Response of frog and toad skin to norepinephrine

Maintenance of a hydrated integument is essential to the normal function of amphibian skin, and amphibians have developed mechanisms to minimize cutaneous dessication. The present work was conducted on skins of amphibians exhibiting a clear preference for either of two such mechanisms to study the influence of such mechanisms on the characteristics of epithelial transport. The response to norepinephrine (NE) was studied in isolated skins of a semiaquatic frog (Leptodactylus chaquensis), known to maintain indispensable skin moisture by secreting a superficial film of mucus via sympathetic stimulation of skin glands, and a terrestrial toad (Bufo arenarum), which replenishes a superficial film of fluid by drawing soil water upward by capillarity. In L. chaquensis skin, NE 5.0 x 10(-7) M, induced slow onset, sustained increases in short-circuit current (SCC) and transepithelial conductance, which were abolished by amiloride, a specific sodium transport inhibitor. At 1.2 x 10(-5) M, the response to NE exhibited a faster onset and a shorter time course. The SCC response also became insensitive to amiloride and could thus be induced by exposing the skin to NE in the presence of the inhibitor. The response was also greatly reduced in the absence of chloride, strongly suggesting a greater dependence on the glandular secretory response. In B. arenarum skin, the response to NE was far more sensitive to amiloride, regardless of the concentration of NE used. Induction of a response in the amiloride-blocked skin required a 10-fold higher concentration of NE, and the resulting effect was still considerably smaller than that observed in the skin of L. chaquensis after the same treatment. The number of mucous glands per unit area in B. arenarum skin was found to be around one-fifth of that observed in L. chaquensis, thus in part explaining the difference in the magnitude of the responses. The response of the skin of L. chaquensis to NE in the presence of sulfate was found to be consistent with the postulated involvement of frog skin glands in sulfate excretion. In contrast, this function was not evident in the skin of B. arenarum. The pattern of response of B. arenarum skin to all concentrations of NE tested closely resembles that seen after exposure to agents known to activate a cyclic AMP-dependent, high-permeability Cl pathway previously described by us in the skin of the toad. Our observations underscore the physiological differences existing in skins from different species, particularly regarding the relative importance of the glandular component of transport.     

Actions of tachykinins on the ion transport across the frog skin

The tachykinin-dependent stimulation of ion transport across frog skin was studied. Tachykinin stimulation was due to interaction with an NK1-like receptor as [Sar9-Met(O2)11]-Substance P (a very selective NK1 agonist) strongly stimulated SCC, whereas [beta-Ala8]-Neurokinin A 4-10 (a very selective NK2 agonist) did not. The rank order of tachykinin potency was: PG-KI > Uperolein > Hylambatin > Kassinin > Phyllomedusin > [Sar9-Met(O2)11]-Substance P > Ranatachykinin A > Physalaemin > Ranakinin > Substance P and Eledoisin > Neurokinin A. Neurokinin B, Scyliorhinin I, Urechistachykinin I and Urechistachykinin II had no effect. We conclude that the minimal structural requirements for stimulating SCC in the frog skin were the presence of: a) the C-terminal sequence Phe-X-Gly-Leu-Met-NH2; b) at least one Pro residue in the N-terminal sequence.     

NADPH-diaphorase-positive neurons and pathways in the brain of the frog Rana esculenta

