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Synaptically released histamine increases dye coupling among vasopressinergic neurons of the supraoptic nucleus: mediation by H1 receptors and cyclic nucleotides.

Activating direct olfactory (glutamatergic) inputs to supraoptic nucleus (SON) neurons increases interneuronal coupling in slices from lactating but from not virgin or male rats. Studied here were influences on coupling of another monosynaptic input to SON, the histaminergic tuberomammillary nucleus (TM) projection, activation of which selectively excites phasically firing (putative vasopressin) cells. Effects of TM stimulation and its possible downstream consequences on Lucifer yellow (LY) dye coupling among putative vasopressin cells were determined in male rat SONs. In unstimulated slices, 12 LY injections (1 cell/SON) yielded eight single and four pairs of coupled neurons. In slices in which TM was stimulated for 10 min at 10 Hz, 13 injections yielded 4 single and 28 coupled cells, with groups of 2 to 4 cells coupled to the injected neuron, a threefold increase in the number of coupled cells per injection (p < 0.02). Bathing slices in medium containing 10 microM pyrilamine (H1 antagonist) blocked this stimulation-induced coupling increase, suggesting mediation by activation of guanylate cyclase-cGMP to which H1 receptors often are linked . Bathing slices in medium containing 0.5-1 mM 8-bromo-cGMP yielded results similar to those of TM stimulation, a 2.5-fold increase over control in the number of coupled cells per injection. Effects of TM stimulation on coupling also were blocked by bathing slices in a guanylate cyclase inhibitor (10 microM LY83583). In contrast to cGMP, 1 mM 8-bromo-cAMP significantly reduced coupling. We conclude that synaptically released histamine increases coupling via cGMP-dependent mechanisms.

Animals↗

Tracer coupling pattern of amacrine and ganglion cells in the rabbit retina.

We examined the tracer coupling pattern of more than 15 morphological types of amacrine and ganglion cells in the rabbit retina. Individual cells were injected intracellularly with the biotinylated tracer Neurobiotin, which was then allowed to diffuse across gap junctions to label neighboring neurons. We found that homologous and/or heterologous tracer coupling was common for most proximal neurons. In fact, the starburst amacrine cell was the only amacrine cell type that showed no evidence of coupling. The remaining types of amacrine cell were coupled exclusively to other amacrines, either homologously or, more often, through a combination of homologous and heterologous junctions. In only one case did we visualize labeled ganglion cells following injection of Neurobiotin into an amacrine cell. In contrast, injection of Neurobiotin into ganglion cells almost always resulted in the labeling of amacrine cells. Taken together, these results suggest a directionality to the movement of tracer across gap junctions connecting amacrine and ganglion cells. We found that the coupling pattern for a given morphological type of cell was generally stereotypic and consistent across retinas. The notable exceptions to this finding were alpha ganglion cells and cells with morphology corresponding to that of on-off direction selective ganglion cells. In both cases, individual cells showed either extensive coupling to both amacrine and ganglion cells or no coupling at all. A notable finding was that, in every case, the neighboring cells within a tracer-coupled array were always within one gap junction of the injected neuron. Furthermore, in many cases, the array formed by the somata of tracer-coupled cells was almost perfectly coincident with the dendritic arbor of the injected cell. Thus, our results indicate that whereas coupling is extensive within the proximal retina, individual cells partake in coupled networks that are stereotypic and highly circumscribed.

Animals↗

Personal experiences of cystic fibrosis (CF) carrier couples prospectively identified in CF families.

This qualitative study explores the experiences of cystic fibrosis (CF) carrier couples, prospectively identified in CF families, and the impact of the resulting genetic risk on reproductive behavior. Of the 12 couples identified until 1997, seven couples participated in semistructured interviews and two couples filled in a questionnaire, two to eight years after receipt of the test-results. After receiving the results, most couples reported that they were shocked, because they did not expect to both be carriers. More anxiety was expressed by those who were pregnant (n = 4) at the time of testing. There were reported difficulties in disclosing the results to family members, and the reactions of family members were not always supportive. After testing, some couples had problems with reproductive decision-making. All viable pregnancies (17 in 8 couples) were monitored by prenatal diagnosis; all affected pregnancies were terminated (6 in 4 couples). Couples who have live-born children after testing may subsequently have concerns during infancy about the correctness of the results of prenatal diagnosis and how to inform their children. Most couples did not regret the testing and, in general, the counseling was experienced positively, although some dissatisfaction was reported with regard to the psychological support received during pregnancy. Couples supported the idea of carrier screening in the general population, although various concerns were expressed. The results indicate a preference for testing before pregnancy. These findings may be useful in investigating possible dilemmas caused by the introduction of population carrier screening. Observations reported here might also apply to other recessively inherited disorders.

