Lack of a role for cardiac sympathetic nerves in the uptake and metabolism of 3H-5-hydroxytryptamine by isolated rabbit hearts. Perfused rabbit hearts removed 3H-5-hydroxytryptamine (3H-5-HT) from a perfusion solution containing 2.6 ng/ml and a tissue-to-medium ratio of about 10 was achieved after a 55 min perfusion period. 12.4 +/- 1.0% of the cardiac total radioactivity consisted of metabolites and metabolites appeared in the venous effluent during the course of the perfusion. Pretreatment of animals with 6-hydroxydopamine reduced cardiac noradrenaline concentrations by 92%, abolished the histochemical fluorescence characteristic of noradrenaline in sympathetic nerves and abolished the mechanical response to sympathetic nerve stimulation. In these hearts, the removal of 3H-5-HT from the perfusion fluid and the accumulation of total radioactivity was normal. There was, however, a significant increase in the proportion of the total cardiac radioactivity which was present as metabolites. The results suggest that the cardiac sympathetic nerves are not the site of 3H-5-HT uptake when the concentration in the perfusion medium is low. By analogy with work in the lung, it may be the endothelial cells of the coronary microcirculation which are responsible.


Dantrolene effects on neuromuscular function in cat soleus muscle. Dantrolene sodium (DS) was investigated for its effects on cat soleus muscle contractile properties and motor nerve terminal activity in particular. DS, 0.1-1.5 mg/kg i.v., caused a dose-dependent depression of indirectly elicited contractile strength which was more pronounced at lower frequencies of stimulation. Maximum tetanic strength at frequencies of 10-400 Hz was depressed to a lesser degree than contractile responses evoked by lower frequencies of stimulation; the twitch/tetanus contraction ratios were reduced with increasing dose, primarily because of diminished twitch. DS was without effect on motor nerve terminals as evidenced by normal post-tetanic repetition in the nerves following DS administration. Post-tetanic potentiation became relatively larger in amplitude as contractile strength was diminished. These data suggest that DS depresses neuromuscular function at a site other than the neural apparatus at the neuromuscular junction.


Effects of AY-22,241 (Actodigin) on electrical and mechanical activity of cardiac tissues. AY-22,241 (Actodigin) is a new rapid-acting semisynthetic cardiotonic steroid. In experiments on contractility of cat papillary muscle, Actodigin (2 times 10(-7) to 4 times 10(-6) M) produced a dose-dependent positive inotropic effect, a marked increase in the maximum rate of force development, and no change in resting tension. Electrophysiologic studies performed with microelectrode techniques on isolated Purkinje fibers superfused with Tyrode solution, revealed dose-dependent decreases in resting membrane potential, action potential amplitude and duration, and the maximum rate of rise of phase 0 (Vmax). Purkinje fibers superfused with extracorporeally circulated blood from a donor dog receiving 0.075 mg/kg/min Actodigin showed small decreases in resting membrane potential preceding the onset of donor premature ventricular contractions. Progressive decreases in resting membrane potential, action potential amplitude and duration, and Vmax accompanied donor ventricular tachycardia. All effects were rapidly reversible, and compared to ouabain, equi-inotropic concentrations of Actodigin caused significantly less electrophysiologic toxicity.


The effect of atropine on the turnover of acetylcholine in the mouse brain. The effect of atropine on the acetylcholine (ACh) turnover in the mouse brain has been studied and related to the central effect (motor activity) of the drug. At the threshold dose for maximal increase in motor activity, atropine had no measurable effect in the brain on the initial rate of formation of labelled ACh from labelled choline (Ch) i.v. injected. However, if atropine was injected 3 min after the injection of labelled Ch, when the labelled ACh had reached its peak value in the brain, there was a more rapid exponential decline of labelled ACh. This was assumed to be an indication that atropine increases the turnover of ACh in the brain. The specific radioactivity of ACh was not changed 2-17 min after the atropine injection, which indicates that atropine does not preferentially increase the release of newly synthetized ACh.


Comparison of the action of different barbiturates on squid axon membranes. Experiments utilizing squid giant axons have been carried out to determine if the duration of anesthesia obtained from phenobarbital, pentobarbital and hexobarbital can be explained by events related to their action at the nerve membrane level. For comparison, the barbiturates were perfused both outside and inside the squid axon at concentrations which produced an approximately equivalent depression of the maximum rate of rise of the action potential. The time to 50% maximum block and the time to 50% maximum recovery following washing, were compared for all three drugs. All three barbiturates were more potent from inside the squid axon than from the outside. This finding coupled with the fact that the time course of the drug effects was much shorter when applied inside as compared to outside suggests an internal site of action. There was no statistically significant difference between a short acting barbiturate and a long acting barbiturate with respect to their time course of action or recovery when they were both applied to either the inside or outside of the membrane.


