other signaling pathways. We recently identified selective ligands of the glucocorticoid receptor that dissociate transactivation from activator protein 1 transrepression, and most importantly retain in vivo anti-inflammatory activity. To further document the mechanisms of action sustaining the observed in vivo activity, we report here on the interference of dissociated GCs with nuclear factor kappaB (NF-kappaB)-driven gene activation. We show that dissociated GCs repress tumor necrosis factor-induced interleukin-6 gene expression by an NF-kappaB-dependent mechanism, without changing the expression level of inhibitor kappaB. The DNA-binding activity of induced NF-kappaB also remained unchanged after stimulation of cells with the various compounds. Evidence for a direct nuclear mechanism of action was obtained by analysis of cell lines constitutively expressing a fusion protein between the DNA-binding domain of the yeast Gal4 protein and the transactivating p65 subunit of NF-kappaB, which was able to efficiently repress a Gal4-dependent luciferase reporter gene upon addition of the dissociated compounds. We therefore conclude that, in addition to dissociating transactivation from activator protein 1 transrepression, dissociated GCs mediate inhibition of NF-kappaB signaling by a mechanism that is independent of inhibitor kappaB induction.

Animals↗
PubMed2000

Automated oxyhemoglobin dissociation curve construction to assess sickle cell anemia therapy.

Oxyhemoglobin dissociation curves measure the most important function of red blood cells - the affinity for oxygen and its delivery to the tissues. This function may be deranged in sickle cell anemia and some other hemoglobinopathies. An automated oxyhemoglobin dissociation curve analyzer constructed dissociation curves in 55 patients with hemoglobinopathies and in 24 control subjects while maintaining constant temperature and pH. Sigmoid curves were converted to rectilinear ones using the Hill equation. Oxygen affinity of red cells was assessed by calculation of P50 (the PO2 at which hemoglobin is half saturated). Results revealed separation of oxyhemoglobin dissociation Hill plots according to phenotype but with wide variability. Mean oxygen affinity of fetal hemoglobin was greatest, whereas that of sickle hemoglobin was least. Other hemoglobins were intermediate. A positive correlation between decreased oxygen affinity and carboxyhemoglobin confirmed the decreased oxygen affinity of sickle hemoglobin and decreased oxygen affinity and increased diphosphoglycerate in red cells. Hill plots are less sensitive discriminators of oxygen affinity than traditional sigmoid dissociation curves and offer no particular advantage. Serial studies in a subset of three sickle cell anemia patients treated conservatively suggest automated oxyhemoglobin dissociation curves may be useful in assessment of effectiveness of newer therapies of sickle cell anemia after refinement of the method and studies of larger populations.

Adult↗
PubMed1975

[Atrio-ventricular dissociation].

Cardiac rhythm disorders called atrioventricular dissociation, especially the so-called dissociation with interference have been causing controversy among the cardiologists for a long time. The latter term implies an independent activity of the atria and ventricles, that is in no way connected with an orthograde atrioventricular block, and that develops in the presence of intermittent normal conductivity of sinus impulses. This disorder is always of a secondary nature and it may be due to the disorders in the formation of the primary rhythm, or conductivity of the impulse (passive form), or acceleration of the secondary rhythm (active form). The causes of such atrioventricular dissociation may lie in sinus bradycardia, sinoauricular or incomplete atrioventricular block, compensatory pauses after extra-systoles, atrioventricular and ventricular tachycardia (paroxysmal or nonparoxysmal) with a retrograde atrioventricular block, etc. Examples of ECG patterns illustrating most of the above mechanisms are presented. Due to the existence of several mechanisms that cause atrioventricular dissociation, and due to the fact that individual authors imply different meanings by the term "interference", it seems reasonable to abandon the term "dissociation with interference". In such case it would seem more appropriate to call the state a primary rhythm or conductivity disorder indicating an incomplete atrioventricular dissociation for a proper choice of therapy.

Adult↗
PubMed1979

Dissociation of adult mammalian heart into single cell suspension: an ultrastructural study.

