Effect of nitroprusside on smooth muscle and adrenergic nerve terminals in isolated blood vessels.

Experiments were designed to assess the mode of action of nitroprusside on isolated blood vessels and its relative potency on venous and arterial smooth muscle. Strips from dog blood vessels were mounted in an organ bath for isometric tension recording. Sodium nitroprusside (10(-5) M) depressed the contraction of saphenous vein strips caused by electric stimulation, tyramine, K+, Ba++, norepinephrine and acetylcholine. The depression of the norepinephrine-induced contractions also occurred in a Ca++- free medium and when Ca++ influx was inhibited by verapamil. Nitroprusside reduced the frequency of the spontaneous contractions of strips of portal-mesenteric veins. It depressed the contraction caused by norepinephrine in tibial artery strips more than in saphenous vein strips. Saphenous vein strips were incubated with (3H)norepinephrine and mounted for superfusion and isometric tension recording. Sodium nitroprusside (10(-5) M) had no effect on the basal efflux of 3H compounds. During electric stimulation, it did not change the output of (3H)norepinephrine but increased the outflow of deaminated and O-methylated metabolites. Thus sodium nitroprusside 1) has a direct effect on the smooth muscle cells which is independent of Ca++ influx, 2) depresses contractions of different types of vascular smooth muscle and 3) does not inhibit the release of norepinephrine from the nerve endings.


Metabolic activation of furosemide to a chemically reactive, hepatotoxic metabolite.

The possibility that furosemide-induced hepatic necrosis results from the formation of a chemically reactive hepatotoxic metabolite has been examined. Hepatotoxic doses of 3H-furosemide or 14C-furosemide were administered to normal mice and to mice pretreated with piperonyl butoxide, cobalt chloride, alpha-naphthylisothiocyanate or phenobarbital. Mice were killed at various time intervals and tissues were examined for necrosis, for free furosemide concentrations and for covalently bound metabolites of furosemide. Little furosemide was covalently bound to muscle, whereas the amount of covalently bound material in liver usually paralleled the severity of live necrosis after alteration by the pretreatments. The severity of hepatic necrosis failed to correlate with furosemide concentrations in liver or plasma. Furosemide was shown to be metabolically activated to an arylating intermediate by a cytochrome P-450 mixed function oxidase in hepatic microsomes. Additional experiments demonstrated that the furan ring of furosemide was the portion activated.


Comparison of the effect of carbon monoxide and of hypoxic hypoxia. I. In vivo metabolism, distribution and action of hexobarbital.

Previous experiments had shown that carbon monoxide inhalation prolonged the in vivo response zoxazolamine. However, CO was clearly less potent than an equivalent level of hypoxia induced by inhalation of lowered O2 content (hypoxic hypoxia). The present study revealed that both CO and hypoxic hypoxia also prolonged the pharmacologic response to a drug of vastly different structure and site of action, i.e., hexobarbital. In this case, CO was approximately equipotent to hypoxic hypoxia in prolonging sleeping time. In spite of a number of significant effects of the two types of hypoxia on factors such as rate of peritoneal absorption, distribution into the brain and brain sensitivity to the drug, their relative potencies in prolonging sleeping time appeared to reflect adequately their relative effects on in vivo rate of metabolism. This conclusion was based on their equipotencies in decreasing the rate of disappearance of hexobarbital from the blood. The observation that hypoxia induced by CO had no greater effect on hexobarbital metabolism than hypoxia induced by oxygen deprivation suggests that the binding of CO to cytochrome P-450 is not important in drug metabolism in vivo. CO would appear to act solely by inducing tissue hypoxia.


Comparison of the effect of carbon monoxide and of hypoxic hypoxia. II. Hexobarbital metabolism in the isolated, perfused rat liver.

Rat livers were perfused with a red blood cell-containing perfusion medium. The rate of metabolism of hexobarbital was determined in this system as the recirculating perfusion fluid was made hypoxic by exposure to either CO or lowered oxygen tension. It was found that 7.6% of the total oxygen consumed by the liver was utilized to metabolize hexobarbital regardless of the severity of the hypoxia, which suggests that the liver does not shunt O2 to more preferential pathways as O2 availability becomes limited. When oxygen delivery was lowered below 0.3 ml of O2 per min per g of liver by either type of hypoxia, the rate of hexobarbital metabolism decreased. However, based on the rate of oxygen delivery, CO was more potent than hypoxic hypoxia in inhibiting drug metabolism. On the other hand, the two types of hypoxia were equipotent when compared on the basis of alterations in oxygen consumption. These results are consistent with the known effect of CO in decreasing the availability of hemoglobin-bound O2 and further argue against the direct inhibition of drug metabolism by the binding of CO to cytochrome P-450 in the isolated liver. They also suggest that the lack of a greater potency of CO relative to hypoxic hypoxia in vivo might be related to alterations in hepatic blood flow.


Studies of the metabolism of 5,5-diphenylhydantoin relating principally to the stereoselectivity of the hydroxylation reactions in man and the dog.

The hydroxylated metabolites of 5,5-diphenylhydantoin (DPH) in dog and human urine, after release by beta-glucuronidase, have been isolated and purified by procedures entailing only solvent partitioning without crystallization or any other procedure that could change the proportions of optical isomers. Dogs produced both the meta- and para-5-hydroxyphenyl-5-phenylhydantoins (m-HPPH and p-HPPH) from DPH, the former in approximately 6 times the amount of the latter. The m-HPPH from dog urine was dextrorotatory and had the properties of a pure optical isomer. The p-HPPH from dog urine was a mixture of optical isomers with approximately a 2:1 preponderance of the levorotatory isomer. When dog urine was heated with acid rather than treated with beta-glucuronidase, the isolated p-HPPH contained a small preponderance of the dextrorotatory isomer, probably owing to production of d-p-HPPH from the dihydrodiol metabolite. No o-HPPH was found in dog urine. When racemic p-HPPH and m-HPPH were administered to dogs, there was little if any preponderance of either optical isomer in the materials isolated from urine. Enzymatically released p-HPPH isolated from urine of human patients receiving DPH was a mixture of optical isomers with approximarely a 10:1 preponderance of the levorotatory isomer. Crystallization of this material readily yields pure l-p-HPPH. Human urine contained only very small amounts of m-HPPH, which was detectable by gas chromatography but insufficient in amount to permit isolation. Hypotheses are discussed that could account for the differences in the metabolism of DPH in man and dog. Small amounts of 2,2-diphenylhydantoic acid were found in urine of dogs but not of human patients receiving DPH.


Role of metabolism in the biliary excretion of methadone metabolites.

The biliary excretion of the major methadone metabolites was studied after the administration of purified 14C-labeled metabolites of methadone. The three methadone metabolites administered to rats were the pyrrolidine and pyrroline metabolites and the glucuronide conjugate of the hydroxylated pyrroline metabolite. We found that the excretion of 14C into bile was greater after administration of the metabolites which underwent fewer metabolic steps prior to biliary excretion. Phenobarbital pretreatment of rats increased the biliary excretion of 14C after the administration of the 14C-labeled pyrrolidine and pyrroline metabolites, both of which undergo further metabolism. Phenobarbital pretreatment did not change the biliary excretion of 14C-labeled glucuronide of the pyrroline metabolite which does not undergo further metabolism in the liver. Analysis of bile samples showed that the increased biliary excretion of 14C in phenobarbital-pretreated rats after administration of the labeled pyrrolidine and pyrroline metabolites was due primarily to increases in the biliary excretion of methadone metabolites which resulted from further in vivo metabolism of the injected metabolites. These data confirm our earlier suggestion (Roerig et al., Biochem. Pharmacol. 24: 355-362, 1975) that phenobarbital pretreatment increases biliary excretion of methadone metabolites by increasing hepatic metabolism of methadone. Furthermore, it can be concluded from these studies that metabolism is a rate-limiting process in the excretion of methadone metabolites in bile.


Ischemic changes in the canine heart as affected by the dimethyl quaternary analog of propranolol, UM-272 (SC-27761).

The effects of the dimethyl quarternary analog of propranolol, UM-272, on myocardial infarct volume were studied in the canine heart. Myocardial infarction was produced by occlusion of the left circumflex coronary artery for 60 minutes followed by reperfusion and quantitation of infarct volume 24 hours later. Groups of dogs were either untreated or pretreated with UM-272 with an initial loading dose of 5.0 mg/kg (group A) or 2.5 mg/kg (group B) 30 minutes before occlusion of the left circumflex coronary artery. Both group A and group B animals received additional doses of 2.5 mg/kg of UM-272 every 90 minutes for a period of 6 hours so that the total respective doses were 15 and 12.5 mg/kg. Control animals received comparable volumes of 0.9% sodium chloride solution. All animals were followed throughout the 6-hour procedure with continuous electrocardiographic recordings which were used to assess the effects of acute myocardial ischemia upon disturbances in cardiac rhythm and the effects of drug treatment. Dogs which survived the procedure were given tetracycline i.v. the next day and sacrificed 1 hour later by an overdose of pentobarbital sodium. The hearts were removed and the left ventricle was sliced and examined first under ultraviolet light to localize the ischemic zone by noting the tetracycline fluorescence. The ventricular slices were next incubated in nitro blue tetrazolium which stains normal myocardial tissue, thus allowing one to quantitate the volume of infarcted myocardium by excising and weighing the nonstained and stained muscle separately. The untreated control group had an infarct volume of 23.8 +/- 3.2 g/100 g of left ventricle. The treated animals in groups A and B had respective infarct volumes of 2.3 +/- 0.8 g/100 g (P less than .001) and 7.0 +/- 3.3 g/100 g (P less than .025) of left ventricle. During the acute phase of ischemia and reperfusion, arrhythmias and alterations in the ST-segment, R-wave amplituted and development of pathologic Q-waves were more prominent in the untreated animals and almost totally absent in the treated animals. UM-272 produced a dose-dependent decrease in heart rate as well as a decrease in developed isometric tension. Pretreatment with UM-272 did not prevent the derangement of function in the ischemic zone nor did it permit a return of function upon reperfusion, even though it reduced the degree of cellular damage resulting from 60 minutes of regional ischemia. A possible mechanism for the protective effect of UM-272 may be through its ability to reduce myocardial contractility and heart rate, both of which would reduce myocardial oxygen consumption and thus produce a more favorable balance between myocardial oxygen supply and myocardial oxygen demand.


Phentolamine and isoproterenol: comparison of effects on vascular resistance and oxygen uptake in skeletal muscle during hypotension.