The positive inotropic effect of glucagon in the chronically failed right ventricle as demonstrated in the isolated cat heart. Previous in vitro studies had provided evidence to show that papillary muscles obtained from cats with chronic right ventricular failure had lost their ability to develop a positive inotropic response to glucagon. Since it is difficult to extrapolate from the isolated papillary muscle to the intact heart, studies were done to assess the effects of glucagon in the perfused isovolumically beating heart obtained from cats four months after surgical banding of the pulmonary artery for the experimental production of chronic right ventricular failure (CRVF). At the peak of the dose-response curve, glucagon increased right ventricular isovolumic pressure 25% (39.00 +/- 4.37 to 49.67 +/- 5.15 mm Hg; p less than 0.001) and right ventricular dP/dt 63% (522.2 +/- 93.9 to 852.6 +/- 159.9 mm Hg/sec; p less than 0.001) in 6 normal hearts. Similar dose related increases in right ventricular isovolumic pressure and dP/dt were obtained in 6 hearts taken from cats with chronic right ventricular failure. The respective increases in right ventricular isovolumic pressure and dP/dt were 43% (30.33 +/- 4.01 to 43.67 +/- 6.25 mm Hg; p less than 0.025) and 73% (317.50 +/- 30.29 to 550.83 +/- 89.04 mm Hg/sec; p less than 0.025). These results provide evidence that glucagon possesses the capacity to augment myocardial contractility in the heart with experimentally induced chronic right ventricular failure.


Comparative studies of a new 5HT-uptake inhibitor and some tricyclic thymoleptics. The new 5HT-uptake inhibitor, FG 4963, and some tricyclic thymoleptics antagonized p-chloroamphetamine (PCA)-induced hypermotility in rats. FG 4963 was active in about the same s.c. and p.o. doses as chlorimipramine. FG 4963, imipramine and chlorimipramine potentiated hypermotility induced in mice by the 5HT precursor 5HTP, FG 4963 being slighly more active than chlorimipramine. In contrast to the tricyclic thymoleptics FG 4963 did not potentiate the heart rate increasing effect of NA in pithed rats. The peripheral anticholinergic effect of FG 4963 and of desipramine was almost identical while the other imipramine derivatives were more active. All tricyclic thymoleptics were strong peripheral antihistaminics, but FG 4963 was almost devoid of this action. Acute tests for ECG changes in guinea pigs and toxicity in mice and rats showed that FG 4963 and chlorimipramine were less toxic than imipramine and amitriptyline. FG 4963 is presumably a selective 5HT-uptake inhibitor producing much less potentiation of peripheral sympathetic mechanisms than do the tricyclic antidepressants.


Evidence of central influences on blood glucose level: malathion hyperglycemia. To suppress the hyperglycemic effect of malathion in rats, a smaller amount of atropine was required when the drug was injected by intraventricular (i. vent.) than s.c. Pentobarbital, but not diazepam, blocked the hyperglycemic response. The results suggest that central accumulation of acetylcholine was the mediator of the response. Since hyperglycemia was not abolished by adrenalectomy and/or hypophysectomy, a hypothesis is presented to explain how central accumulation of acetylcholine might cause hyperglycemia.


Anatomical specificity as the critical determinant of the interrelationship between raphe lesions and morphine analgesia. Electrolytic lesions were produced in three separate parts of the raphe mesencephalic area: the nucleus raphe medianus, the nucleus raphe dorsalis and an area between these two nuclei. Seven days after the surgery, the animals were tested for morphine analgesia using the tail compression method and then sacrificed for the estimation of brain serotonin. It was found that the analgesic effect of morphine was significantly reduced in the rats lesioned in the nucleus raphe medianus but not in the animals lesioned either in the nucleus raphe dorsalis or in the 'intermediate raphe area'. Since a decrease of forebrain serotonin was observed in each experimental group, the reduction of morphine analgesia does not involve a simple direct correlation with a decrease of serotonin in the forebrain. The results are discussed in view of the possibility that the reduction of morphine analgesia after lesions of the nucleus raphe medianus is due to a disruption of a specific brain serotonergic system.


Apomorphine: facilitation of sexual behaviour in female rats. Sexual receptivity of spayed female rats was studied by recording lordosis responses to a mounting male. Administration of apomorphine (0.25 mg/kg) to oestrogen+progesterone-primed females produced an increase in lordosis behaviour 2--48 hr after injection. An increase was also found when apomorphine was substituted for progesterone. These results suggest a role for dopamine in controlling sexual receptivity.


Blockade of reserpine emesis in pigeons by metoclopramide. Reserpine (0.5 mg/kg i.m.) produced emesis in pigeons with 60% of the animals responding. Metoclopramide HCl at 10, 20 and 40 mg/kg p.o. administered 30 min before or after reserpine injection was effective in blocking reserpine emesis. Metoclopramide was unable to antagonize reserpine-induced sedation and hypotension in rats, thus inviting discussion of its possible mechanism in blocking reserpine emesis.