Cystic Fibrosis↗

Couples' experiences of predictive testing and living with the risk or reality of Huntington disease: a qualitative study.

A qualitative study, based on family systems theory, was undertaken in order to gain a better understanding of the impact of predictive testing and of living with the risk or reality of Huntington disease (HD), on couple relationships. Semi-structured interviews were conducted with 14 couples; in 9 couples the at-risk partner had undergone testing, and of these, 4 were already affected with HD. At-risk partners in the remaining five couples had not been tested. Interview transcripts were analyzed to obtain a range of themes, which reflect the salient experiences of these couples in relation to HD. Most couples reported that receiving a predictive test result had little or no adverse effect on their relationship. However for two couples who separated after the at-risk partner received a non-carrier result, emotional factors associated with years of living with the HD risk, rather than the result itself, were regarded as having caused irreparable damage to the relationship. For two couples who have remained together since the diagnosis of one partner, loyalty was identified as the main factor contributing to the continuance of the relationship. The separations of the other two couples in which one partner was diagnosed were attributed to emotional distancing, and to the obsessive behavior of the affected partner. The findings of this study highlight both the individuality and the complexity of psychological effects on the intimate relationships of couples who live with the risk or reality of HD, and provide important insights for professionals offering support to these couples.

Adult↗

Morphology and tracer coupling pattern of alpha ganglion cells in the mouse retina.

Alpha cells are a type of ganglion cell whose morphology appears to be conserved across a number of mammalian retinas. In particular, alpha cells display the largest somata and dendritic arbors at a given eccentricity and tile the retina as independent on- (ON) and off-center (OFF) subtypes. Mammalian alpha cells also express a variable tracer coupling pattern, which often includes homologous (same cell type) coupling to a few neighboring alpha cells and extensive heterologous (different cell type) coupling to two to three amacrine cell types. Here, we use the gap junction-permeant tracer Neurobiotin to determine the architecture and coupling pattern of alpha cells in the mouse retina. We find that alpha cells show the same somatic and dendritic architecture described previously in the mammal. However, alpha cells show varied tracer coupling patterns related to their ON and OFF physiologies. ON alpha cells show no evidence of homologous tracer coupling but are coupled heterologously to at least two types of amacrine cell whose somata lie within the ganglion cell layer. In contrast, OFF alpha cells are coupled to one another in circumscribed arrays as well as to two to three types of amacrine cell with somata occupying the inner nuclear layer. We find that homologous coupling between OFF alpha cells is unaltered in the connexin36 (Cx36) knockout (KO) mouse retina, indicating that it is not dependent on Cx36. However, a subset of the heterologous coupling of ON alpha cells and all the heterologous coupling of OFF alpha cells are eliminated in the KO retina, suggesting that Cx36 comprises most of the junctions made with amacrine cells.

Amacrine Cells↗

Electrochemical properties of suprastructures galvanically coupled to a titanium implant.