We described the NADPH-diaphorase-containing neurons and fibres in the brain of the frog Rana esculenta. In the telencephalon stained cells occurred in the olfactory bulb, all subdivisions of the pallium, the diagonal band, the medial septum and the striatum. The olfactory glomeruli showed the most intense enzyme reaction. The neuropil of the accessory olfactory bulb was also heavily stained and this staining extended to the rostral diencephalon through the ventral lateral pallium. Fibre staining was less intense in the medial pallium and the medial septum. In the diencephalon, NADPH-diaphorase staining was concentrated in the middle third of this part of the brain. The stained cells were embedded in a dense network of thin, stained fibres and terminals in the lateral anterior and central thalamic nuclei. Faintly stained cells were present also in the posterior preoptic nucleus, anterior thalamic nucleus, nucleus of Bellonci, corpus geniculatum thalamicum and the suprachismatic nucleus. In the mesencephalon, heavily stained cells occurred in the nucleus profundus mesencephali, anterodorsal, anteroventral and especially in the posterodorsal tegmental nuclei. Neuronal staining was less intense in the optic tectum and the torus semicircularis. Thick, intensely stained fibres occupied the lateral part of the tegmentum and the 7th layer of the tectum. A loose network of thin fibres occupied the periventricular area and all tegmental nuclei. In the rhombencephalon, the reticular nuclei and the inferior raphe nucleus showed the most intense staining, while some cells in the nucleus of the solitary tract and the dorsal column nuclei were less intensely stained. Heavy staining of fibres was characteristic of the spinal trigeminal tract, the solitary tract and the reticulospinal pathway.     

Modulation of cardiac performance by amiloride and several selected derivatives of amiloride

Amiloride and its derivatives (benzamil, dichlorobenzamil, 5-(N,N-dimethyl)-amiloride, 5-(N-ethyl-N-isopropyl)-amiloride, (N,N-hexamethylene)- amiloride and 5-(N-methyl-N-isobutyl)-amiloride) are commonly used as selective blockers of Na+/Ca++ exchange or Na+/H+ exchange. Very little information is currently available regarding their effects on cardiac performance. It was observed that addition of amiloride or any of the selected derivatives to the coronary perfusate of the right ventricular wall produced a potent depressive effect on peak developed tension and the rates of tension generation and dissipation. The concentrations at which this occurred are those that are commonly used in ischemia or hypoxia studies. Significantly, the depressive action of the drugs increased with the perfusion duration and never achieved a stable level. An initial, transient positive inotropic effect was observed with some of the drugs. If the drug concentration and perfusion time was limited, the effects were reversible. All of the drugs except amiloride produced extra systoles. The drugs were capable of blocking Ca++ transients in isolated cardiomyocytes but had little effect on intracellular pH. The drugs lengthened the action potential duration and decreased the action potential amplitude and upstroke velocity. Their effects on cardiac performance may involve a complex inhibition of Ca++ influx and K+ efflux in addition to a stimulation of a nonselective cation current. It is concluded that amiloride and its analogs have striking effects on cardiac performance which may be unrelated to their capacity to inhibit Na+/Ca++ or Na+/H+ exchange. In summary, the use of these drugs is not normally recommended in cell or tissue perfusion experiments because of their nonselectivity. However, if the drug concentration and perfusion time is controlled carefully, interpretable data may be obtained in some cases.     

Influence of centrifugal pathways on forward masking of ventral cochlear nucleus neurons

When responses to one part of a sequence of auditory signals reduce the responses to a subsequent portion of the signal, "forward masking" results. Although forward masking occurs in the auditory nerve, that observed in the ventral cochlear nucleus (VCN) more closely resembles psychophysical forward masking. In contrast to the auditory nerve in which the amount of forward masking is proportional to the amount of excitation produced by the masker, most VCN neurons show a poor correlation between forward masking and excitation produced by the masker, indicating a more complex interaction between responses to adjacent signals. This study tested the hypothesis that one component of forward masking is produced by inputs from centrifugal neural connections to the VCN. The centrifugal pathways were interrupted with knife-cut lesions medial to the CN. Responses of single units obtained 60 minutes after the lesions were compared to those obtained before the lesions. In primarylike, sustained chopper and on units the lesions resulted in a reduction in forward masking and enhanced recovery. In contrast, lesions resulted in increased masking in primarylike-notch and low-intensity chopper units. The relationship between masker-elicited excitation and forward masking became more monotonic for transient choppers and on units, approaching that observed for auditory nerve fibers. These effects are probably the result of removal of both inhibitory and excitatory inputs, ultimately reflecting a balance of excitation and inhibition to each neural population in the VCN.     