Effects of drugs on precapillary vessels and oxygen uptake may differ from effects on resistance vessels and total flow in skeletal muscle. This study was performed to compare effects of phentolamine and isoproterenol, two drugs which are used to treat shock, on oxygen uptake in skeletal muscle. Oxygen uptake of gracilis muscle (GVO2) was measured in muscles perfused at constant flow or constant pressure in normotensive dogs. We compared effects of the drugs on oxygen uptake at doses chosen so that both drugs produced comparable effects on vascular resistance. With flow constant, phentolamine increased but isoproterenol decreased or did not alter GVO2. With constant pressure, phentolamine produced significantly greater increases in GVO2. For example, increases in GVO2 occurred with all three doses of phentolamine, but only with the high dose of isoproterenol. Neither drug altered oxygen-hemoglobin affinity of red blood cells or oxygen consumption of skeletal muscle in vitro. The results suggest that phentolamine produces more favorable effects than isoproterenol on oxygen uptake in skeletal muscle, presumably because of greater dilator action on precapillary sphincters.


Uptake, metabolism and efflux of methadone in "single pass" isolated perfused rabbit lungs.

The uptake, metabolism and persistence of racemic, d- and l-methadone were studied in the isolated perfused rabbit lung. The removal of the drug from the perfusate was resolved into two uptake processes (I1 and I2Y. The unidirectional flux of methadone from the perfusate into the lung was linear with respect to perfusate concentration, suggesting an uptake mechanism involving diffusion and/or binding. The uptake of d- and l-methadone was identical. In the absence of methadone in the perfusate, methadone that previously had accumulated in the lung effluxed at three rates (T1/2 = 0.37, 1.65 and 8.9 minutes) suggesting that accumulated methadone was stored in at least three pools (E1, E2, and E3) In addition, a pool with a half-life in excess of 5 hours (noneffuxable pool) was detected; this noneffluxable pool was shown not to be the result of irreversible covalent binding. Methadone was biotransformed to a small extent by the isolated perfused lung to mono- and di-N-demethylated metabolites. Although unchanged methadone and its N-demethylated metabolites were consistently found in the lung at the end of efflux, the mono-N-demethylated metabolite was not always detectable in the perfusate. Nevertheless, the decrease in the rate of efflux of the metabolites with respect to time was identical to that of unchanged methadone.


Effects of methadone hydrochloride on the growth of organotypic cerebellar cultures prepared from methadone-tolerant and control rats.

Male and female Sprague-Dawley rats were given dl-methadone (5 mg/kg) for at least 3 months and then mated. The drug was continued throughout pregnancy and after delivery. The newly born pups were divided into two groups. One group was tested for in vivo methadone tolerance, while the animals in the othergroup were used to prepare organotypic cerebellar cultures. Various amounts of dl-methadone were added to the media of half of these cerebellum cultures. The effect of the drug in the medium was assessed by measuring explant outgrowth. Similar experiments were carried out with control animals. Statistical analysis of the data obtained in the in vivo portion of the experiment indicates that the pups of methadone-treated mothers tolerate methadone better than those of untreated mothers. The culture experiments revealed that the addition of methadone to the medium reduced explant outgrowth size and this was a dose-related effect. Also, there was significantly less outgrowth from explants prepared using pups of methadone-treated mothers as compared to the controls. There was no significant difference in the effect of methadone on the growth of cultures prepared from the methadone-tolerant and control animals.


Naloxone blockade of morphine analgesia: a dose-effect study of duration and magnitude.

The purpose of the experiment was to determine the minimal dose of naloxone needed to block the analgesic effect of morphine as measured by the foot-shock titration procedure in the rat. Various groups of rats received, subcutaneously, various doses of naloxone hydrochloride, from 0.03 to 2.0 mg/kg, 15 minutes before receiving 10 mg/kg of morphine sulfate. The results indicate that 1.0 mg/kg of naloxone is the minimum dose necessary to provide full blockade of the measured morphine effect for up to 5 hours after the morphine administration. Partial blockade was evident at doses much smaller than 1.0 mg/kg of naloxone. A dose of 0.125 mg/kg of naloxone completely blocked the onset of a measurable effect from 10 mg/kg of morphine for 60 minutes in six out of the seven animals tested at that dose. The results provide a clear dose-effect relationship for both magnitude and duration of naloxone blockade of the analgesic effect of morphine.


Effect of l-propoxyphene on plasma levels and analgesic activity of d-propoxyphene in the rat.

Oral administration of l-propoxyphene with d-propoxyphene enhances the analgesic activity of d-propoxyphene as expressed in the rat tail heat test. The combination of d- and l-propoxyphene at a dose of 10 mg/kg each was found to have activity in the analgesic assay equivalent to that observed with d-propoxyphene at a dose of 20 mg/kg. In the same test, l-propoxyphene at a dose of 40 mg/kg had no activity. Co-administration of equal amounts of l-propoxyphene with d-propoxyphene (10 mg/kg p.o.) results in an increase in circulating plasma levels of d-propoxyphene from 9 +/- 2 to 114 +/- 39 ng/ml 15 minutes after administration. The increase in plasma levels is accompanied by a proportional increase in the brain and lung levels with no significant change in the liver levels. When d-propoxyphene (4 mug/ml) was infused in the isolated perfused rat liver, over 98% of the drug was extracted in a single pass through the liver. When l-propoxyphene was added to the perfusate (4 mug/ml), the extraction of d-propoxyphene was decreased to less than 90%. These results indicate that l-propoxyphene increases the systemic availability of d-propoxyphene by altering the amount of d-propoxyphene extracted by the liver.


Disposition of dobutamine in the dog.

The disposition of dobutamine, a potent inotropic catecholamine, has been examined in the dog. A sensitive assay has been developed for dobutamine allowing the measurement of the drug at plasma concentrations as low as 1 ng/ml. The short plasma half-life of dobutamine from the plasma to the tissue. When 14C-dobutamine was administered, the plasma half-life of radioactivity was 1.9 hours. The circulating radioactivity consists mainly of the glucuronide conjugate of 3-O-methyldobutamine. During continuous intravenous administration of dobutamine, plasma levels of metabolites reached a maximum between 3 and 4 hours. During a 48-hour time period after administration of 14C-dobutamine, 67% of the radiolable was excreted in the urine and 20% in the feces. In dogs having cannulated bile ducts, 30 to 35% of the administered drug was excreted in the bile. The major urinary metabolites were the glucuronide conjugates of dobutamine and 3-O-methyldobutamine.


Ionic mechanism of the salicylate block of nerve conduction.

Benzoic acid, salicylic acid and 5-bromosalicylic acid have been studied for their mechanism of action on crayfish and squid giant axons. External application of 0.1 to 0.5 mM 5-bromosalicylic acid prolonged the falling phase of the action potential, and at concentrations higher than 1.5 mM, nervous conduction was blocked. Boltage clamp analyses showed that 5-bromosalicylic acid reduced both peak transient sodium conductance and steady-state potassium conductance. The curves relating the peak and steady-state conductances to membrane potential were shifted in the direction of depolarization. The time constant for the activation of sodium current (taum) was not affected by 5-bromosalicylic acid, whereas the time constant for the sodium inactivation (tauh) was greatly increased. The steady-state sodium inactivation curve (h infinity) was shifted in the hyperpolarizing direction. The rate constants alpha h and beta h were decreased. The rate of activation of potassium current was reduced. The block of peak transient conductance by externally applied 5-bromosalicylic acid was largely reversed by internal washing suggesting that the drug acts on a membrane site more easily accessible from inside than from outside. Salicylic and benzoic acids were much less potent than 5-bromosalicylic acid, reducing peak conductance by 12.5 and 3%, and steady-state conductance by 8.7 and 7%, respectively, at a concentration of 10 mM.


The magnitude and duration of respiratory depression produced by fentanyl and fentanyl plus droperidol in man.

In 10 healthy male volunteers breathing 100% oxygen, we determined the effect of four intravenous dose levels of fentanyl (0.0015, 0.003, 0.006 and 0.009 mg/kg) and two of fentanyl plus droperidol (i.e., Innovar, 0.003 and 0.006 mg/kg of fentanyl with 2.5 mg of droperidol for each 0.05 mg of fentanyl) on PECO2 and the slope of the ventilatory response to imposed increases in PECO2. All doses of fentanyl and fentanyl plus droperidol depressed the slope and shifted the curve to the right. Depression was dose related and was maximum 5 minutes after administration. The slope returned to control by 2 hours postinjection even at the highest narcotic dose. However, the rightward shift of the CO2 response curve require 4 hours to return to control. Droperidol added to fentanyl did not increase or prolong the respiratory depression seen with fentanyl alone at equivalent dose levels. Nausea and emesis occurred more frequently with fentanyl alone and orthostatic hypotension occurred more frequently with droperidol plus fentanyl. Dysphoria was a prominent consequence of fentanyl plus droperidol administration.


Co-transport of glycine and sodium across the mucosal border of the midgut epithelium in the marine shrimp, Penaeus marginatus.

1. Influxes of glycine and Na across the mucosal border of the epithelial cells lining the midgut lumen of a marine shrimp have been examined. 2. Both mucosal JG and JNa displayed Michaelis-Menten kinetics and the influx of each solute was markedly dependent upon the presence of the other solute in the bathing solution. A portion of JG occurred in the absence of Na, but JNa appeared to be completely dependent upon glycine. 3. Glycine influx Kt and Jmax were dependent upon [Na] while Na influx Kt and Jmax depended upon [glycine]. 4. K acted as a competitive inhibitor of Na binding to the co-transport carrier by altering the Kt of glycine influx, but was without significant effect on the amino acid Jmax. 5. The coupling coefficient of mucosal co-transport (JNa/JG) varied with the [Na] being 1-1 at low [Na] (471 mM). 6. A model is proposed in which Na and glycine share ternary (NaXG) and quaternary (Na2XG) carrier complexes during their coupled influxes, but in which glycine may also traverse the mucosal membrane alone by way of an unshared binary (XG) carrier complex.


Heat production and chemical change during isometric contraction of rat soleus muscle.