In recent years, dental implants have been widely used for the aesthetic and functional restoration of edentulous patients. Dental implants and restorative alloys are required with high corrosion resistance. Suprastructures and implants of different compositions in electrical contact may develop galvanic or coupled corrosion problems. In addition to galvanic corrosion, crevice and pitting corrosion may occur in the marginal gap between dental implant assemblies. In this study, gold, silver-palladium, cobalt-chromium, and nickel-chromium suprastructures were used to investigate their galvanic and crevice corrosion characteristics in combination with titanium (Ti) implants. Potentiodynamic and potentiostatic testing were performed in artificial saliva at 37 degrees C. Potentiodynamic testing was carried out at the potential scan rate of 1 mV/s in the range of -600-1600 mV (SCE). Potentiostatic testing was performed with an open-circuit potential and current densities at -250, 0, and 250 mV (SCE) in artificial saliva. After electrochemical testing, surface morphologies and cross-sections were examined using micrographs of the samples. Potentiodynamic test results indicated that suprastructure/Ti implant couples produced passive current densities in the range of 0.5-12 microA/cm(2); Ti abutment/Ti implant and gold/Ti implant couples exhibited relatively low passive current densities; Co-Cr/Ti implant couples the highest. Co-Cr and Ni-Cr/Ti implant couples showed breakdown potentials of 700 and 570 mV (SCE), respectively. The open-circuit potentials of silver, Ti abutment, gold, Ni-Cr, and Co-Cr/Ti implant couples were -93.2 +/- 93.9, -123.7 +/- 58.8, -140.0 +/- 80.6, -223.5 +/- 35.1, and -312.7 +/- 29.8 mV (SCE), respectively, and did not change with immersion time. The couples exhibited cathodic current densities at -250 mV (SCE); in particular, gold and silver alloys showed high cathodic current densities of -3.18 and -6.63 microA/cm(2), respectively. At 250 mV (SCE), Ti abutment/Ti implant couples exhibited a minimum current density of 9.48 x 10(-2) microA/cm(2), but gold, Ni-Cr, Co-Cr, and silver/Ti implant couples exhibited 0.313, 1.27, 5.60, and 8.06 microA/cm(2), respectively. All couples exhibited relatively low current densities at 0 mV (SCE). Photomicrographs after electrochemical testing showed crevice or pitting corrosion in the marginal gap and at the suprastructure surface. Although of the tested samples Co-Cr/Ti implant couples showed the possibility of galvanic corrosion, its degree was not significant. However, it should be borne in mind that galvanic corrosion can accelerate localized corrosion, such as crevice or pitting corrosion.

Alloys↗

Changing patterns of ganglion cell coupling and connexin expression during chick retinal development.

We have used dye injection and immunolabeling to investigate the relationship between connexin (Cx) expression and dye coupling between ganglion cells (GCs) and other cells of the embryonic chick retina between embryonic days 5 and 14 (E5-14). At E5, GCs were usually coupled, via soma-somatic or dendro-somatic contacts, to only one or two other cells. Coupling increased with time until E11 when GCs were often coupled to more than a dozen other cells with somata in the ganglion cell layer (GCL) or inner nuclear layer (INL). These coupled clusters occupied large areas of the retina and coupling was via dendro-dendritic contacts. By E14, after the onset of synaptogenesis and at a time of marked cell death, dye coupling was markedly decreased with GCs coupled to three or four partners. At this time, coupling was usually to cells of the same morphology, whereas earlier coupling was heterogeneous. Between E5 and E11, GCs were sometimes coupled to cells of neuroepithelial morphology that spanned the thickness of the retina. The expression of Cx 26, 32, and 43 differed and their distribution changed during the period studied, showing correlation with events such as proliferation, migration, and synaptogenesis. These results suggest specific roles for gap junctions and Cx's during retinal development.

Animals↗

Mechanism of simultaneous iodination and coupling catalyzed by thyroid peroxidase.

Thyroid peroxidase (TPO) simultaneously catalyzes two very different types of reaction in the thyroid gland- iodination and coupling. The present study addresses the mechanism of this simultaneous dual activity. Compound I, the two-electron oxidation product of TPO, exists in two different forms--an oxoferryl porphyrin pi-cation radical and an oxoferryl protein radical. It has been proposed that iodination is mediated by the porphyrin pi-cation radical form of TPO compound I, while coupling is mediated by the protein radical form. However, results obtained in the present study favor the view that both iodination and coupling are mediated by the porphyrin pi-cation radical form of compound I. In the first part of the study, we compared coupling and iodination activities of two peroxidases with very similar crystal structures--cytochrome c peroxidase (CcP) and lignin peroxidase (LiP). Although these two peroxidases have very similar three-dimensional structures, CcP forms a compound I only of the protein radical type, whereas compound I of LiP exists only as a porphyrin pi-cation radical. Comparison of the catalytic activities of the two enzymes showed that diiodotyrosine (DIT)-stimulated coupling activity of LiP was significantly greater than that of CcP. Moreover, lignin peroxidase displayed very significant iodinating activity at acid pHs, whereas iodination with CcP was negligible at all pHs tested. Our findings with these two structurally similar peroxidases suggested that TPO-catalyzed iodination and coupling could both be mediated by the porphyrin pi-cation radical form of compound I. More direct evidence in support of this view was obtained in the second part of this study, employing TPO and lactoperoxidase (LPO) model systems in which iodination and coupling occurred simultaneously. Heme spectral analysis was used to correlate formation of the protein radical form of compound I with the kinetics of the iodination and coupling reactions. Formation of the compound I protein radical was not observed until the iodination and coupling reactions had almost been completed. In separate experiments it was shown that the spontaneous conversion of the porphyrin pi-cation radical form of TPO or LPO compound I to the protein radical form was markedly inhibited by a low concentration of iodide, especially in the presence of an iodide acceptor. These studies provide compelling evidence that both iodination and coupling are mediated by the porphyrin pi-cation radical form of compound I. This was further substantiated by the finding that coupling was inhibited in the presence of excess iodide, an observation readily explained by competition between iodide and DIT residues in thyroglobulin for oxidation by the porphyrin pi-cation radical.