Influence of dopamine on the pulmonary pressure and shunt volume after cardiac surgery (author's transl)

After the administration of Dopamine to 20 patients after cardiac surgery pulmonary shunt volume was determined with regard to the cardiac output and pulmonary pressure. It was shown that the infusion of Dopamine leads to decrease of the arterio-venous oxygen difference (AVDO2) as well as to the decrease of the PO2 in the arterial blood. On the contrary, in venous bood, PO2, Oxygen saturation, content and pulmonary pressure increased. The increase of the pulmonary right-to-left shunt observed (Qs Qt form 14.9 to 19.2%) is in proportion with the increase in cardiac output. Hence we do not suppose a specific shunting effect of Dopamine. Our findings suggest, that in the case of an already reduced blood oxygen level a further fall of the arterial oxygen saturation caused by a shunt must be avoided by additional enrichment of oxygen content of the inspired air.     

Long-term acceptance of skin and cardiac allografts after blocking CD40 and CD28 pathways

The receptor-ligand pairs CD28-B7 and CD40-gp39 are essential for the initiation and amplification of T-cell-dependent immune responses. CD28-B7 interactions provide 'second signals' necessary for optimal T-cell activation and IL-2 production, whereas CD40-gp39 signals co-stimulate B-cell, macrophage, endothelial cell and T-cell activation. Nonetheless, blockade of either of these pathways alone is not sufficient to permit engraftment of highly immunogenic allografts. Here we report that simultaneous but not independent blockade of the CD28 and CD40 pathways effectively aborts T-cell clonal expansion in vitro and in vivo, promotes long-term survival of fully allogeneic skin grafts, and inhibits the development of chronic vascular rejection of primarily vascularized cardiac allografts. The requirement for simultaneous blockade of these pathways for effective inhibition of alloimmunity indicates that, although they are interrelated, the CD28 and CD40 pathways are critical independent regulators of T-cell-dependent immune responses.     

Influence of infusion solution on the vascular permeability in rat skin

Numerous hypotheses have been considered to explain the fundamental mechanism(s) for the development of systolic dysfunction and heart failure in animals and humans with arterial hypertension. Besides contractile disturbances of cardiomyocytes and interstitial and perivascular fibrosis, cardiomyocyte loss is now being considered as one of the determinants of the maladaptive processes implicated in the transition from compensated to decompensated left ventricular hypertrophy. A number of experimental evidence suggest that exaggerated apoptosis may account for the loss of cardiomyocytes in the hypertensive left ventricle. Furthermore, some factors intrinsic and extrinsic to the cardiomyocyte emerge as potential candidates to trigger apoptosis. The elucidation of the possible interactions between these factors may be of major interest to prevent the progression to heart failure in patients with hypertensive heart disease.     

Influence of external pressure on transcutaneous oxygen tension and laser Doppler flowmetry on sacral skin

The effects of external pressure upon transcutaneous oxygen tension (tcpO2) and skin blood flow (SKBF) over the sacral area were studied in 16 healthy volunteers. Pressure was applied on the sacral area using a special device which included tcpO2 and laser-Doppler flowmetry probes. The mean values of tcpO2, SKBF and wave occurrence of the flow were analysed for 18 increasing pressure levels. A significant decrease of tcpO2 was observed from 40 mmHg (5.3 Kpa) of applied pressure, while we obtained a significant decrease of SKBF when the external pressure was at 20 mmHg (2.7 Kpa). Using a non-linear regression model, we have found a fourth degree polynomial to describe the relationship between tcpO2 and SKBF according to increasing pressure. SKBF slow wave occurrence decreased when external pressure was at 10 mmHg (1.3 Kpa), while rapid wave occurrence significantly decreased only at 120 mmHg (16.1 kPa) pressure.     