Adult rat heart was dissociated into a single cell suspension by a perfusion technique which used 0.05% collagenase and 0.1% hyaluronidase in Krebs-Ringer phosphate buffer (KRP). The non-muscle cells of the suspension were separated from the myocytes by centrifugation through 3% Ficoll solution in KRP with 0.01 mM Ca2+. An approximately 90% pure suspension of isolated single muscle cells was obtained with this method. The effects of the successive steps in the dissociation procedure on the ultrastructure of the heart were studied by scanning and transmission electron microscopy. After 30 minutes of enzyme digestion, dissociation of the inner endothelial lining of the ventricle into single cells or small groups of cells became apparent. In addition, the underlying cardiac skeleton began to disintegrate and linear arrays of cardiac muscle cells were observed. After 45 minutes of enzyme digestion the number of released single cells was higher because of the separation of intercalated discs. The majority of non-muscle cells were by now dissociated from the surfaces of muscle cells. Widening of the lateral intercellular spaces between the myocardial cells was associated with separation of desmosomes. In some regions of the heart, intact desmosomes, fasciae adherentes and gap junctions were observed even though lateral intercellular spaces had widened greatly. The majority of myocardial cells had become separated from one another after 60 minutes of enzyme digestion. Separation of gap junctional sites took place in two ways: (1) by 'unzipping' them through enzyme action; (2) by tearing them mechanically. Gap junction remnants were sometimes observed in a vesiculated state within the cell. The dissociation of the heart was ineffective when perfused with media containing 1.0 or 2 mM Ca2+. Alcian blue treatment after 60 minutes of enzyme digestion revealed that the basement membrane, and its accompanying collagen fibrils, was still present on the plasma membrane of dissociated single cells. The isolated myocardial cells retained their normal morphological characteristics. This study has enabled us to understand in detail how dismantlement of highly ordered adult cardiac tissue into a single cell suspension takes place. Cell suspensions of this type should be invaluable in the study of metabolic and synthetic activities in adult myocardial cells.

Animals↗
PubMed2006

[Effect of aluminum trichloride on dissociated Ca2+ in Hippocampus neuron cell as well as learning and memory].

OBJECTIVE: To investigate the effect of aluminum chloride on dissociated Ca(2+) in hippocampus neuron cells in mice and the relationship to the learning and memory. METHODS: Male ICR mice in the three intoxicated groups were administered with the double distilled water containing AlCl(3) (10, 50, 300 mg.kg(-1).d(-1)) while those in the control group were administered with the double distilled water for 100 days. The methods of behavior toxicology such as Morris swim maze were used for studying the effect of aluminum chloride on the changes of learning and memory in mice. With calcium sensitive fluorescence indicator Fura-2 as the fluorescent probe, the influence of the subchronic exposure to Al on the dissociated Ca(2+) in hippocampus neuron cells was observed. RESULTS: The dissociated Ca(2+) in hippocampus neuron cells in the middle dosage group and the high dosage group [(412.25 +/- 53.20), (467.37 +/- 32.85) times] was lower than those in the control group [(293.91 +/- 32.21) times] respectively (P < 0.01), and correlated positively with the dose and dissociated Ca(2+) (r = 0.861, P < 0.01). Compared with the control group, the latent period was lengthened (P < 0.05) in the middle dosage and the high dosage group. CONCLUSION: The subchronic exposure to AlCl(3) in mice affects the dissociated Ca(2+) in hippocampus neuron cells. The increase of dissociated Ca(2+) in hippocampus neuron cells may be correlated with the disfunction of cognition in the aluminium intoxicated mice.

Aluminum Chloride↗
PubMed2006

Improved circadian sleep-wake cycle in infants fed a day/night dissociated formula milk.