Animals↗

Amine modulation of electrical coupling in the pyloric network of the lobster stomatogastric ganglion.

1. The neurons of the pyloric network of the lobster (Panulirus interruptus) stomatogastric ganglion organize their rhythmic motor output using both chemical and electrical synapses. The 6 electrical synapses within this network help set the firing phases of the pyloric neurons during each rhythmic cycle. We examined the modulatory effects of the amines dopamine (DA), serotonin (5HT) and octopamine (Oct) on coupling at all the electrical synapses of the pyloric network. 2. Electrical coupling within the pacemaker group [anterior burster (AB) to pyloric dilator (PD), and PD-PD] was non-rectifying, while coupling at the other electrical synapses [AB to ventral dilator (VD), PD-VD, lateral pyloric (LP) to pyloric (PY), and PY-PY] was rectifying. 3. Dopamine decreased or increased the coupling strength of all the pyloric electrical synapses: the sign of the effect depended upon which neuron was the target of current injection. For example, DA decreased AB-->PD coupling (i.e., when current was injected into the AB) but increased coupling in the other direction, PD-->AB. Dopamine decreased AB to VD coupling when current was injected into either neuron. Serotonin also had mixed effects; it enhanced PD-->AB coupling but decreased AB to VD and PD to VD coupling in both directions. Octopamine's only effect was to reduce PD-->VD coupling. 4. Dopamine increased the input resistance of the AB neuron but decreased the input resistance of the PD and VD neurons. Serotonin reduced the input resistance of the VD and PY neurons, while Oct did not significantly change the input resistance of any pyloric neuron. 5. The characteristic modulation of electrical coupling by each amine may contribute to the unique motor pattern that DA, 5HT and Oct each elicit from the pyloric motor network.

Amines↗

Cardioventilatory coupling in the anaesthetised rabbit, rat and guinea-pig.

Cardioventilatory coupling is a temporal coherence of respiratory and cardiac rhythms, seen in humans at rest, and during sleep and anaesthesia. In this study we compared the cardioventilatory coupling of anaesthetised rabbits, rats and guinea-pigs. Breathing two successive anaesthetic concentrations (1 or 2% isoflurane) we compared the effect of anaesthetic depth and species on (1) heart rate, (2) heart rate variability, (3) ventilatory rate (f), (4) ventilatory variability, (5) ratio HR/f, (6) degree of coupling (Shannon entropy of the distribution of intervals between inspiration and the preceding electrocardiographic R wave - the RI interval) and (7) coupling pattern, classified into four sub-patterns (I-IV) based upon inspection of the RI interval time series. Rabbits exhibited significantly less ventilatory variability and coupling than rats or guinea-pigs. The sub-pattern of coupling also differed between the three species. Rabbits showed coupling only when HR and f were close to integer ratios whereas other species coupled at non-integer ratios. Ventilatory variability in the rat and guinea-pig differed according to the pattern of coupling observed. Of the three species studied, the rat and guinea-pig demonstrated coupling most similar to that of anaesthetised human subjects. Anaesthetic concentration did not influence the pattern or degree of coupling.

Anesthesia↗

Anatomical and electrotonic coupling in developing genioglossal motoneurons of the rat.