Influence of solubility and permeant size on absorption and metabolism of xenobiotics in rabbit skin

1 In order to obtain an understanding of the risks associated with dermal uptake of xenobiotics it is necessary to investigate the principles governing percutaneous absorption and metabolism. The single-pass perfused rabbit ear served as a convenient model for the simultaneous study of absorption and metabolism of a variety of substances. We found: 2 When substances are applied in a lipophilic vehicle the permeation coefficient Kr is notably decreased when the distribution coefficient (n-octanol/perfusion buffer (pH 7.4)) Poct. exceeds 2000, Kr also decreases with increasing molecular weight. 3 Substances which are in the ionized state at pH 7.4 may have a higher permeation coefficient than would be expected from Poct. or by molecular weight. 4 The partition of a drug from the stratum corneum to the epidermal enzymes controls the extent of its metabolism. However, substances with increased solubility at physiological pH are metabolized to a lesser extent than expected from their absorption rate. 5 The decrease in percutaneous absorption and metabolism of a substance with a Poct. exceeding 2000 is consistent with the assumption that high lipophilicity hinders partition of the substance from the stratum corneum to the viable metabolizing tissue.     

Effect of oxytocin on transepithelial transport of water and Na+ in distinct ventral regions of frog skin (Rana catesbeiana)

Thoracic, abdominal, and pelvic fragments of ventral skin of Rana catesbeiana were analysed regarding the effect of oxytocin on: (1) transepithelial water transport; (2) short-circuit current; (3) skin conductance and electrical potential difference; (4) Na+ conductance, the electromotive force of the Na+ transport mechanism, and shunt conductance; (5) short-circuit current responses to fast Na+ by K+ replacement in the outer compartment, and (6) epithelial microstructure. Unstimulated water and Na+ permeabilities were low along the ventral skin. Hydrosmotic and natriferic responses to oxytocin increased from thorax to pelvis. Unstimulated Na+ conductance was greater in pelvis than in abdomen, the other electrical parameters being essentially similar in both skin fragments. Contribution of shunt conductance to total skin conductance was higher in abdominal than in pelvic skin. Oxytocininduced increases of total skin conductance, Na+ conductance, and shunt conductance in pelvis were significantly larger than in abdomen. An oscillatory behaviour of the short-circuit current was observed only in oxytocin-treated pelvic skins. Decrease of epithelial thickness and increase of mitochondria-rich cell number were observed from thorax to pelvis. Oxytocin-induced increases of interspaces were more conspicuous in pelvis and abdomen than in thorax.     

Effects of electroconvulsive therapy on plasma vasopressin and oxytocin

BACKGROUND: Animal studies suggest that vasopressin has cognitive-enhancing properties and oxytocin may have amnestic effects. A clinical report suggests that the acute increase in oxytocin-associated neurophysin predicts clinical response to electroconvulsive therapy (ECT) in depressed patients. METHODS: Medication-free patients with major depression were randomized to receive right unilateral or bilateral ECT administered with electrical stimulus intensity at either just above seizure threshold or at 150% above seizure threshold. The associations between plasma vasopressin, oxytocin, ECT treatment parameters, clinical outcome, and cognitive effects were assessed. RESULTS: The sample comprised 55 patients. At the second ECT, patients receiving ECT at 150% above initial seizure threshold had significantly greater increases in plasma vasopressin than patients receiving low-dose ECT (ps < .01-.04), with no effects of electrode placement. At the second and ninth ECT treatments, the vasopressin or oxytocin surges were not associated with clinical improvement, seizure duration, time to orientation, or memory test performance. There were inverse trend-level associations between the acute surge in oxytocin levels at the ninth ECT and clinical response, contradicting a report in the literature. CONCLUSIONS: Overall, these findings do not support the hypothesis that diencephalic seizure propagation is central to the mechanism of action of ECT.