On the basis of the circadian nutritional variations present in breast milk, and of the implications for the sleep/wake cycle of the nutrients present in infant formula milks, we designed a formula milk nutritionally dissociated into a Day/Night composition. The goal was to improve the bottle-fed infant's sleep/wake circadian rhythm. A total of 21 infants aged 4-20 weeks with sleeping difficulties were enrolled in the three-week duration study. The sleep analysis was performed using an actimeter (Actiwatch) placed on an ankle of each infant to uninterruptedly record movements during the three weeks. The dissociated Day milk, designed to be administered from 06:00 to 18:00, contained low levels of tryptophan (1.5g/100g protein) and carbohydrates, high levels of proteins, and the nucleotides Cytidine 5 monophosphate, Guanosine 5 monophosphate and Inosine 5 monophosphate. The dissociated Night milk, designed to be administered from 18.00 to 06.00, contained high levels of tryptophan (3.4g/100g protein) and carbohydrates, low levels of protein, and the nucleotides Adenosine 5 monophosphate and Uridine 5 monophosphate. Three different milk-feeding experiments were performed in a double-blind procedure covering three weeks. In week 1 (control), the infants received both by day and by night a standard formula milk; in week 2 (inverse control), they received the dissociated milk inversely (Night/Day instead of Day/Night); and in week 3, they received the Day/Night dissociated formula concordant with the formula design. When the infants were receiving the Day/Night dissociated milk in concordance with their environment, they showed improvement in all the nocturnal sleep parameters analyzed: total hours of sleep, sleep efficiency, minutes of nocturnal immobility, nocturnal awakenings, and sleep latency. In conclusion, the use of a chronobiologically adjusted infant formula milk seems to be effective in improving the consolidation of the circadian sleep/wake cycle in bottle-fed infants.

Bottle Feeding↗
PubMedDate not supplied

Dissociation of neurological deficits in spinal decompression illness.

Functional differentiation is found in the spinal cord. A unique set of neurological deficits follows a multi-focal injury. Clinically, sensory and motor disturbance present independently, often resulting in sensory and motor deficit dissociation. This study examined 103 spinal decompression illness (DCI) cases. The neurological deficit dissociation was classified as follows: 1) Cases with sensory impairment only, or motor dysfunction alone, were tagged as having "dissociation" (44 cases); when a case was with both sensory and motor dysfunction, the spinal level of the sensory impairment was determined and was matched with the spinal segments responsible for the motor dysfunction; 2) If the two spinal areas did not match (i.e. with no regional overlap), they were tagged as having "dissociation" for each motor dysfunction (32 cases). In total, dissociation was present in 76 out of 103 cases. We concluded that clinical neurological deficit dissociation is frequently observed in spinal DCI.

Adolescent↗
PubMed1992

Cooperative cyanide dissociation from ferrous hemoglobin.

Rapid reduction of cyano-met hemoglobin (Hb+CN-) leads to the formation of an intermediate species, the cyanide derivative of ferrous hemoglobin, which dissociates to unliganded hemoglobin because of the extremely low affinity of the ligand for the ferrous heme iron. The properties of the intermediate were studied by transient spectroscopy in human hemoglobin and its isolated alpha and beta chains, in the presence and absence of CO. When mixing with dithionite, the time courses of reduction of the heme iron and dissociation of cyanide overlap considerably; addition to the reaction mixture of the redox indicator methyl viologen considerably increases the rate of reduction and allows unequivocal determination of the spectroscopic and kinetic properties of the intermediate. The results show that (i) the dissociation of cyanide from the isolated alpha and beta chains (as well as the (alpha CO)2(beta + CN-)2 hybrid) is a simple process; (ii) the two chains display similar rate parameters, but show spectroscopic inequivalence, both in the Soret and the visible regions; (iii) cooperative effects are shown to control the rate of dissociation of cyanide from hemoglobin, similarly to what happens for oxygen; and (iv) allosteric effectors (typically inositol hexaphosphate) increase the overall rate of dissociation by stabilization of the T state. We have, therefore, shown for the first time that the dissociation of cyanide from ferrous hemoglobin is controlled by the quaternary state, thereby adding one more ligand to the analysis of the structure-function relationships in hemoglobin.

Cyanides↗
PubMed1990

Guanine nucleotide regulation of B2 kinin receptors. Time-dependent formation of a guanine nucleotide-sensitive receptor state from which [3H]bradykinin dissociates slowly.