1. Methods are described whereby the soleus muscle of the rat may be used for the investigation of initial processes in the absence of oxidative recovery. 2. The anaerobic conditions employed had no effect on the concentration of phosphocreatine in resting muscle or the mechanical response during contraction. 3. Muscles were stimulated tetanically for 10 s at 17-18 degrees C. Measurements were made of the heat production and metabolic changes that occurred during a 13 s period following the first stimulus. 4. There was no detectable change in the concentration of ATP. Neither was there detectable activity of adenylate kinase or adenylate deaminase. The changes in the concentration of glycolytic intermediaries were undetectable or very small. 5. The change in the concentration of phosphocreatine was large and amounted to -127 +/- 11-4 mumol/mmol Ct (mean and S.E. of the mean, negative sign indicates break-down, Ct = free creatine + phosphocreatine) which is equivalent to about -2-13 mumol/g wet weight of muscle. The heat production was 6549 +/- 408 mJ/mmol Ct (mean and S.E. of mean) which is equivalent to about 110 mJ/g. 6. About 30% of the observed energy output is unaccounted for by measured metabolic changes. 7. The ratio of heat production (corrected for small amounts of glycolytic activity) to phosphocreatine hydrolysis was -49-7 +/- 5-6 kJ/mol (mean and S.E. of mean), in agreement with previous results using comparable contractions of frog muscle, but different from the enthalpy change associated with phosphocreatine hydrolysis under in vivo conditions (-34 kJ/mol). 8. The results support the notion that the discrepancy between energy output and metabolism is an indication of an unidentified process of substantial energetic significance that is common to a number of species.


Properties of motor units of the frog sartorius muscle.

1. The mechanical properties of single motor units in the sartorius muscle of the frog Litoria aurea were examined during single shock and repetitive stimulation of motor axons. 2. The tetanic tension developed by motor units lay in the range 1-40% of whole muscle tension with two peaks in the distribution, in the range 5-10% and 25-30%. The large units had briefer times-to-peak for the twitch than the small units and were more readily fatigued during prolonged repetitive stimulation. 3. Histological examination of the muscle gave a count of 620 muscle fibres with a diameter range of 28-128 mum. Cholinesterase stained preparations showed that the majority of muscle fibres had several nerve terminals (mean 3, range 1-5). 4. Muscle fibres received their multiple innervation from different axons (polyneuronal) or branches of the same axon (multiterminal). The presence of polyneuronal innervation of muscle fibres was confirmed by a comparison of the tensions when each of a pair of motor units was stimulated alone and when they were stimulated together. The tension excess, or overlap, was up to 60% when expressed in terms of the tension developed by either unit alone. Motor units developing similar amounts of tension tended to show more overlap in their innervation than units with very different tensions. 5. An estimate of the amount of multiterminal innervation gave variable results but could account for up to 60% of a motor unit's tension. No correlation could be detected between the values for multiterminal innervation and any other measured parameter. However, it is argued that because of the limitations of the measurements the existence of a relationship between the extent of multiterminal or polyneuronal innervation and the mechanical properties of the motor unit cannot be excluded.


The reduced responsiveness of neurones in nucleus reticularis gigantocellularis following their excitation by peripheral nerve stimulation.

1. Post-stimulus histograms of neuronal activity, constructed from extracellular recordings in decerebrate, decerebellate cats, have been used to investigate the responsiveness of neurons in nucleus reticularis gigantocellularis following their excitation by a peripheral nerve stimulus. 2. The response to a testing stimulus applied to a peripheral nerve was depressed following the response to a conditioning stimulus applied to the same or a different peripheral nerve. This reduction in responsiveness was maximal within 50 msec of the peak of the response to the conditioning stimulus. Response latencies to the testing stimulus were increased during the period of reduced responsiveness. 3. Responsiveness to a peripheral nerve stimulus was also reduced following a spontaneous or an antidromically evoked spike, but this effect was weaker and much shorter-lasting than that following a nerve-evoked spike. Thus, the reduced responsiveness cannot be solely due to phenomena which are an inevitable consequence of an action potential in the neurone. 4. In spontaneously firing neurones, the duration of the reduced responsiveness to a testing stimulus generally outlasted the depression of spontaneous activity which often followed an excitation evoked by a peripheral nerve conditioning stimulus. 5. The reduction in responsiveness to a testing stimulus applied to the same nerve as the conditioning stimulus was greater and longer-lasting than that to a testing stimulus applied to a different nerve. 6. When stimuli were applied to one nerve at a relatively high rate, the neurone became much less responsive to that input, but simultaneously became more responsive to low rate stimulation of other nerves. 7. It is concluded that the greater part of the reduced responsiveness is due to events occurring on the input pathway to a reticular neurone, or possibly in the region of the afferent endings on its dendrites. These processes may allow selective changes in responsiveness to different inputs, and enable the units to act as novelty detectors.


Growth of rats' kidneys after unilateral uretero-caval anastomosis.

1. The relative significance of reduced excretion of urinary constituents and reduced renal mass, as stimuli to growth of one kidney after the other has been removed, has been investigated. 2. To abolish the excretory function of one kidney without removing it, the right ureter was drained into the vena cava through a compound cannula for 6 weeks. Uretero-caval anastomoses were performed in twenty-four male rats at 10 weeks of age: six survived without evidence of ureteric obstruction (and a further five with minimal obstruction). 3. The rats with anastomoses grew less than six other rats from which the right kidney had been removed or six which had been submitted to a sham operation, and their plasma urea and creatinine concentrations were higher. 4. Relative to body weight, the dry weight of each kidney after uretero-caval anastomosis without obstruction was 18% greater than after sham operation; taking both kidneys together, the total increase was almost as much as in the left kidney alone after right nephrectomy (46%). 5. Histologically and in terms of DNA concentration, the growth of both kidneys after uretero-caval anastomosis was of the same kind as in the left kidney after right nephrectomy. 6. The return of urine from one kidney into the blood provided a powerful stimulus to renal growth in spite of the restraining effect of increased renal mass.


Temporal response patterns of the human sinus node to brief carotid baroreceptor stimuli.

1. A new technique, timed, quantitative neck suction was used to establish in normal man the temporal response patterns of the sinus node to brief baroreceptor stimuli. 2. Maximum sinus node inhibition occurred when stimuli were delivered about 0-75 sec before the anticipated appearance of the subsequent P wave; less pulse interval prolongation occurred when stimuli were delivered earlier, or later in the cardiac cycle. 3. The timing of maximum sinus node responsiveness appeared to be independent of base-line cardiac cycle length. 4. The duration of the baroreceptor-cardiac reflex are (the time from the onset of baroreceptor stimulation until the earliest occurrence of pulse interval prolongation) averaged 0-24 sec. 5. Pulse interval prolongation was proportional to baroreceptor stimulus strength and duration. 6. The results suggest that in man, sinus node responsiveness to baroreceptor stimuli depends critically upon the timing of stimuli within the cardiac cycle.


Integrated electromyogram and oxygen uptake during positive and negative work.

1. Integrated electromyogram (e.m.g.) from the vastus lateralis muscles, and steady-state rates of oxygen uptake, were measured simultaneously during the performance of set rates of positive (concentric) and negative (eccentric) work at 50 rev/min on a motorized bicycle ergometer. 2. Similar experiments were also carried out at other pedalling rates and using other leg muscles. 3. The relationships between each of the variables (integrated e.m.g., oxygen consumption) and mean torque on pedals were found to be linear (r greater than 0-98) with a remarkable degree of reproducibility in surface e.m.g. for each subject over several months. 4. The ratio of the e.m.g. slopes at 50 rev/min (positive/negative) was 1-96 +/- 0-12 while the same ratio for the oxygen uptake slopes was 6-34 +/- 0-82. The discrepancy between the ratios suggests that not only is less muscle fibre activity required to maintain the same exerted force during negative work exercise, but there is also a substantial reduction in the oxygen uptake when the fibres are stretched. This was observed for all speeds of pedalling.


Spike initiation by transmembrane current: a white-noise analysis.

1. Those features of a transmembrane current correlated with spike initiation were examined in Aplysia neurones using a Gaussian white-noise stimulus. This stimulus has the advantages that it presents numerous wave forms in random order without prejudgement as to their efficacies, and that it allows straightforward statistical calculations. 2. Stimulation with a repeating segment of Gaussian white-noise current revealed remarkable invariance in the firing times of the tested neurones and indicated a high degree of reliability of their response. 3. Frequencies (less than 5 Hz) involved in spike triggering propagated faithfully for up to several millimetres, justifying intrasomatic current injection to examine spike initiation at the trigger locus. 4. Examination of current wave forms preceding spikes indicated that a wide variety could be effective. Hence, a statistical analysis was performed, including computation of probability densities, averages, standard deviations and correlation coefficients of pairs of current values. Each statistic was displayed as a function of time before the spike. 5. The average current trajectory preceding a spike was multiphasic and depended on the presence and polarity of a d.c. bias. An early relatively small inward- or outward-going phase was followed by a large outward phase before the spike. The early phase tended to oppose the polarity of the d.c. bias. 6. The late outward phase of the average current trajectory reached a maximum 40--75 msec before triggering the action potential (AP) and returned to near zero values at the moment of triggering. The fact that the current peak occurs in advance of the AP may be partially explained by a phase delay between the transmembrane current and potential. The failure of the average current trajectory to return to control values immediately following the peak argues for a positive role of the declining phase in spike triggering. 7. Probability densities preceding spikes were Gaussian, indicating that the average was also the most probable value. Although the densities were broad, confirming that spikes were preceded by a wide variety of current wave forms, their standard deviations were reduced significantly with respect to controls, suggesting preferred status of the average current trajectory in spike triggering. 8. The matrix of correlation coefficients between current pairs suggested that spikes tended to be preceded by wave forms that in part kept close to the average current trajectory and in part preserved its shape. 9. The average first and second derivatives of spike-evoking epochs revealed that current slope and acceleration, respectively, were most crucial in the last 200 msec before spike triggering, and that these dynamic stimulus components were more important for a cell maintained under a depolarizing, rather than a hyperpolarizing bias. 10...


Changes in electrical properties of rat myometrium during gestation and following hormonal treatments.