Dye-, tracer- and electrotonic coupling were studied independently in genioglossal (GG) motoneurons using intracellular recordings in in vitro brainstem slices from rats postnatal ages 1-30 days. The subpopulation of GG motoneurons were retrogradely labeled after an injection of dextran-rhodamine into the posterior tongue. Dye-coupling was studied with Lucifer yellow injected into 55 motoneurons and tracer-coupling with neurobiotin injected into 89 presumptive GG motoneurons. Of the motoneurons injected with Lucifer yellow, only 6 of 41 cells (16.2%) exhibited dye-coupling; all occurred in animals less than 9 days old. In all but one instance, dye-coupling was restricted to only one other cell. No evidence of dye-coupling was found in the 14 cells injected in animals older than 8 days. Tracer-coupling (neurobiotin) was demonstrated in 12 of 30 cells (40%) from animals 1-2 days old and in 6 of 21 cells (28.6%) from animals 3-8 days old. Of the remaining 38 cells from animals 10 days of age and older, only one cell was found to be tracer-coupled. Cells injected with neurobiotin were coupled to an average of two other cells. Electrotonic coupling, as demonstrated with a short latency depolarization (SLD) in response to stimulation of hypoglossal axons, was found in developing GG motoneurons. These SLDs were revealed in 17 of 40 GG motoneurons (42.5%) examined in 1-8-day-old animals. There were no SLDs recorded in the 10 cells examined from animals of 10 days and older. The significance of coupling relative to patency of the newborn upper airways is discussed.

Animals↗

Psychological aspects of donor insemination: evaluation and follow-up of recipient couples.

OBJECTIVE: To evaluate the utility of psychological screening for couples entering a donor insemination program. DESIGN: Each spouse completed questionnaires. A psychologist reviewed them and rated the psychological fitness of the couple for participation in the program. Follow-up questionnaires were sent to each couple at a mean of 11 months after entry into the program. SETTING: Applicants for donor insemination were studied in an infertility program in a large, tertiary referral center. PATIENTS, PARTICIPANTS: Consecutive applicants to enter the donor insemination program were required to participate in the initial evaluation. INTERVENTIONS: Couples judged by the psychologist to be at risk for a poor psychological outcome had an assessment and counseling interview with the psychologist before proceeding with insemination. MAIN OUTCOME MEASURES: Initially, the Stress and Infertility Questionnaire measured specific anxieties related to donor insemination, marital and sexual impact, and attitudes about confidentiality; the Brief Symptom Inventory measured psychological distress; and the Dyadic Adjustment Inventory assessed marital satisfaction. At follow-up, 48% of couples returned a modified version of the Stress and Infertility Questionnaire and the other two questionnaires. RESULTS: The psychologist's rating was predictive of pregnancy rates (59% for excellent candidates, 41% for acceptable couples, and 14% for couples psychologically at risk). At-risk couples were more likely to drop out of the program (50% versus only 20% of other couples). Sexual problems were reported by 59% of women and 53% of men. Couples believed that a child should not be told of the donor insemination (74% of wives and 80% of husbands). Initially, 64% of wives and 70% of husbands chose total secrecy with families or friends, and these attitudes shifted little over time. CONCLUSION: This screening procedure is cost-effective and suggests that psychological intervention should be attempted with at-risk couples.

Adult↗

Voluntary counseling and testing for couples: a high-leverage intervention for HIV/AIDS prevention in sub-Saharan Africa.

Most HIV infections in sub-Saharan Africa occur during heterosexual intercourse between persons in couple relationships. Women who are infected by HIV seropositive partners risk infecting their infants in turn. Despite their salience as social contexts for sexual activity and HIV infection, couple relationships have not been given adequate attention by social/behavioral research in sub-Saharan Africa. Increasingly studies point to the value of voluntary HIV counseling and testing (VCT) as a HIV prevention tool. Studies in Africa frequently report that VCT is associated with reduced risk behaviors and lower rates of seroconversion among HIV serodiscordant couples. Many of these studies point out that VCT has considerable potential for HIV prevention among other heterosexual couples, and recommend that VCT for couples be practiced more widely in Africa. However, follow-up in the area of VCT for couples has been extremely limited. Thus, current understandings from social/behavioral research on how couples in sub-Saharan Africa manage HIV risks as well as HIV prevention interventions to support couples' HIV prevention efforts have remained underdeveloped. It appears that important opportunities are being missed for preventing HIV infection, be it by heterosexual transmission or mother-to-child HIV transmission by mothers who have been infected by their partners. Based on an overview of documentation on VCT in sub-Saharan Africa, this paper proposes that increased attention to couples-focused VCT provides a high-leverage HIV prevention intervention for African countries. The second half of the paper indicates areas where VCT needs to be strengthened, particularly with respect to couples. It also identifies areas where applied social/behavioral research is needed to improve knowledge about how couples in sub-Saharan Africa deal with the risks of HIV infection.