We have examined the binding of [3H]bradykinin to bovine myometrial membranes and assessed its sensitivity to guanine nucleotides. Total binding displayed a typical B2 kinin receptor specificity. However, saturation binding isotherms were resolved into at least two components with KD values of 8 pM (45%) and 378 pM (55%). Low affinity binding exhibited relatively rapid rates of association (kobs = 1.40 x 10(-2) s-1) and dissociation (k-1 = 3.82 x 10(-3) s-1), while high affinity binding exhibited considerably slower rates (kobs = 9.52 x 10(-4) s-1 and k-1 = 4.43 x 10(-5) s-1). Pre-equilibrium dissociation kinetics revealed that formation of high affinity binding was characterized as a time-dependent accumulation of the slow dissociation rate at the expense of at least one other more rapid dissociation rate. In the presence of 10 microM guanyl-5'-yl imidodiphosphate (Gpp(NH)p), at least two binding components were resolved with KD values of 37 pM (12%) and 444 pM (88%). Gpp(NH)p apparently specifically perturbed high affinity binding by completely preventing the accumulation of the slow dissociation phase. Instead, two more rapid dissociation rates (k-1 = 8.53 x 10(-3) s-1 and 4.43 x 10(-4) s-1) were observed. These results suggest that [3H]bradykinin interacts with at least two B2 kinin receptor-like binding sites in bovine myometrial membranes. A three-state model for the guanine nucleotide-sensitive agonist interaction with the high affinity binding sites is proposed.

Adenosine Triphosphate↗
PubMed1989

Receptor binding of [3H]naloxone benzoylhydrazone: a reversible kappa and slowly dissociable mu opiate.

In standard 3H-opioid binding assays, the benzoylhydrazone derivative of naloxone (6-desoxy-6-benzoylhydrazido-N-allyl-14-hydroxydihydronormorphi none; NalBzoH) inhibited mu, kappa, and delta binding at nanomolar concentrations. At concentrations as low as 1 nM, it also produced a wash-resistant inhibition of opioid binding. [3H]NalBzoH binding typically gave a ratio of total to nonspecific binding of 8:1. Binding reached steady state levels by 1 hr and was linear with tissue concentration. [3H]NalBzoH labeled two classes of sites. The binding to one was easily reversible whereas the other was not and was termed pseudoirreversible. At 25 degrees, almost 90% of [3H]naloxone binding and approximately 60-75% of [3H]NalBzoH binding dissociated over 90 min. However, the remainder of [3H]NalBzoH binding, corresponding to pseudoirreversible binding, remained constant over the next 5 hr at 25 degrees and additional studies suggested a dissociation half-life of approximately 24 hr. Competition studies indicated that the reversible binding corresponded to neither mu nor delta binding and may represent a novel subtype of kappa receptor. Pseudoirreversible binding was predominantly to a combination of both mu 1 and mu 2 receptors. Despite its extremely slow rate of dissociation, pseudoirreversible binding was not covalent inasmuch as lowering the pH to 5 or adding the GTP analog 5'-guanylylimidodiphosphate [Gpp(NH)p] completely dissociated prebound [3H] NalBzoH. The ability of Gpp(NH)p to dissociate pseudoirreversible [3H]NalBzoH binding raised the possibility that the slow rate of dissociation was related to interactions with a guanine nucleotide-binding protein.

3,4-Dichloro-N-methyl-N-(2-(1-pyrrolidinyl)-cycloh↗
PubMed1987

Dissociation of phosphate from beef heart mitochondrial F1-ATPase. Effect of adenine nucleotides.