1. The membrane properties of the rat myometrium, during gestation and following ovarian hormone treatment, have been investigated with the micro-electrode technique. 2. Spontaneously generated bursts of electrical activity alternating with silent periods were recorded from non-pregnant, pregnant and post-partum myometria. The membrane potential was highest during the middle stage of gestation, but the spike amplitude within a burst was not uniform. In the final stage of gestation and during parturition, the membrane potential was low and the spikes within a burst were of low frequency and uniform amplitude. 3. During parturition and post-partum, a gradual depolarization of the membrane, accompanied by an increase in membrane resistance, occurred before the generation of a burst. 4. Excitability of the membrane fluctuated from a peak just before the generation of a burst to a low after the cessation of a burst. 5. Displacement of the membrane potential by electrical current or by lowering the temperature modified the slope spontaneous depolarization, but the fluctuations of excitability persisted. The Q10 value for the frequency of spontaneous bursts, measured between 36 and 30 degrees C, was 3-8. 6. Hyperpolarization of the membrane increased the maximum rate of rise of the spike, but beyond -70 mV, the rate of rise was reduced. Half-inactivation of spike generation of spike generation occurred at a membrane potential less negative than the interburst potential, indicating that the current carrying system was not fully activated during parturition. 7. In both normal and spayed rats, oestradiol hyperpolarized the membrane and the burst of spikes was generated hyperpolarized the membrane and the burst of spikes was generated on a sustained depolarization. Progesterone slightly hyperpolarized the membrane and burst discharges occurred without a sustained depolarization. Simultaneous treatment with progesterone and oestradiol produced a plateau potential of long duration during burst discharges. 8. The thickness of the muscle layer, length constant of the tissue and time constant of the membrane were measured during gestation and from spayed rats under various hormonal conditions. The length constant of the tissue was increased by oestradiol and was further increased by simultaneous treatment withoestradiol and progesterone. The increase in tissue thickness appeared to have the most marked influence on the length constant. 9. The resting and active membrane properties of the progresterone treated myometrium were similar to those observed during the middle stages of gestation. The oestradiol-treated myometrium did not resemble that during the last stages of gestation and parturition, which was simulated by combination of the two hormones, oestradiol preceding progesterone.


Effects of prostaglandin E2 and oxytocin on the electrical activity of hormone-treated and pregnant rat myometria.

The effects of prostaglandin E2 (PGE2) and oxytocin on the electrical activity of rat myometrium at various stages of gestation and following hormonal treatment have been investigated. 1. PGE2 and oxytocin produce an excitation of the myometrial membrane under all experimental conditions. The sensitivity of the myometrium markedly increases during the last stage of gestation, and at parturition is more than one thousand times greater than in the mid-pregnant rat. The sensitivity of the myometrium to oxytocin increases rapidly during the last stage of gestation but to PGE2 the increase is gradual, beginning in the late middle stage of gestation. 2. During the early middle stage of gestation, the sensitivity of the myometrium to PGE2 and oxytocin is lower than in the non-pregnant myometrium. 3. After oestradiol treatment, the sensitivity to PGE2 and oxytocin increases but the sensitivity is much weaker than that during the parturition. On the other hand, after progesterone treatment, the sensitivity is reduced below that of the castrated rat. 4. Differences in the sensitivity of progesterone-treated and oestradiol-treated myometria to PGE2 and oxytocin are compared to those of the pregnant and post-partum myometrium. The results show that the sensitivity of the myometrium to PGE2 and oxytocin during the early and early-middle stages of gestation can be simulated by progesterone treatment, but that the sensitivity during the last stage of gestation and during parturition cannot be simulated by oestradiol and progesterone treatment.


The connexion between the ion-binding sites of the sodium pump.

1. A study has been made of K influx into human red blood cells in order to determine the influence of internal Na on the affinity of the Na pump for external K. Cells were prepared to contain minimal K and two Na concentrations (about 10 and 30 mueqiv/ml. cells) and incubated in solutions with a range of K concentrations. 2. In choline (Na-free) Ringer, activation of K influx by external K was hyperbolic. The Km for external K increased as the internal Na concentration was raised. The increase was greater than the increase in K influx with saturating external K. The Km for external K increased towards a limiting value as the internal Na concentration was raised. 3. In contrast, in Na-Ringer, the activation by external K was sigmoidal and the affinity for external K was independent of the internal Na concentration. 4. K influx was measured at two submaximal levels of external K with a range of internal Na. The affinity for internal Na fell as external K was raised. 5. The results suggest that in the absence of competition between Na and K on the same side of the membrane, there is a connexion between the ion-binding sites of the Na pump.


Joint sense, muscle sense, and their combination as position sense, measured at the distal interphalangeal joint of the middle finger.

1. An anatomical peculiarity allows the hand to be positioned so that the terminal phalanx of the middle finger cannot be moved by voluntary effort. When positioned in this way only joint and cutaneous mechanisms subserve position sense. By altering the position of the hand the muscles are again engaged and able to move the finger. Moving the joint then also excites muscular afferents. 2. The position sense of twelve subjects was assessed with and without engagement of the muscles at the joint. Three tests were used in which either angular displacement, angular velocity or duration of displacement were held constant. 3. When muscular attachment was restored, performance in all tests was greatly enhanced. As engagement of the muscles caused little change in the 'stiffness' of the joint, it is unlikely that the improved performance resulted from increased discharges from the joint receptors. Cutaneous mechanisms are unlikely to mediate this improvement as they are likely to have been unaffected by engagement of muscles. It is concluded that intramuscular receptors are partly responsible for normal position sense. 4. In seven of the twelve subjects the test finger was anaesthetized to isolate the contribution of intramuscular receptors. This muscle sense was variable. In some subjects it provided accurate kinaesthetic information but in others the information was crude. If with the test finger anaesthetized subjects exerted voluntary tension with the muscles that move the joint the muscle sense was improved.


Lung inflation and alveolar permeability to non-electrolytes in the adult sheep in vivo.

1. Experiments were performed on adult sheep to determine the effect of lung distension on the passive permeability of alveoli to water soluble non-electrolytes. With the animal breathing oxygen spontaneously, a segment of one lung was isolated by passing a balloon-tipped catheter through a tracheostomy into a distal bronchus. This isolated atelectatic segment was filled with an isosomotic saline solution containing radio-labelled solutes of known molecular size: [125I]albumin, [14C]inulin, [14C]-sucrose, [3H]mannitol, and [14C]urea. The segment was inflated with oxygen either to a preselected pressure, or to a predetermined fraction of its capacity. Inflation was then maintained for several 10-15 min periods between which the oxygen supply was disconnected and the saline sampled, allowing the tracer concentrations to be measured. 2. At low inflating pressures (20-32 cmH2O) and at low volumes (24-54% of capacity), alveolar permeability to water soluble solutes was slight and could be characterized in terms of a membrane penetrated by cylindrical water filled pores of 0-5-1-6 nm radius. In all experiments showing restricted diffusion, absorption of saline occurred. 3. There was a positive correlation between the degree of lung inflation and pore radius in both the pressure controlled and volume controlled experiments. At high inflating pressures and at inflation volumes which were close to the total capacity of the isolated segment, restriction of solute diffusion was lost; in five out of six such experiments there was a net movement of liquid into the alveoli. 4. These results can be explained by postulating that as the lung epithelium is progressively stretched there is an opening up of water filled channels between alveolar cells. At peak inflation, restriction of diffusion of water soluble solutes is lost, and the alveolar epithelium ceases to function as a barrier between the circulation and air spaces.


Inhibition and facilitation of antidromically identified tubero-infundibular neurones following stimulation of the median eminence in the rat.

1. Stimulation of the median eminence of female rats inhibited the spontaneous firing of antidromically identified tubero-infundibular units. Some units could be inhibited by stimuli subthreshold for the antidromic spike. A conditioning stimulus of subthreshold intensity for the antidromic spike also delayed or abolished the invasion of the soma and dendrites of the same unit by antidromic action potentials evoked by a suprathreshold stimulus given 2-10 msec later. 2. Although strychnine (0-2 mg/kg, i.v.) did not significantly alter the inhibition evoked by stimulation of the median eminence, it was abolished by picrotoxin (2-6 mg/kg, iv.). 3. In seven of the fifty-seven identified units examined stimulation of the median eminence facilitated the spontaneous firing. However, after an i.v. injection of picrotoxin the facilitory response was observed in thirty-seven of the forty-six units tested. Post-stimulus time histograms obtained from the same unit before and after an injection of picrotoxin demonstrated that the latency and duration of the facilitation did not always coincide with that of the inhibition. 4. After an injection of picrotoxin the field potential evoked by antidromic stimulation of the median eminence consisted of a small positive wave followed by a negative wave. Frequently the negative wave of the field potential was accompanied by a convulsive discharge. The latency of the negative wave appears to be identical to that of the facilitation seen in nearby single units. 5. The facilitation evoked by antidromic stimulation in the presence of picrotoxin was blocked by an i.v. injection of alpha-methyl-p-tyrosine (250 or 375 mg/kg). None of the nine units sampled from rats pre-treated with alpha-methyl-p-tyrosine injected twice I.P. (250 mg/kg for each) were facilitated by stimulation of the median eminence following I.V. picrotoxin, while eight of the eleven units sampled from control rats pre-treated with L-tyrosine could be facilitated by antidromic stimulation. 6. These results suggest that tuber-infundibular neurosecretory neurones are inhibited and facilitated by neural pathways which could involve the axon collaterals of the neruosecretory neurones which project to the external layer of the median eminence. It is also suggested that GABA-releasing neurones mediate the inhibition and catecholaminergic neurones are involved in the facilitory pathways. Presumably the facilitation is normally masked by the activity of the GABA-releasing neurones.


The interaction between cutaneous and spinal therman inputs in the control of oxygen consumption in the rat.

1. The effect of thermal stimulation of the spinal cord on the rate of oxygen consumption was studied in five unanaesthetized rats during exposure to various ambient temperatures. 2. In a warm environment, cooling the spinal cord had no effect on the rate of oxygen consumption but in thermoneutral and cold environments the rate of oxygen consumption increased proportionally to the intensity of spinal cooling. Heating the spinal cord decreased the level of oxygen consumption and, if intense enough, suppressed the thermoregulatory increase in metabolic rate. 3. It is concluded that, in the control of oxygen consumption in the rat, the afferent signals from thermal sensors in the spinal cord and skin are added.


Possible presynaptic inhibition in rat olfactory cortex.

1. Field potentials were evoked from rat olfactory cortex slices in vitro at room temperature by lateral olfactory tract stimulation. At very slow stimulation rates (less than 0-1 Hz) a delayed negative wave (late N-wave) was found to follow the first negative wave (N-wave). This late N-wave started 30 msec after the stimulus and lasted for 50-150 msec. 2. Low Ca2+/high Mg2+ medium abolished the late N-wave more rapidly than the N-wave, suggesting a possible multisynaptic origin. 3. The GABA antagonist bicuculline (10(-6) M) abolished the late N-wave without affecting the N-wave. 4. During the late N-wave, both the tract action potential and the N-wave to a second stimulus were reduced. This was attributed, at least in part, to collision with antidromic action potentials which could be detected during the late N-wave. 5. These findings are discussed in terms of a possible presynaptic depolarizing action of a GABA-like transmitter giving rise to presynaptic inhibition and the late N-wave.


