Molecular excitons in a copper azadipyrrin complex.

Exciton coupling is investigated in a copper azadipyrrin complex, Cu(L-aza)2. Exciton coupling in Cu(L-aza)2 assuming a single π-π* state on the L-aza ligand fails to account for the electronic structure of Cu(L-aza)2, which displays two almost equal intensity transitions at 15 600 cm(-1) and 17 690 cm(-1). TD-UB3LYP/6-31G(d) calculations suggest multiple π-π* transitions for the L-aza ligands and simple vector addition of the transition dipoles predicts two nearly orthogonal co-planar excitonic transitions that correctly reproduce the absorption band profile. Empirical modelling of absolute resonance Raman intensities using wavepacket dynamics confirms Cu(L-aza)2 has two equal intensity orthogonal exciton transitions. The phenyl substituents at the α- and γ-positions of the pyrrole rings play a central role in determining the orientation of the transition dipoles. Consequently the π-π* transitions for the L-aza ligands are oriented towards the substituent groups and are not in the plane of the pyrrole rings. Mode displacements in the Franck-Condon (FC) region obtained from the wavepacket model suggest that pyrrole ring and phenyl modes control the exciton FC dynamics. Our results suggest that Cu(L-aza)2 is an ideal model for theoretical, computational and experimental investigations of molecular excitons in molecular systems.

Vasotocin-like activity in the plasms of the euryhaline frog (Rana cancrivora) after transfer from fresh water to saline.

When Rana cancrivora were collected from fresh water and dehydrated (weight loss 4-10%) by exposure to saline, the plasma titre of hydro-osmotic activity, measured by amphibian bladder assay, was increased three-to fourfold. This activity, which was abolished by thioglycollate and by incubation with tyrosinase or trypsin, was ascribed to vasotocin. The plasma vasotocin activities (hydrated and dehydrated frogs respectively) were estimated to be 0-03-0-5 and 0-15-0-25 mug/1; if referred to oxytocin as a standard the equivalent values were 10 and 30-60 mu./ml. Assuming that the increase represented released pituitary hormone, the amount of vasotocin released by osmotic dehydration was calculated to be of the order of 1 ng. Pituitary glands of hydrated and dehydrated frogs were estimated to have 0.15 and 0-18 mug vasotocin/gland respectively. The possible physiological function of released vasotocin in promoting reabsorption of urea from the urinary bladder is discussed in relation to the euryhaline ability of R. cancrivora.

The effect of oxytocin on blood vessels of cremaster muscle of the rat.

Oxytocin administered intravenously to the anaesthetized rat produced dilatation of the blood vessels of the cremaster muscle at concentrations ranging from 2.5 times 10-minus 11 to 2.5 times 10-minus 9 M. When applied topically to the exposed vessels it produced constriction at concentrations ranging from 2.5 times 10-minus 12 to 5.0 times 10-minus 8 M. Oxytocin was thus similar to adrenaline in eliciting opposite effects when applied to the serosa or to the intima of skeletal muscle blood vessels.

Permeability of urinary bladder of Rana cancrivora to urea in the presence of oxytocin.

1. When Rana cancrivora collected from fresh water had been exposed for 3 days to saline solutions having osmolalities from 280 to 690 m-osmole/kg, urea concentrations in plasma and urine appeared to come into equilibrium, and were from 70 to 200 m-mole/l.2. Plasma urea level of fresh water R. cancrivora (48 m-mole/l.) was doubled (82 m-mole/l.) after 8 hr of exposure to 270 m-osmolal saline. It continued the same after 24 hr of exposure.3. When isolated urinary bladders of R. cancrivora were exposed to Ringer on the serosal aspect and one-fifth Ringer on the mucosal aspect, then in response to this osmotic difference of 190 m-osmole/kg, the rate of fluid movement (mucosa to serosa), which was 10.3(+/-2) mul./cm(2).hr, was not significantly altered when up to 60% of the NaCl of the Ringer solution was substituted by urea.4. Under the same circumstances, when oxytocin (50 m-u./ml.) was present in the serosal solution, the rate of fluid movement (mucosa to serosa) was 133.2(+/-7.9) mul./cm(2).hr in the absence of urea; it was progressively decreased by the presence of urea until, when 80% of the NaCl had been substituted by urea, the rate of fluid movement was reduced to 14.5(+/-4.0) mul./cm(2).hr.5. The diminished rate of fluid movement under the above circumstances could not be correlated with serosal urea concentration, with serosal availability of Na(+), nor with Na(+) concentration difference across the bladder wall. It appeared to be directly related to the ;non-urea osmotic difference' across the bladder wall provided by solutes other than urea.6. When isolated bladders were exposed to an osmotic difference of 190 m-osmole/kg, but having 25 mM urea present in the mucosal solution, then fluid moved from mucosa to serosa at a rate of 10.4(+/-1.3) mul./cm(2).hr in the absence of oxytocin and 124(+/-9) mul./cm(2).hr when oxytocin (50 m-u./ml.) was present. In the former case no urea passed across the bladder wall, but in the latter case urea passed from mucosa to serosa at a rate of 3.16(+/-0.3) mumole/cm(2).hr. The fluid moving from mucosa to serosa thus contained urea 25.5 m-mole/l.7. Vasotocin (10(-9)M), which is equipotent with oxytocin (50 m-u./ml.) in affecting permeability of the isolated urinary bladder to water, was also equipotent in producing a reduced rate of water fluid movement in the presence of 40% urea (vasotocin, 63 mul./cm(2).hr; oxytocin, 59 mul./cm(2).hr).8. When groups of frogs were cystectomized, and other groups of frogs were sham-operated, then after 48 hr of exposure to fresh water or to 300 m-osmolal saline the sham-operated frogs had plasma urea level raised from 20 m-mole/l. (fresh water) to 42 m-mole/l. (saline), while the cystectomized frogs had 20 m-mole/l. (fresh water) and 26 m-mole/l. (saline).9. The hypothesis is presented that hormone-induced permeability of the urinary bladder to urea contributes to the immediate adjustment of plasma urea level by which R. cancrivora survives when exposed to high environmental salinity.