The effect of exposure to adenine nucleotides on the dissociation of phosphate bound to beef heart mitochondrial F1-ATPase (F1) has been studied using a layered Sephadex centrifuge column method. The order of effectiveness of nucleotides in facilitating Pi dissociation from F1 was ATP greater than AMP-PNP much greater than ADP (where AMP-PNP indicates adenosine 5'-(beta,gamma-imido)triphosphate) when phosphate had bound to F1 as Pi from the medium. When ATP was present in amounts substoichiometric to F1, essentially all of the ATP present bound to F1, and the molar ratio of Pi released to ATP bound was near one. Under similar conditions AMP-PNP bound equally well to F1, but in order to effect similar amounts of Pi release it was necessary to use approximately 10 times as much AMP-PNP as ATP. The order of effectiveness of nucleotides in facilitating dissociation of phosphate which had bound to F1 as the gamma-phosphate of ATP was ATP = AMP-PNP much greater than ADP. In this case ATP and AMP-PNP were as effective in facilitating phosphate dissociation as was ATP in the case in which phosphate had bound to F1 as Pi. It is concluded that when phosphate has bound to F1 as Pi, binding of one molecule of ATP or more than one molecule of AMP-PNP are sufficient to facilitate dissociation of phosphate. When phosphate has bound to F1 as the gamma-phosphate of ATP, binding of one molecule of either ATP or AMP-PNP is sufficient to facilitate dissociation of phosphate.

Adenine Nucleotides↗
PubMed1986

Anti-hapten IgG antibodies bound to cell surface hapten: anti-IgG antibody prevents dissociation as measured with fluid phase hapten.

We investigated the dissociation by fluid phase hapten of IgG antibodies bound to cell surface hapten in the presence and absence of anti-IgG antibodies. Dissociation was quantitated with fluid phase hapten, preventing reassociation of the anti-hapten antibodies. More than 90% of the anti-hapten IgG alone was prevented from reassociation by low concentrations of fluid phase hapten (nanogram to microgram range). In contrast, no dissociation of some IgG-anti-IgG complexes could be measured even at 24 hr incubation in the presence of very large excess of fluid phase hapten (100 mg/ml). We excluded aggregate formation between anti-hapten antibodies due to cross-linking by anti-antibodies as a cause for decreased dissociability by 1) performing the experiments in large excess of anti-antibody, 2) showing that the phenomenon was independent of anti-hapten antibody density, 3) showing that decreased dissociation also occurred at 4 degrees C, and 4) showing that aggregation by protein A did induce decreased dissociability, albeit three orders of magnitude lower than the anti-antibody. It was concluded that anti-antibody directly affected the "avidity" of cell hapten bound anti-hapten IgG in an unknown manner.

Animals↗
PubMed1986

The nature of the antigen-antibody bond and the factors affecting its association and dissociation.

Considering that generally both Coulombic and van der Waals' bonds occur in AG-AB interactions, for AG-AB dissociation both interactions have to be made repulsive simultaneously. The theory and practice of repulsive van der Waals' interactions are outlined. Also treated are the hysteresis of AG-AB dissociation contrasted with the inhibition of association; dissociation of hydrogen bonds; dissociation by haptens; dissociation by alteration of tertiary and secondary configurations; and physical methods of dissociation.

Animals↗
PubMed1985

High and low affinity IL 2 receptors: analysis by IL 2 dissociation rate and reactivity with monoclonal anti-receptor antibody PC61.

In this report we characterize two classes of interleukin 2 (IL 2) binding sites on the basis of their differential IL 2 dissociation rate and of their reactivity with PC61, a monoclonal anti-IL 2 receptor (IL 2-R) antibody. PC61 inhibited the binding of IL 2 to both classes of receptor, but IL 2 did not inhibit the binding of PC61. This indicates that PC61 recognizes a determinant that is distal to the actual IL 2 binding site of the receptor. Dissociation experiments showed that the addition of excess unlabeled IL 2 resulted in a biphasic release of radiolabeled IL 2; 80 to 90% was dissociated rapidly (dissociation half-time, t1/2 of 60 sec) and the remainder more slowly (t1/2 60 to 90 min). The proportion of high and low affinity IL 2-R, as well as the relative difference in dissociation rates fit very well with the estimates derived previously from Scatchard plot analysis of equilibrium IL 2 binding. The addition of PC61 caused an accelerated dissociation of IL 2 from both high and low affinity IL 2-R (t1/2 of 16 and 120 sec respectively).

Animals↗
PubMed1986

Rod photoreceptor cells dissociated from mature mice retinas.