Conduction velocity along the afferent vagal dendrites: a new type of fibre.

1. We systematically calculated the conduction velocity along the peripheral extensions of sensory vagal neurones in cats (the dendrites). In addition, a study of excitability cycle and light microscopic investigation were also conducted on these neurones. 2. The conduction velocity of the three known types of fibres (A, B and C) remains uniform along the dendrites. 3. Another mixed type of fibres exists with a C conduction velocity (mean value 1-5 m/sec) along its distal pathway and a B conduction velocity (mean value 6 m/sec) along its proximal pathway. The change in conduction velocity progressively occurs in the thoraco-cervical portion of the vagus nerve at least 20 mm from the receptor and at least 40 mm from the T cell. 4. The mixed fibres exhibited a C type excitability cycle in their peripheral pathway and a B type excitability cycle in their central pathway. 5. The histological study using the teasing method demonstrated the existence of unmyelinated fibres, in the thoraco-cervical region of the vagus nerve, becoming progressively myelinated from the periphery to the nodose ganglion. These fibres are likely to be the ones showing mixed electrophysiological properties. They represent (approximately) 10% of the vagal nerve population. 6. We propose to call the mixed fibres BC because they present electrophysiological and morphological properties of C fibres in their distal part and properties of B fibres in their proximal part.


Increment sensitivity in humans with abnormal visual experience.

1. Visual acuity is lower for gratings oriented diagonally than for those of horizontal and vertical orientations. In addition to this oblique effect, some subjects show substantial deficits in acuity for horizontal or vertical targets (meridional amblyopia). These subjects are invariably astigmatic, but the condition has a neuronal basis and is thought to arise from faulty post-natal neural development. 2. Foveal increment sensitivites have been determined for normal subjects and meridional amblyopes using bar-shaped targets of various lengths, widths and orientations.3. Normal subjects do not exhibit differences in sensitivity as a function of orientation. No oblique effect is found for 1-5' wide bars ranging in length from 10 to 60'. On the other hand, meridional amblyopes have substantial differences in increment sensitivity which depend on test target orientation. Invariably, when there is a deficit in acuity for a particular grating orientation, there is also a reduction in increment sensitivity for a bar of the same orientation. This effect is diminshed or eliminated when the background illuminance is lowered from 70 to 7td. 4. The orientational differences in increment sensitivity found in meridional amblyopes do not increases for bars longer than about 10'. As the bar is shortened, the differences are reduced, and they are absent when the test bar is 6' or less. 5. In normal subjects, for a 1 degree long bar, increment sensitivity increases with width up to about 4' where the width-sensitivity curve levels off. No orientation differences are exhibited. Prominent orientation differences are found with meridional amblyopes when the bar target width is altered. The normal meridian is similar to those of the control subjects but the deficity meridian has very low sensitivity and summation is present for widths up to about 11'.


Identification of pathways mediating cardiovascular responses elicited by stimulation of the septum in the rat.

1. Experiments were done in rats anaesthetized with sodium pentobarbitone to localize pathways mediating the cardiovascular responses elicited by electrical stimulation of the septum. The major efferent projections from the septum were first identified anatomically by the Fink-Heimer II technique and these pathways were subsequently lesioned in acute experiments to establish their role in the mediation of the cardiovascular responses elicited by septal stimulation. 2. Electrical stimulation of histologically localized sites in the lateral septum elicited hypotension and bradycardia whereas stimulation of sites in the medial septum elicited hypertension and bradycardia. 3. Selective lesions of cardiovascular responsive sites in either the lateral or medial septum produced a pattern of degeneration essentially similar to that of previous anatomical studies, i.e. the main efferent projections were localized to the fornix, stria medullaris and the medial forebrain bundle. 4. Stimulation of the fornix did not elicit cardiovascular changes and lesions of this pathway did not alter cardiovascular responses to septal stimulation. 5. Stimulation of the stria medullaris elicited hypotension and bradycardia whereas stimulation of the medial forebrain bundle elicited hypertension and bradycardia. Ipsilateral lesions of the stria medullaris, signicicantly attenuated the hypotension and bradycardia elicited by stimulation of the lateral septum but did not affect the responses to stimulation of the medial septum. Bilateral lesions of the medial forebrain bundle in the region of the lateral hypothalamus abolished the hypertension but did not affect the bradycardia elicited by medial septum stimulation. Cardiovascular, reponses elicited by stimulation of the lateral septum were not affected by medial forebrain bundle lesions. 6. It is suggested that the cardiovascular responses elicited by stimulation of the lateral septum are mediated via the stria medullaris and that the hypertension elicited by stimulation of the medial septum is mediated via the medial forebrain bundle. On the other hand, the bradycardia elicited by stimulation of the medial septum is probably mediated by a pathway presently unknown.


Lithium protection against oxygen toxicity in rats: ammonia and amino acid metabolism.

1. The use of Li pre-treatment in rats before high pressure oxygen exposure has been reported effective in controlling convulsions. This is an effect which is better demonstrated if exposure to oxygen follows shortly after Li injection than exposure following several hours later. 2. This study has investigated the hypothesis that the protective action of Li may be exerted, in the short term, by its removing ammonia from the blood and alleviating the latter's known toxic action. 3. A normal Li distribution time profile in unstressed rat brain and blood following intraperitoneal injection has been established. Brain and blood ammonia, amino acids and Li concentrations were also measured in Li-treated animals exposed and convulsed by oxygen. These measurements were made both shortly (15 min) and also several hours after (24 hr) Li treatment. Ammonia and amino acid values in Li-protected groups were compared to normal unstressed animal values and also to values in animals convulsed by oxygen unprotected by Li pre-treatment. 4. In rat brain abd blood significant (P less than 0-001) elevation of ammonia and glutamine and depression of gamma-amino butyric acid (brain only) and glutamate was noted following oxygen treatment in unprotected animals. Prior injection of Li 15 min before high pressure oxygen exposure delayed convulsions twice as long. Additionally if these animals were only exposed to oxygen for a period of time equal to that which would normally produce convulsions in unprotected animals, brain and blood ammonia and amino acids were maintained near to unstressed animal levels. Concomitantly, blood Li concentrations were considerably depressed below the values one would expect from the previously determined Li distribution time profile. 5. In rats exposed to high pressure oxygen 24 hr after Li treatment there was no protective action against high pressure oxygen convulsion, rather a potentiating effect for convulsion was seen. 6. These data present compelling evidence for the controlling effect of Li in rats, on rising blood ammonia concentration which occurs in high pressure oxygen exposure. The effect might well be due to the known chelating properties of Li with ammonia.


Sulphate excretion by dogs following ingestion of ammonium sulphate or meat.

1. When normal conscious dogs were given small doses of (NH4)2SO4 by stomach tube, the rate of excretion of SO4 increased due to increase in plasma SO4 without change in creatinine clearance. After eating meat, increased SO4 excretion was due to increase in glomerular filtration rate without change in plasma SO4. 2. The calculated rate of tubular reabsorption of SO4 did not change significantly either after (NH4)2SO4 or after meat. 3. The rate of production of SO4 after meat was estimated. In 4 hr approximately one fifth of the S in meat was converted to inorganic SO4; in the same time approximately one third of the N in the meat was converted into urea. 4. Half of the SO4 in the doses of (NH4)2SO4 was absorbed from the gut; all of the NH was converted into urea. (NH4)2SO4 thus produced acidosis. 5. After (NH4)2SO4, increased excretion of NH4+ accompanied the increased excretion of SO4-but Na+, K+ in urine also increased.


Progressive enhancement in the secretory functions of the digestive system of the rat in the course of cold acclimation.

1. The secretory function of the exocrine pancreas and the stomach have been  studied in the course of cold acclimation of rats that had been fed at an ambient temperature of 1 degree C in a climatic room. 2. The secretory responses of pancreatic enzymes evoked by continuous infusion of pancreozymin (PZ, 2-5 mu./kg. hr) and a rapid single injection of PZ (1.7 mu./kg) reached a maximum in the group of rats fed at 1 degree C for 4  weeks, and fell to the control levels after 8 weeks. The increase in the flow of pancreatic juice evoked by single injection of PZ was maximal at 4 weeks and slightly decreased after 8 weeks. 3. The insulin (3-0 i.u./kg) evoked secretion of pancreatic enzymes gradually increased after cold exposure, reached a maximum at 4 weeks and fell to the control levels after 8 weeks. The flow of pancreatic juice after insulin injection was almost the same in every group throughout the course of cold exposure. 4. The ratio of amylase to the total amount of the protein in the pancreatic juice decreased abruptly, in contrast to an increase in the ratio of protease in the process of cold acclimation. The change in the ratio of enzyme activity in the pancreatic juice may reflect parallel changes in enzyme activity in the exocrine pancreas. 5. The gastric secretion in response to insulin and bile secretion in the group fed at 1 degree C for 7 weeks was significantly higher than that in the control group. 6. It was thus concluded that the secretory activities of digestive system were enhanced by prolonged cold exposure and then returned to control level, and that the activites of the pancreatic enzymes were altered in the process of cold acclimation in rats.


Sodium-dependent influx of orthophosphate in mammalian non-myelinated nerve.