Acquired Immunodeficiency Syndrome↗

Evidence for coupled electron and proton transfer in the [8Fe-7S] cluster of nitrogenase.

Substrate reduction by nitrogenase requires electron transfer from a [4Fe-4S] cluster in the iron (Fe) protein component to an FeMo cofactor in the molybdenum-iron (MoFe) protein component in a reaction that is coupled to MgATP hydrolysis and component protein association and dissociation. An [8Fe-7S] (or P-) cluster in the MoFe protein has been proposed as an intermediate electron-transfer site, although how this cluster functions in electron-transfer remains unclear. In the present work, it is demonstrated that one redox couple of the P-cluster (P2+/1+) undergoes coupled electron and proton transfer, whereas a more reduced couple (P1+/N) does not involve a coupled proton transfer. Redox titrations of the MoFe protein P-cluster were performed, and the midpoint potential of the P2+/1+ couple (Em2) was found to be pH dependent, ranging from -224 mV at pH 6.0 to -348 mV at pH 8.5. A plot of Em2 versus the pH for this redox couple was linear and revealed a change of -53 mV/pH unit, indicating a single protonation event associated with reduction. From this plot, it was concluded that p is <6.0 and p is >8.5 in a proton-modified Nernst equation. In contrast, the midpoint potential for the P1+/N couple (Em1) was found to be -290 mV and was invariant over the pH range 6.0-8.5. These results indicate that the protonated species does not influence either the P1+ or the PN oxidation states. In addition, at physiological pH values, electron transfer is coupled to proton transfer for the P2+/1+ couple. The P-clusters are unique among [Fe-S] clusters in that they appear to be ligated to the protein through a serinate-gammaO ligand (betaSer188) and a peptide bond amide-N ligand (alphaCys88), in addition to cysteinate-S ligands. Elimination of the serinate ligand by replacement with a glycine was found to shift the Em values for both P-cluster couples by greater than +60 mV, however the pH dependence of Em2 was unchanged. These results rule out Ser188 as the protonated ligand responsible for the pH dependence of Em2. The implications of these results in understanding the nitrogenase electron-transfer mechanism are discussed.

Amino Acid Substitution↗

How much backbone motion in ubiquitin is required to account for dipolar coupling data measured in multiple alignment media as assessed by independent cross-validation?

The magnitude of backbone internal motions in the small protein ubiquitin that needs to be invoked to account for dipolar coupling data measured in multiple alignment media is investigated using an intuitively straightforward approach. This involves simultaneous refinement of the coordinates (against NOE, torsion angle, and dipolar coupling restraints) and optimization of the magnitudes and orientations of the alignment tensors by means of torsion angle simulated annealing and Cartesian space minimization. We show that N-H dipolar couplings in 11 different alignment media and N-C', H(N)-C', and C alpha-C' dipolar coupling in two alignment media can be accounted for, at approximately the level of uncertainty in the experimental data, by a single structure representation. Extension to a two-member ensemble representation which provides the simplest description of anisotropic motions in the form of a two-site jump model (in which the overall calculated dipolar couplings are the averages of the calculated dipolar couplings of the individual ensemble members), results in modest, but significant, improvements in dipolar coupling R-factors for both the working set of couplings used in the refinement and for the free cross-validated set of C alpha-H alpha dipolar couplings recorded in two alignment media. Extensions to larger ensemble sizes do not result in any R-factor improvement for the cross-validated C alpha-H alpha dipolar couplings. With a few notable exceptions, the amplitudes of the anisotropic motions are small, with S(2)(jump) order parameters > or =0.8. Moreover, the structural impact of those few residues that do exhibit larger amplitude motions (S(2)(jump) ranging from 0.3 to 0.8) is minimal and can readily be accommodated by very small backbone atomic rms shifts (<0.5 A) because of compensatory changes in phi and psi backbone torsion angles. In addition, evidence for correlated motions of N-H bond vectors is observed. For most practical applications, however, refinement of NMR structures against dipolar couplings using a single structure representation is adequate and will not adversely impact coordinate accuracy within the limits of the NMR method.

Kinetics↗

Tumor promoters cause a rapid and reversible inhibition of the formation and maintenance of electrical cell coupling in culture.