Intact rod photoreceptors were dissociated from pronase-treated whole retinas of adult mice by repeated passage through a plastic pipette tip. Hemocytometer counts of the cell suspensions indicate that, during a series of ten dissociation steps, a total of about 1-2 million intact photoreceptor cells are dissociated from one adult mouse retina, with less than 5% contamination from Müller cells and neurons of the inner retina. Visual cells with rod outer segments (ROS) and synaptic terminals are released in each step, but they occur in the greatest number during the sixth to ninth steps; detached ROS are released most frequently in the early steps, and neurons of the inner retinal layers appear in the later steps of dissociation. Nuclei are found in each step. Cell intactness was estimated by Trypan blue and Erythrosin B exclusion and by microscopic analysis using differential interference optics or scanning electron microscopy. The cells bind lectins (concanavalin A, Ricinis communis, and wheat germ agglutinin but not peanut agglutinin), displaying surface topography like that observed in situ. The metabolic capacity of dissociated cells was assessed by measuring the utilization of 32P inorganic phosphate for the synthesis of phospholipids and for the light-dependent phosphorylation of rhodopsin. Mature photoreceptor cells were estimated to contain, on average, 6.4 X 10(-12) g DNA, 2.3 X 10(-12) g RNA and 42-64 X 10(-12) g protein. The dissociation procedure provides a population of photoreceptor cells that appears suitable for microscopic, electrophysiological, and biochemical analysis.

Animals↗
PubMed1982

Mechanisms for dissociating proteoglycan aggregates.

Preformed Cs2SO4 zonal gradients were used to purify aggregates from proteoglycan preparations derived from associative extracts of the Swarm rat chondrosarcoma. Zonal gradients were used in solvents with different concentrations of guanidine HCl and different solvent pH values to study the mechanisms for dissociating the aggregates. Aggregates are stable in concentrations of guanidine HCl up to 1.5 M at pH 6.6. At 2 M guanidine HCl, partial dissociation occurs over 20 h in which a link protein is completely dissociated for every monomer proteoglycan dissociated from the aggregate structure. This suggests that in this solvent disaggregation occurs concurrent with complete separation of link protein from monomer. At solvent pH 2.7 to 3.3 in ionic conditions which normally promote aggregation, dissociation occurs by a mechanism in which the link protein remains associated with monomer. Thus, link protein-monomer complexes dissociate as bimolecular units from hyaluronic acid; such complexes then exhibit physical properties indistinguishable from pure monomers. The link protein-monomer complexes reassociate with hyaluronic acid to form link-stabilized aggregates when the solvent pH is raised to pH 7, i.e. to associative conditions. The study provides additional evidence for the role that link protein-monomer interactions have in proteoglycan aggregate structures.

Animals↗
PubMed1980

Relationship between deactivation of insulin-stimulated glucose transport and insulin dissociation in isolated rat adipocytes.

UNLABELLED: The time course of 125I-insulin dissociation from receptors and deactivation of insulin-stimulated glucose transport was measured in rat adipocytes. When cells were incubated with a submaximally stimulating insulin concentration (1 ng/ml), insulin dissociated rapidly at 24 degrees C and 37 degrees C, with a t1/2 of 26 and 14 min, respectively. On the other hand, deactivation of 3-O-methylglucose transport proceeded at a much slower rate. The t1/2 of deactivation was 73 min at 24 degrees C and 43 min at 37 degrees C. Thus, the activated state of the glucose transport system persisted at a time when receptor occupancy had greatly decreased, and hormone dissociation was 3 times faster than deactivation; both processes were equally temperature-dependent. When glucose (1 mM) was omitted from the buffer, deactivation was essentially completely inhibited for at least 2 h despite the fact that the rate of insulin dissociation was unaffected. IN CONCLUSION: 1) termination of the insulin signal on glucose transport can be separated from dissociation of insulin receptor complexes; 2) transport deactivation proceeds much more slowly than insulin dissociation and is dependent on some aspect of ongoing cellular metabolism.

Adipose Tissue↗