1. The rate of uptake of radiophosphate was measured in desheathed vagus nerves of rabbits mounted in an apparatus where the incubating solution flowed along the preparation. 2. Replacement of the Na of the Locke by either choline or Tris slowed the rate of uptake to about 10% of its value in Na; with K it was slowed to 20%; and with Li to 50%. 3. Measurements of the rate of uptake at different extracellular phosphate concentrations showed that in choline-Locke the influx of phosphate was proportional to the extracellular phosphate concentration, while in Locke the rate of inward flow showed a tendency to saturation with increasing phosphate concentrations. 4. Extracts of nerves after 45 min exposure to labelled solutions showed for different phosphate concentrations a slowing of the labelling of ATP, ADP and creatine-phosphate (CrP) and inorganic phosphate (Pi) when Locke was replaced by choline-Locke. 5. A similar slowing was found when, at 0-2 mM phosphate, the labelling of these compounds was measured at different times: application of choline-Locke reduced the rate of incorporation of 32P to about 15% of the rate in Locke. 6. In preparations that were loaded with radiophosphate and then washed with inactive solution, and the effluent fractionated, over 90% of the radioactivity was found in the inorganic phosphate fraction. 7. The efflux of 32P showed an initial exponential phase with a time constant of 20-30 min and a much slower one. The slow efflux had a rate constant of 0-0014 min-1 in 0-2 mM external phosphate. 8. Increasing the external phosphate concentration increased the efflux. 9. Prolonged incubation in choline-Locke reduced the efflux. 10. The influx of phosphate was calculated for different external phosphate concentrations using the rate of uptake of radiophosphate measured 45 min after the application of isotope and the corresponding efflux rate coefficient and the intracellular specific activities of the labelled compounds. 11. A correction was also introduced for diffusion, using the "limited biophase model". 12. The Na-sensitive influx, i.e. the difference between influx in Locke and influx in choline-Locke, showed saturation kinetics. 13. A Lineweaver-Burk plot for Na-sensitive uncorrected influxes gave a vmax of 16-7 mumole/kg wet wt. min and an apparent Km of 0-42 mM; for the corrected fluxes these values were 18-2 and 0-36. 14. It is concluded that a large part of phosphate influx and some of the phosphate efflux is mediated by a specific Na-dependent phosphate transport system. 15. This system seems to be present also in other types of nervous tissues and probably in many types of animal cells.


The morphology of spinocervical tract neurones in the cat.

1. The morphology of physiologically identified spinocervical tract (SCT) neurones was studied using the intracellular injection of Procion dyes in anesthetized and decerebrate cats. 2. Extracellular recordings were made from SCT neurones at depths between 1000 and 2850 mum from the cord surface but neurones were only stained at depths between 1100 and 2400 mum. 3. The dendritic trees of stained SCT neurones were reconstructed in the transverse plane of the spinal cord. All SCT neurones had well developed dorsal dendrites but despite this it is not possible to consider the twenty-two SCT cells in out sample as consituting a morphologically homogenous population. 4. There was no correlation  between the form of the dendritic trees and the depth of SCT neurones in the dorsal horn as determined both from measurements from the dorsal grey-white border and the position of cells with respect to the border between Rexed's laminae II and III. 5. Six types of SCT neurones were identified on the basis of the form of their dendritic trees as viewed in the transverse plane: (1) radially symmetrical, (2) semicircular, (3) large elliptical, (4) bilobed, (5) triangular, (6) small elliptical. Each of these types was found only in a certain region across the dorsal horn although any one region could contain more than one type. 6. Spinocervical tract neurones with small elliptical dendritic trees always had receptive fields encompassing part of the hip or thigh and were unique in being located in the lateral portions of the horn. 7. There was no correlation between the morphology of SCT neurones and their excitatory cutaneous inputs, receptive field size, axonal conduction velocity or depth in the dorsal horn.


The secretion of citrate into milk.

1. The time course of changes in specific activities of citrate, lactose and fatty acids in milk during frequent milking, following the I.V. administration of labelled glucose, acetate and chylomicrons in goats has been studied. Peak specific activities of lactose and citrate in milk were reached at 2-3 hr, while peak specific activites of fatty acids were reached at 5-7 hr. 2. Following short I.A. infusions of 24Na, 36Cl, and 42K, peak specific activities in milk were reached in 1 hr or less. 3. The mammary epithelium of lactating goats was found to be virtually impermeable to labelled citrate in both directions. 4. Labelled citrate had an apparent volume of distribution in lactating guinea-pigs mammary slices in vitro similar to that of extracellular space markers. 5. Treatment of goats with large doses of oxytocin markedly increased the permeability of the secretory epithelium to labelled citrate. 6. In the goat mammary gland, citrate, protein and calcium failed to enter milk which had been diluted with isosmotic lactose by intraductal injection, whereas Na, K and Cl did enter, thus tending to restore the concentrations of these ions to normal. 7. It is suggested that citrate, which is formed within the sucretory cell, enters milk not by passage across the apical cell membrane but, in common with lactose and milk protein, by exocytosis of Golgi vesicles. It appears that citrate is held at high concentrations in milk by virtue of the impermeability of the mammary epithelium to the forms in which it occurs in milk.


Release of noradrenaline from the cat spleen by nerve stimulation and potassium.

1. Release of noradrenaline from the perfused cat spleen, or from isolated spleen slices, in response to prolonged nerve stimulation or maintained depolarization by potassium was measured. 2. Prolonged stimulation of the splenic nerves at 2, 10 and 30 Hz for 10 min evoked release, which was maximum during the first 2 min, and then declined during the remaining period of stimulation. When noradrenaline release was induced by high potassium from the perfused spleen or from isolated slices, it followed a similar time course to nerve stimulation. Similar results were obtained from phenoxybenzamine-treated spleens, using both modes of stimulation. 3. Stimulation of the splenic nerves in calcium-free Krebs solution did not release noradrenaline. If calcium was introduced at a later stage during stimulation, the release was markedly diminished. In phenoxybenzamine- or phentolamine-treated spleens, stimulation of the nerves in the presence of calcium evoked a secretory response which was comparable to the one produced by introduction of calcium after a few minutes of nerve stimulation. 4. Simultaneous application of calcium plus high potassium always produced a much greater secretion of noradrenaline than application of calcium after a few minutes of potassium depolarization. Release of noradrenaline by potassium from phenoxybenzamine-treated spleens was also much greater if calcium and potassium were added simultaneously than addition of calcium after a few minutes of potassium depolarization. 5. In the presence of maintained depolarization by potasssium, tyramine was effective in causing release of noradrenaline.


The secretory characteristics of dehydrocholate in the dog: comparison with the natural bile salts.

1. During dehydrocholate administration in the taurine replete dog, the maximum excretory rate of total bile salt (almost entirely dehydrocholate derivative, mostly conjugated) was 3-84 +/- 0-53 (S.D.) mumole/min. kg body wt. (eleven experiments). This was much less than the excretory maximum previously obtained for taurocholate (8-64 +/- 1-31 (S.D.) mumole/min. kg total cholate, mostly conjugated). 2. The superimposition of taurocholate infusion did not cause any significant change in the 'dehydrocholate' maximum but taurocholate itself was excreted into bile at no more than about half its normal maximum. When taurocholate maximum excretion was established first, it was reduced by dehydrocholate administration. In both types of experiment the joint bile salt excretory maximum was of the same order as that of taurocholate alone, provided taurocholate made up at least 40-50% of the total bile salt. 3. When taurocholate administration was stopped, the maximum excretory rate of 'dehydrocholate' rose to values up to 63% above the initially determined excretory maximum; the enhanced 'dehydrocholate' excretory maximum, when calculated for optimal conditions, approached that of actively conjugated vholate, even though the effective 'dehydrocholate' concentration in bile was ten to twenty times the critical micellar concentration of taurocholate. This suggests that the effective bile salt concentration in bile is not an important determinant of the secretory performance of a bile salt. 4. To explain findings (2) and (3) it is necessary to postulate that taurocholate has both a facilitatory and an inhibitory action on 'dehydrocholate' excretion. The facilitatory action, which persists after taurocholate has left the animal, may consist either of an increase in the maximum rate at which modification of dehydrocholate takes place within the liver cell, or an increase in the number of functioning 'carriers' for 'dehydrocholate' transfer. The data suggest that the inhibitory effect is due to the competitive interaction that also appears to exist between the two bile salts. 5. The increase in bile flow rate per unit increase in 'dehydrocholate' excretion (15 ml./m-mole) was about twice that obtained for taurocholate. There was no significant formation of micellar aggregates during 'dehydrocholate' excretion, as judged from the total electrolyte concentration of bile and its osmalality. 6. During the excretion of 'dehydrocholate'-taurocholate mixtures (approximately 1:1) at submaximal rates the associated bile flow rate was not less than the sum of the separate components, thus suggesting that 'dehydrocholate' was not being incorporated in taurocholate mixed micelles.


The direct effect on pulmonary stretch receptor discharge produced by changing lung carbon dioxide concentration in dogs on cardiopulmonary bypass and its action on breathing.

1. Single fibre pulmonary stretch receptor discharge was recorded in dogs on cardiopulmonary bypass. 2. Inhalation of CO2 depressed pulmonary stretch receptor discharge despite the absence of changes in arterial PCO2. This effect was particularly marked with airway CO2 levels below 5%. 3. Changing arterial PCO2, without changing airway CO2, had only small and insignificant effects on pulmonary stretch receptor discharge. 4. The effect of changes in airway CO2 on pulmonary stretch receptor discharge was rapid and correlated well in time with the reflex tachypnoea produced when CO2 was inhaled in conditions of cardiopulmonary bypass. 5. Stimulation of the central end of the cut vagus nerve was triggered from simultaneously recorded action potentials from a single pulmonary stretch receptor. 6. In these conditions, the reflex response to CO2 could be simulated provided that the pulmonary stretch receptor had an end-expiratory discharge. 7. It is suggested that the vagally mediated tachypnoeic response to changes in airway CO2 seen in conditions of cardiopulmonary bypass is due to the effect of CO2 on the end-expiratory discharge of pulmonary stretch receptors.


Respiratory and thermoregulatory responses of rabbits breathing carbon dioxide during heat exposure.

1. Rabbits were clipped and exposed in turn to three environmental conditions: control (C), cold exposure (CE) and water deprivation (WD). Following each type of treatment, the rabbits were exposed to an ambient temperature (Ta) of 35 degrees C for 1 hr. Throughout this period they breathed either normal atmospheric air or 6% CO2 in air. 2. During heat exposure, measurements were made of the respiratory responses and of the O2 consumption (Vo2) of the rabbits. Rectal temperature (Tre) was measured immediately before and again immediately after heat exposure. 3. When subjected to cold exposure or water deprivation the rabbits showed an initial decrease in respiratory frequency (RF) and an initial increase in VT when compared with controls. There was no difference in VE. Rabbits breathing 6% CO2 showed an increase in VT and VE and a decrease in RF when compared with rabbits breathing atmospheric air. In all cases a change in VT or RF was associated with a reciprocal change in the other parameter. 4. The respiratory responses to breathing 6% CO2 were essentially similar in treated and control rabbits, from which it is concluded that neither cold exposure nor water deprivation alter the sensitivity of the medullary respiratory centre to the respiratory drive from the central chemosensors. 5. The increase in Tre during heat exposure was significantly less in rabbits breathing 6% CO2 than in rabbits breathing atmospheric air. However, there was no significant over-all difference in VO2 between rabbits breathing CO2 and those breathing air. From this it is concluded that increased ventilation induced by CO2 causes a greater dissipation of heat than does thermally-induced panting. 6. It is concluded that VT is controlled by the level of blood PCO2 whereas RF is controlled by thermoregulatory requirements. It is further concluded that the reciprocal relationship between VT and RF is regulated in such a way as to maintain VE at the appropriate level for effecting gaseous exchange and evaporative heat loss.