The effect of tumor promoters on electrical coupling between human FL cells was investigated with a microelectrode technique. When a low concentration (100 ng/ml) of 12-O-tetradecanoylphorbol 13-acetate (TPA) was added to culture medium, only 6% of the cells showed electrical coupling after 5 hr, whereas in control medium more than 90% of the cells were coupled. In the presence of TPA, cell coupling remained suppressed for at least another 19 hr. When TPA was washed out from the culture medium, the cells commenced electrical coupling: 90% of the cells were coupled within 4 hr of the removal of TPA, a rate very similar to that of nontreated control cells. Therefore, TPA-mediated inhibition of cell coupling is reversible. When TPA was added to a culture in which more than 90% of the cells had already established electrical coupling, the percentage of coupled cells decreased to 6% within 8 hr, indicating that TPA can also diminish already established cell coupling. Inhibition of cell coupling also was achieved with other mouse skin tumor promoters--phorbol 12,13-didecanoate, ingenol dibenzoate, and mezerein--whereas nonpromoting derivatives--phorbol and 4 alpha-phorbol 12,13-didecanoate-showed no effect. None of these compounds changed the membrane potential, membrane resistance, or growth rate of FL cells. Thus, it appears that TPA and structurally-related tumor promoters specifically disturb the formation or function, or both, of cell-cell junctions, without significantly affecting the general properties of the surface membrane or the growth of FL cells.

Cell Division↗

Coupled oscillations in food webs: balancing competition and mutualism in simple ecological models.

As in other oscillating systems, oscillations of consumer resource pairs in ecological systems may be coupled such that complex behavior results. The form of that coupling may determine the nature and extent of this behavior. Two biologically significant forms of coupling are here investigated: first, where consumers consume each other's resources (CR coupling, representing competition between the two consumers), and second, where the resources are in competition with one another (RR coupling, potentially representing indirect mutualism between the two consumers). Interestingly, CR coupling leads to in-phase synchrony of the oscillations, whereas RR coupling leads to antiphase synchrony. With either form of coupling, if the coupling remains weak, synchronous behavior is generated in the two systems. At strong levels of coupling, when the two forms act simultaneously, a balance between competition and mutualism is generated, which is manifest differently at different levels of resource coupling.

Animals↗

Effects of increased pericardial pressure on the coupling between the ventricles.

STUDY OBJECTIVE: The mechanical coupling between the ventricles occurs directly through the myocardium (ventricular-ventricular coupling) and indirectly through the pericardium (ventricular-pericardial-ventricular coupling). We postulated that the magnitude of ventricular-pericardial-ventricular coupling would increase at high pericardial pressures, while ventricular-ventricular coupling would be unaltered. DESIGN: Canine hearts were removed and placed in cold cardioplegic solution. Balloons were inserted into each ventricle and the left and right ventricular pressure (dP1, dPr) and volume (dV1, dVr) changes caused by increasing the pressure and volume of the other ventricle and by increasing pericardial pressure (dPp) were measured. EXPERIMENTAL MATERIALS: Hearts from 10 random source dogs, weight 12.5-18 kg, were used. MEASUREMENT AND MAIN RESULTS: At control pericardial pressure levels, the magnitude of the pericardial-ventricular interactions was greater than the ventricular-ventricular interactions: dP1/dPp was significantly greater than dP1/dPr, at 0.71 (SEM 0.04), n = 6, v 0.18 (0.03), p less than 0.01, and dV1/dPp was significantly greater than dV1/dPr, at -0.83 (0.09) v -0.24 (0.06), p less than 0.05. Raising the pericardial pressure increased the mechanical coupling between the ventricles: dP1/dPr approximately, dV1/dPr approximately, dPr/dP1 approximately, and dVr/dP1 approximately increased significantly (p less than 0.05) by 0.48 (0.03), 0.67 (0.13), 0.38 (0.05), and 0.61 (0.09) respectively. This increased coupling occurred through pericardial pressure changes. If pericardial pressure was maintained constant, the coupling between the ventricles was unaltered. This same pattern was observed in four in situ experiments. For these experiments, at the raised pericardial pressure levels, dP1/dPr increased, from 0.51 (0.03) to 0.79 (0.01), p less than 0.05, if pericardial pressure was allowed to vary, but was unaltered with a constant pericardial pressure, at 0.42 (0.03) v 0.44 (0.04), p greater than 0.5. CONCLUSIONS: Ventricular interdependence was increased with raised pericardial pressure and this increased coupling was due primarily to an increased ventricular-pericardial-ventricular coupling. This increased coupling may help to explain the paradoxical pulse observed in cardiac tamponade.

Animals↗