Effect of neurohypophysial hormones on fluid movement across isolated bladder of Rana cancrivora, Rana temporaria and Bufo melanostictus.

1. The rate of fluid movement across the wall of isolated amphibian urinary bladder was estimated using hemi-bladders of the tropical euryhaline Rana cancrivora, the temperate R. temporaria and the tropical Bufo melanostictus.2. The rate of water outflow across the bladder walls was: Rana cancrivora, 4.8 mul./cm(2).hr; R. temporaria, 7.2 mul./cm(2).hr; Bufo melanostictus, 3.7 mul./cm(2).hr.3. When neurohypophysial peptides were added to the serosal solution the rates of water outflow were increased; dose-response curves were prepared relating the augmented rate of fluid movement to the concentration in the bathing fluid of vasopressin, oxytocin or vasotocin.4. The potencies of the peptides in respect of bladders of each species were:Vasopressin, B. melanostictus:Rana temporaria:R. cancrivora, 500:20:1.Oxytocin, R. temporaria:B. melanostictus:Rana cancrivora, 100:5:1.5. The ratio of sensitivity of the bladder of each species to the presence of the different peptides was:R. cancrivora, vasotocin:oxytocin:vasopressin, 1000:5:1.R. temporaria, oxytocin:vasopressin, 15:1. Bufo melanostictus, oxytocin:vasopressin, 1:30.6. The effect of extract of the pituitary gland of Rana cancrivora was examined on R. cancrivora bladder. It had a dose-response curve similar to the dose-response curve of vasotocin.7. The vasotocin content of the pituitary gland of R. cancrivora was estimated(a) from the dose-response curves using R. cancrivora bladder,(b) by a three-point assay using R. cancrivora bladder and(c) by its effect on water uptake across the skin of Bufo melanostictus. One pituitary gland of Rana cancrivora was found to contain about 0.1 mug vasotocin.8. The urinary bladder of R. cancrivora was less responsive to neurohypophysial peptides than the bladder of R. temporaria or Bufo melanostictus.9. The hormonal content of the pituitary gland of Rana cancrivora was smaller than that found in other amphibians.

Water uptake by the crab-eating frog Rana cancrivora, as affected by osmotic gradients and by neurohypophysial hormones.

1. The rate of water uptake across the skin was investigated in live Rana cancrivora, an euryhaline frog which has been reported to tolerate sea water. When they were exposed to distilled water at 29 degrees C, the rate of water uptake was 8.4 +/- 0.4 mul./cm(2).hr; when bathed in solutions ranging from 30 to 570 m-osmole/l., irrespective of whether the solute was sucrose, urea or NaCl, the rate of fluid uptake during the first day was inversely related to the osmolarity of the solution. No appreciable fluid movement was observed when the bathing solution had an osmolar concentration of 270 m-osmole/l.2. The rate of fluid uptake was not affected by injections of vasopressin, oxytocin or of extracts of amphibian or rat pituitary glands, irrespective of whether R. cancrivora were bathed in distilled water or in solutions of NaCl or sucrose.3. In Bufo melanostictus, in contrast with R. cancrivora, injections of neurohypophysial extracts produced a marked increase of the rate of fluid uptake.4. In the laboratory, R. cancrivora could be acclimatized stepwise to tolerate NaCl solutions up to 700 m-osmole/l. for 7 days.5. After 24 hr exposure either to distilled water or to NaCl solutions from 100 to 670 m-osmole/l., the osmolar concentration of the plasma of R. cancrivora was always higher than that of the bathing fluid. In R. pipiens or R. temporaria plasma osmolar concentration was higher than that of the bathing fluid only when the latter did not exceed 300 m-osmole/l.6. Under all conditions investigated, the osmolar concentration of the urine of R. cancrivora was always lower than that of the plasma.7. The amounts of pressor and oxytocic activities of pituitary glands of R. cancrivora kept in distilled water or in NaCl solutions up to 300 m-osmole/l. were 8.9 +/- 0.8 and 1.8 +/- 0.3 m-u./gland, irrespective of sex or body weight within the range 30-50 g. After 3 days exposure to hypertonic NaCl solutions, the amounts of pressor and oxytocic activities were 14.7 +/- 1.2 and 3.1 +/- 0.3 m-u./gland. In both instances the pressor/oxytocic ratio was 4.9. Pituitary glands of R. temporaria similarly showed increased pressor and oxytocic activities after exposure to NaCl solutions of 300-360 m-osmole/l.