Polyneuronal innervation of skeletal muscle in new-born rats and its elimination during maturation.

1. The events taking place during the elimination of polyneuronal innervation in the soleus muscle of new-born rats have been studied using a combination of electrophysiological and anatomical techniques. 2. Each immature muscle fibre is supplied by two or more motor axons which converge on to a single end-plate. There was no sign of electrical coupling between muscle fibres receiving multiple synaptic inputs. By the end of the second week after birth virtually all muscle fibres are innervated by only a single motor axon. 3. The average tension produced by individual motor units, measured in terms of the percentage of the total muscle twitch tension, declined dramatically during the first 2 weeks after birth. During this period there was no significant change in the number of motor neurones innervating the soleus muscle. Thus, the disappearance of polyneuronal innervation reflects a decrease in the number of peripheral synapses made by each motor neurone. 4. The decline in motor unit size was delayed, but not ultimately prevented, by the early surgical removal of all but a few motor axons to the soleus muscle. This procedure also caused a delay in the removal of polyneuronal innervation involving the remaining motor units. 5. Following a crush of the soleus nerve in neonatal animals, regenerating axons usually returned to the original end-plates. Polyneuronal innervation was extensive at early stages of re-innervation and it disappeared during the second week after birth just as in normal muscles. 6. Cross-innervation of neonatal muscles by an implanted foreign nerve caused a rapid disappearance of cholinesterase at denervated original end-plates and in most fibres prevented re-innervation by the original nerve. In the small proportion of fibres that did become innervated through both the foreign and original nerves the end-plates were more than 1 mm apart, and both foreign and original nerve end-plates could persist indefinitely. 7. Many cross-innervated fibres received multiple inputs through the foreign nerve. Some foreign end-plates were separated by distances ranging up to 1 mm. Polyneuronal innervation through the foreign nerve was completely eliminated during maturation but over a slightly longer period than in normal muscles. Apparently the elimination process can act over a distance up to but not much more than 1 mm. 8. These observations suggest that there are several factors influencing the elimination of redundant inputs in immature muscles. Individual motor neurones appear to have an inherent tendency to withdraw the majority of their original complement of peripheral terminals. The determination of which particular synapses are to survive, however, seems to be made in the periphery by a selection among all the synapses that innervate a limited region of each muscle fibre. There may be a competitive interaction among synapses in which those belonging to smaller motor units are less likely to be eliminated, thereby leading to a relatively uniform size of the motor units in the soleus.


The conditions required for the maintenance of binocularity in the kitten's visual cortex.

1. In young kittens, cortical neurones, which are usually binocularly driven, have their binocularity reduced if one eye is covered, or if the eyes are made strabismic or alternately occluded. Some of the factors causing these changes were analysed. 2. If the contrast of one retinal image is abolished with no difference in mean illumination, the input from that eye is virtually lost. 3. If one eye merely has its mean retinal illumination attenuated, that eye does not specifically lose its influence in the cortex, although there is a reduction in the proportion of binocular units. This change might partly be due to a difference in the timing of signals from the two eyes but is more likely to be caused by a difference in the strength of the discharges. 4. There is little change in binocularity if one image is dimmed but contrast is absent from both. 5. If contours of very different orientation fall simultaneously on corresponding retinal regions, binocularity breaks down, as in the case of strabismus or when different patterns are presented to the two eyes. But as long as the patterns on corresponding retinal points have similar orientation, even if the visual axes are misaligned, binocularity can be maintained. 6. If the eyes are not stimulated simultaneously, binocularity is reduced, even if the contours falling on the two retinae (at different times) are identical. 7. Roughly simultaneous stimulation, with roughly congruent patterns on the two receptive fields, are needed for the upkeep of binocular connexions on to cortical cells.


The possible mechanism for the 'fade' in acid gastric secretion during continuous infusions of pentagastrin.

1. The 'fade' phenomenon seen in acid gastric secretion in response to a continuous intravenous infusion of pentagastrin has been studied in anaesthetized cats. Whereas acid secretion to a continuous infusion of histamine steadily increased for 3 hr, that to pentagastrin infusion rose steeply to a maximum at 45 min and then decreased gradually to reach about 60% of maximum after 2 hr. 2. Pre-treatment of the cats with the 5-hydroxytryptamine (5-HT) blocking drugs, methysergide and cyproheptadine or with the 5-hydroxytryptamine-depleting drug reserpine completely altered the pattern of secretion to pentagastrin such that the response was similar to that seen during an infusion of histamine. These drugs did not affect secretion stimulated by histamine. 3. It is suggested from these results that pentagastrin releases 5-hydroxytryptamine which at first potentiates and then inhibits the resulting stimulated secretion.


Competitive and non-competitive re-innervation of mammalian sympathetic neurones by native and foreign fibres.

The ability of native (sympathetic preganglionic) and foreign (vagal) nerve fibres to re-innervate neurones of the guinea-pig superior cervical ganglion, either alone or in competition with each other, has been studied by means of intracellular recording and electron microscopy. 1. Native fibres make synaptic contacts with nearly all ganglion cells within one month of cervical trunk section; within 6 months the degree of innervation, judged by measurement of excitatory post-synaptic potential (e.p.s.p.) amplitude and electron microscopical synapse counts, approaches normal. However, even after 15 months innervation was weaker than in normal control ganglia. 2. Vagal fibres are less successful during re-innervation. Although a similar number of foreign fibres grown into denervated ganglia and make contact with nearly all ganglion cells within a month, after 6-12 months e.p.s.p. amplitudes in response to foreign nerve stimulation remain relatively small, and counts of synapses are only about 60% as great as in ganglia re-innervated with the native nerve. 3. When both native and foreign fibres are allowed to re-innervate ganglion cells simultaneously, about half the neurones in the ganglion receive synapses from both sources after 1 month. The proportion of dually invervated cells remains roughly constant for at least 14 months. Neither set of preganglionic fibres dominates or displaces the other, although neurones generally are re-innervated more effectively by native than foreign fibres, as is true during non-competitive re-innervation. 4. Thus during re-innervation of mammalian sympathetic neurones native fibres are preferred to foreign ones only in the sense that roughly the same number of native fibres form many more synapses on ganglion cells than do vagal axons. A foreign synapse, once formed, is as stable as a native one, and shows no tendency to be replaced by native terminals. These findings are discussed in relation to other evidence which has suggested specificity and selectivity during re-innervation of mammalian autonomic neurones.


A new method for labelling saxitoxin and its binding to non-myelinated fibres of the rabbit vagus, lobster walking leg, and garfish olfactory nerves.

1. A new method of labelling saxitoxin (STX) is described, based on transfer of tritium from tritiated water to the toxin. 2. The radiochemical purity of the labelled toxin has been directly determined, rather than being based on indirect biochemical means, as in previous experiments with Wilzbach-labelled STX and TTX. 3. The specific activity of the labelled toxin, 66 d..m.f-mole-1, corresponds with one tritium atom per molecule STX, an improvement of about 300-fold over other means of labelling TTX and STX. 4. The binding of this toxin to rabbit, lobster and garfish olfactory nerve fibres has been re-examined. 5. The density of sodium channels calculated on the basis of the binding of the toxin is about four to six-times the values previously reported.


Non-random distribution of cell types in the preoptic and anterior hypothalamic areas.

1. Single units were recorded extracellularly from the preoptic and anterior hypothalamic areas of male and female rats anaesthetized with urethane. 2. The effect of single shock stimulation of both the mediobasal hypothalamus and corticomedial amygdala upon unit activity was assessed for all cells. Neurones were designated either antidromically invaded, orthodromically excited or inhibited, or non-responsive. 3. The distribution of responses among recorded cells was analysed for two groups of neurones. In one group the intercell distance was less than 100 mum and in the second group the distance was more than 1 mm. 4. For widely separated cells the connexions with the corticomedial amygdala and mediobasal hypothalamus were distributed at random. 5. In the groups of cells clustered closely together a marked structuring of connexions was apparant. In particular, there was a significant correlation between orthodromic responses from the amygdala (P less than 0-05) and cells antidromically activated and orthodromically excited by stimulation of the mediobasal hypothalamus were frequently adjacent to one another (P less than 0-02. 6. A complimentary analysis demonstrated that cells in the ventral half of the recording zone received significantly more inhibitory inputs from both the mediobasal hypothalamus (P less than 0-001) and the corticomedial amygdala (P less than 0-02) than those situated more dorsally. 7. The data provide evidence for structuring and orderliness in hypothalamic connexions that is often not apparent from descriptions of electrophysiological experiments.


The potentiating influences of insulin on pancreozymin-induced hyperpolarization and amylase release in the pancreatic acinar cell.

1. Insulin (1 mu./ml.) potentiated the release of amylase from isolated pancreas of rats perfused with erythrocyte-containing medium and stimulated by 0-5 m-u. pancreozymin (Pz)/ml., whereas the same concentration of insulin failed to potentiate the response evoked by or 200 m-u. PZ/ml. 2. Intracellular measurement of membrane potentials from the acinar cells of the same preparations showed that insulin (1 mu./ml.) simultaneously potentiates the hyperpolarization and amylase release in response to 0-5 m-u. PZ/ml. 3. These effects of insulin on the PZ-induced responses were inhibited by ouabain (3 X 10(-5) M). 4. The results suggest that insulin, in the concentration studied, has a potentiating action on the activity of an electrogenic Na pump, which maintains the hyperpolarization and amylase release during continuous stimulation with Pz. 5. Insulin in also potentiated the Pz-induced amylase release in the rat pancreata in situ after vagotomy and ligature of the pyloric region.


Sialotonin: vasopressor substance in saliva and submandibular gland of the cat.