Natriferic and hydrosmotic effects of neurohypophysial peptides and their analogues in augmenting fluid uptake by Bufo melanostictus.

1. The effect of intramuscular injection of 8-arginine vasotocin, 8-arginine vasopressin, 8-lysine vasopressin, oxytocin, 8-ornithine oxytocin and 8-ornithine vasopressin on fluid uptake across the skin was studied in the live toad, Bufo melanostictus, bathed either in distilled water or in NaCl solution (0.1 g/100 ml.).2. When the bathing solution was distilled water, 8-arginine vasotocin was the most potent, 0.14 nmole/kg augmenting the rate of fluid uptake by 50%. Compared with it the others had relative potencies of: 8-arginine vasopressin 0.8, 8-lysine vasopressin 0.8 x 10(-3), oxytocin 0.8 x 10(-3), 8-ornithine oxytocin 0.8 x 10(-2), 8-ornithine vasopressin < 1.4 x 10(-4).3. When the bathing solution contained 0.1% NaCl, 8-arginine vasotocin was again the most potent, 0.06 nmole/kg augmenting the rate of fluid uptake by 50%. Compared with it the others had relative potencies of: 8-arginine vasopressin 0.3, 8-lysine vasopressin 0.3 x 10(-3), oxytocin 0.3 x 10(-2), 8-ornithine oxytocin 0.8 x 10(-2), 8-ornithine vasopressin < 0.6 x 10(-4).4. Dose-response curves for each peptide showed that in the case of 8-arginine vasopressin, 8-lysine vasopressin and 8-ornithine vasopressin the augmentation of rate of fluid uptake did not differ in the absence or in the presence of NaCl in the bathing solution; whereas in the case of 8-arginine vasotocin, oxytocin, and 8-ornithine oxytocin the augmentation was greater in the presence of sodium chloride.5. Support has been found for the postulate of a binary action of some neurohypophysial peptides on amphibian skin, arginine in position 8 being correlated with hydrosmotic effect, and isoleucine in position 3 with natriferic effect.

Water uptake by Bufo melanostictus, as affected by osmotic gradients, vasopressin and temperature.

1. The rate of water uptake across the skin was studied in the live toad, Bufo melanostictus. When toads were kept in distilled water at 29 degrees C the uptake of water amounted to 16.9 +/- 1.3 mul./cm(2)/hr; when bathed in sucrose or urea solutions, the water uptake diminished with increasing osmotic pressure. There was no water uptake observed when toads were kept in 200 m-osmolar sucrose or urea.2. Intramuscular injections of vasopressin increased the rate of water uptake from distilled water. There was a good relation between doses and responses over various time intervals. A dose of 4 m-u. vasopressin/g body wt. doubled the rate of water uptake over a period of 1 hr. The same dose of vasopressin doubled the rate of water uptake when the toads were kept in solutions of sucrose or urea of different osmolarity.3. The rate of water uptake when the toads were bathed in sodium chloride solutions was consistently 8 mul./cm(2)/hr greater than when bathed in sucrose or urea solutions of equal osmolarity. There was no water uptake when the sodium chloride solution was 285 m-osmolar.4. Vasopressin (4 m-u./g) injected intramuscularly doubled the rate of water uptake from sodium chloride solutions of different osmolarity.5. With solutions of potassium chloride, sodium nitrate, and potassium nitrate, in concentrations up to 150 m-osmoles/l., the rate of water uptake was found to be the same as with solutions of sodium chloride of the same osmolarity. Similarly, it was doubled by injection of vasopressin (4m-u./g).6. The effect of temperature on the rate of water uptake before and after injection of vasopressin was investigated in toads kept in distilled water, sucrose, or sodium chloride solutions. For temperatures between 20 and 37 degrees C, vasopressin (4 m-u./g) reduced the activation energy involved in the process of water uptake by 4000 cal.7. The results agree with the view that water uptake follows a diffusion process which is facilitated by vasopressin, possibly as a result of increasing the size or number of available pores.