1. The large molecular pressor agent, sialotonin, present in cat saliva, has been separated from kallikrein and characterized further. 2. Sialotonin is present in saliva produced by parasympathetic (ch8da) but not in that produced by sympathetic nerve stimulation. After degenerative preganglionic parasympathetic nerve section, both sialotonin and kallikrein disappear completely, or nearly so. Sympathetic nerve section, however, fails to affect the concentration of these substances in chorda saliva. 3. The sialotonin activity is saliva is unaffected by ligation of Wharton's duct for 3--4 days whereas the kallikrein concentration is greatly reduced. 4. Although normally present in chorda saliva, sialotonin cannot be detected in aqueous extracts of the submandibular gland. It is present, however, in extracts of glands which have previously been subjected to prolonged sympathetic nerve stimulation or to ligation of the duct for 3-4 days. 5. The intravenous or close-aterial injection of sialotonin causes marked but brief reductions of blood flow in the submaxillary gland and intestine.


Calcium and the control of discrete wave latency in the ventral photoreceptor of Limulus.

1. Discrete, transient depolarization (discrete waves) of the ventral photoreceptor of the horseshoe crab, Limulus, occur spontaneously in the dark adapted photoreceptor and are also evoked by light. They form the basic events which comprise the receptor potential. A brief, low energy flash of light evokes variable numbers of discrete waves which have variable latencies. Evidence suggesting that discrete wave latency reflects the kinetics of the chemical reactions of phototransduction is reviewed. 2. The concentration of extracellular Ca influences both the average discrete wave latency and its variability. Lowering extracellular Ca prolongs the latency and increases its variability. Increasing extracellular Ca has the opposite effect. 3. Changes in discrete wave latency caused by changes in extracellular Ca require 10--15 min to become fully manifest, whereas when the concentration of extracellular K is increased the photoreceptor achieves a steady-state depolarization in 10-15 sec. 4. Iontophoresis of the Ca-chelating agent EGTA into the photoreceptor increases both the average discrete wave latency and its variability. Iontophoresis of Ca-EGTA mixtures may either increase or decrease discrete wave latency and its variability depending upon the proportion of Ca mixed with EGTA. 5. It is suggested that the concentration of intracellular rather than extracellular ionized Ca is the prime factor indicating discrete wave latency. The effects of changing extracellular Ca can be explained if the photoreceptor is permeable to Ca in the dark and if it maintains a low intracellular Ca concentration by virtue of active metabolic processes (a pump-leak system). 6. Lowering the temperature of the photoreceptor also has the dual effect of increasing discrete wave latency and its variability. However, effects of lowering temperature and Ca simultaneously are greater than the sum of the two effects in individually. This suggests that Ca may be a reactant in the chemical process of phototransduction. 7. Changing the concentration of extracellular Ca does not change the quantum efficiency of discrete wave production. A previous study showed that quantum efficiency is not changed by temperature. Thus, once initiated by the absorption of light, the reactions that subserve phototransduction may be forced to completion. Ca probably exerts its influence by changing one or more rate constants in the reaction sequence.


The effects of irritation at various levels of the airway upon tracheal mucus secretion in the cat.

1. Sulphated glycoprotein output from the trachea, isolated in situ, has been measured in anaesthetized cats by a radio-isotopic method. The effects of irritation of various parts of the airway on this mucus output were studied. 2. Mechanical stimulation of the nose and nasopharynx increased tracheal mucus output by reflexes which involved parasympathetic and probably also sympathetic motor pathways. 3. Laryngeal stimulation had a similar through the same motor pathways. 4. Inhalation of ammonia vapour into the lower airways reflexly increased mucus output from the isolated trachea. The efferent pathway for this reflex was mainly or entirely parasympathetic. It is argued that the afferent pathway involved cough receptors. 5. Lung inflation, inhalation of histamine aerosol and intravenous injection of phenyl diguanide (which excite mainly lung stretch receptors, lung 'irritant' receptors and alveolar 'J-receptors' respectively) had no consistent effect on tracheal mucus secretion. 6. The afferent and efferent pathways of these reflexes are discussed.


Asymmetric tonic labyrinth reflexes and their interaction with neck reflexes in the decerebrate cat.

1. Tonic labyrinth and neck reflexes were studied separately and in combination in the decerebrate cat with C1 and C2 spinal roots cut. Reflex effects were observed as changes in length of the isotonically loaded medial head of triceps. 2. The tonic labyrinth reflexes acted asymmetrically on the medial head of triceps. Side-down rotation of the head produced shortening in medial triceps, whereas side-up rotations of the head resulted in a lengthening. 3. The tonic neck reflexes acted asymmetrically on the medial head of triceps. Side-down rotations of the neck produced a lengthening of medial triceps, whereas side-up rotations of the neck resulted in shortening. 4. Labyrinth and neck reflexes produce opposite effects on the same limb extensor muscle so that, if the neck innervation is intact, head tilting produces no change in muscle length. 5. It is suggested that the interaction between the labyrinth and neck reflexes contributes to the stability of the trunk, allowing the head to move freely on the body without affecting this stability. Labyrinth and neck reflexes need therefore to be considered together as a single system.


Frequency of tendon organ discharges elicited by the contraction of motor units in cat leg muscles.

1. The responses elicited in individual tendon organs by the contraction of single motor units were studied in peroneus longus, peroneus brevis, tibialis anterior and soleus muscles. 2. No simple relation was found between the discharge frequency of a tendon organ and the tension produced in the muscle tendon by the contraction of individual motor units. 3. The sensitivity of a given tendon organ to contractile tension was not the same for each of the motor units which elicited its discharge. There was no correlation between the sensitivity of the receptor and the strength of the motor units. 4. Upon repetitive stimulation of a tendon-organ-activating motor unit at increasing rates, the frequency of the receptor sustained discharge reached a maximal value for rates of stimulation eliciting submaximal tetanic tension. Higher rates only produced an increase in the dynamic component of the tendon organ response. 5. These observations show that the contractile tension sensed by a tendon organ is not a simple fraction of the tension which appears at the muscle tendon. They might be accounted for as consequences of the fine structure of tendon organs and of variations in the number of muscle fibres contributed by different motor units to the bundle inserted on each receptor. The location of most tendon organs at musculo-aponeurotic junctions rather than in the tendon proper, could also be responsible for some of the observed discrepancies.


Inhibitory effects of acetylcholine on neurones in the feline nucleus reticularis thalami.

1. Short iontophoretic pulses of acetylcholine (ACh) inhibited almost every spontaneously active cell encountered in the nucleus reticularis thalami of cats anaesthetized with a mixture of halothane, nitrous oxide and oxygen. On 200 cells the mean current needed to eject an effective inhibitory dose of ACh was 67 +/- 2 nA. When the ACh-evoked inhibition was mimicked by gamma-aminobutyric acid (GABA) or glycine on the same cell, the current required to release ACh was found to be approximately twice as great as that required to release an equally effective dose of GABA or glycine. 2. ACh inhibitions developed with a latency which was very much shorter than that for ACh excitation in cells of the ventrobasal complex. The latency of the ACh-evoked inhibition was as rapid as the onset and offset of the excitation of the same cells glutamate and their inhibition by GABA or glycine. 3. The firing pattern of ACh-inhibited neurones in the nucleus reticularis was characterized by periods of prolonged, high frequency bursts, and their mean firing frequency was 22 Hz. Raster dot displays and interspike interval histograms showed that whereas ACh suppressed the spikes that occurred between bursts much more readily than those that occurred during bursts, all spikes were equally sensitive to the depressant action of GABA and glycine. Large doses of ACh provoked or exaggerated burst activity. 4. ACh-evoked inhibition was extremely sensitive to blockade by short iontophoretic applications of atropine, which had no effect on the inhibitions evoked on the same cell equipotent doses of GABA or glycine. The ACh-evoked inhibitions were also antagonized by dihydro-beta-erythroidine released with slightly larger currents. When tested on the same cell, small iontophoretic applications of picrotoxin and bicuculline methoiodide blocked the inhibition evoked by GABA but had no effect on that evoked by ACh. Iontophoretic strychnine only rarely affected the inhibition evoked by ACh, while readily blocking the inhibition evoked on the same cell by an equipotent dose of glycine. In two cats, intravenous strychnine (1-2 mg/kg) had no effect on the ACh-evoked inhibition, while greatly reducing the sensitivity of the cell under study to glycine. 5. Only four out of forty-eight ACh-inhibted cells tested were inhibited by iontophoretic applications of either guanosine or adenosine 3':5'-phosphate. 6. Cells of the nucleus reticularis have been shown to have an inhibitory action on the thalamic relay cells, which are excited by ACh. It is suggested that the presence of both ACh excited and inhibited cells in different nuclei of the thalamus could be of considerable functional significance in gating sensory transmission through the thalamus.


The incidence of myocardial infarction.

I studied 183 patients with myocardial infarction including sudden deaths related to ischaemic heart disease occurring in general practice during a period of four years.The figures are examined by age, sex, history of event, and place of treatment. The ratio of male to female infarctions was 2:1, males having a mean age of 63 years and females 71 years.The total death rate for all myocardial infarctions was 46 per cent, but if sudden deaths are excluded, the rate becomes 24 per cent.The annual incidence of myocardial infarction per 1,000 in the practice population of 11,195 was 4.1 and the total annual death rate per 1,000 was 1.9.


The natural history of angina in a general practice.

An appreciation of the natural history of angina pectoris is important when deciding on the place of new and potentially dangerous forms of treatment. During 1950-1975, 268 patients with angina were diagnosed and followed up in my London general practice. The annual incidence, in adults over 40, was five per 1,000 and increased with age.During the period of follow-up, half the patients died, an annual mortality of 4.6 per cent. However, among the survivors one third ceased to suffer anginal symptoms spontaneously and without specific therapy. Of those who continued to suffer from angina, in 71 per cent the condition was graded as minor, in 27 per cent as moderate, and in only two per cent were the attacks severe and disabling. Usually the angina was primary (77 per cent) and it was secondary, after myocardial infarction, in 23 per cent.Of the 134 deaths, three quarters were from a cardiovascular cause. This group of angina patients had a 2:1 times greater observed, than expected, risk of dying (O/E ratio). The O/E mortality ratio fell progressively with age. It was highest in the 40-49 decade (4.0) and lowest in the over 80s, when the observed mortality rate was less than expected (0.9). The O/E mortality ratio was higher in men (2.3) than in women (1.7).From this survey I conclude that angina does not have a uniformly bad prognosis, and that with the advent of beta-adrenergic blockers, the proportion considered for angiocardiography and aorto-coronary bypass grafting should be less than five per cent of all patients with angina.