Tremor production by intracaudate injections of morphine. Pronounced resting tremor was produced in unanesthetized cats by injecting morphine (25-110 mug) into the caudate nucleus. The effects of morphine were antagonized by intracaudate (i.c.) injections of nalorphine (81-263 mug). Tremor activity was also inhibited by i.c. injections of dopamine (61-145 mug), Ca2+ (24-40 mug), scopolamine (88-121 mug) and hemicholinium-3 (HC-3; 73-129 mug) while serotonin (125 mug) was ineffective. Tremor inhibition by HC-3 was reversed by i.c. doses of acetylcholine (15-30 mug) which were subthreshold for tremor production in the absence of morphine. Morphine (55-110 mug) further increased the intensity of ongoing tremor activity induced by physostigmine (111 mug i.c.) I.c. injections of nalorphine antagonized the motor effects of morphine without affecting physostigmime tremor. Tremor production by morphine is attributed to a reduction in dopamine function which allows cholinergic activity in the caudate nucleus to predominate.


Tricyclic antidepressants: effects on the firing rate of brain noradrenergic neurons. The spontaneous activity of the norepinephrine-containing cells of the locus coeruleus was recorded in chloral hydrate-anesthetized rats. The effect of seven tricyclic antidepressants on the firing rate of single cells in the locus coeruleus was studied. All the drugs tested, except iprindole markedly decreased the rate of firing of the noradrenergic cells. Antidepressants having a secondary amine in the side chain, desipramine, nortriptyline and chlordesipramine, were more potent than their respective tertiary amine analogues, imipramine, amitriptyline and chlorimipramine. Alteration of the rate of drug metabolism by pretreatment with SKF-525A or phenobarbital did not change  the doses of tertiary antidepressnats required to decrease norepinephrine cell firing. Depletion of the norepinephrine stores by pretreatment with alpha-methyl-p-tyrosine and reserpine markedly increased the dose of desipramine required to depress the norepinephrine cells. The results are in good agreement with previous studies showing that secondary amine antidepressants are more potent than their tertiary amine homologues in blocking the uptake of norepinephrine into brain and peripheral tissues. Despite their lower potency it is concluded that tertiary antidepressants act on noradrenergic neurons in their unchanged form and not via secondary amine metabolites formed during the recording experiments since alterations in liver metabolism did not influence the response. The findings are consistent with the suggestion made from studies on transmitter turnover that antidepressants by inhibiting reuptake of norepinephrine cause a stimulation of postsynaptic receptors which decreases the activity of the presynaptic neurons by a feed-back mechanism. This view is further supported by the finding of an inverse relation between the norepinephrine content of the brain and the dose of desipramine required to decrease the firing rate of the noradrenergic neurons.


Influence of reserpine pretreatment on cardiovascular effects of angiotensin II in rabbits. Angiotensin II induced a dose-related increase in the amplitude of contraction of preparations of isolated atria and aortic strip of rabbit. These effects were significantly higher in organs obtained from reserpine pretreated animals (4 mg/kg i.p. 24 hr before experiments) than in controls. After reserpine, the effect of four doses of angiotensin II on the blood pressure of rabbits under pentobarbital anesthesia was also significantly higher than in untreated controls.


Apomorphine as a discriminative stimulus, and its antagonism by haloperidol. At a non-stereotypogenic dose (0.16 mg/kg, s.c.), apomorphine was shown to produce a discriminative stimulus in rats. The apomorphine-produced stimulus was found to be effectively antagonized by haloperidol. It is suggested that apomorphine's ability to stimulate central dopamine receptors is directly related to its cueing action.


Comparison of the inhibitory effects on the guinea-pig taenia coli of adenine nucleotides and adenosine in the presence and absence of dipyridamole. Dipyridamole potentiated the inhibitory responses to ATP and ADP and in particular responses to AMP and adenosine. The results are discussed in relation to the known actions of dipyridamole and suggest that adenine nucleotides are metabolized to adenosine during contact with the preparation; and that previous observations of the relative potencies of adenine nucleotides and nucleosides do not represent the actual receptor agonist potencies because of rapid uptake of adenosine into the tissues.


Influence of reserpine pretreatments on cardiovascular effects of angiotensin II in rabbits. Angiotensin II induced a dose-related increase in the amplitude of contraction of preparations of isolated atria and aortic strip of rabbit. These effects were significantly higher in organs obtained from reserpine pretreated animals (4 mg/kg i.p. 24 hr before experiments) than in controls. After reserpine, the effect of four doses of angiotensin II on the blood pressure of rabbits under pentobarbital anesthesia was also significantly higher than in untreated controls.


Comparison of the effects of pentobarbital and chloralose on the cardiac rhythms of the dog in chronic atrio-ventricular heart block. In the dog in chronic atrio-ventricular dissociation, pentobarbital at a dose of 25 mg/kg causes atrial tachycardia, but does not change the ventricular frequency. The vagolytic effect of pentobarbital explains this phenomenon. The pronounced vagal tonus seen in the Keith-Flack node, is not seen in the ventricle. Chloralose at a dose of 80 mg/kg i.v. does not alter atrial frequency but depresses ventricular frequency, in a significant way, for 40 min.


Cardiovascular effects of diazepam and chlordiazepoxide. In open-chest dog preparations i.v. administration of diazepam and chlordiazepoxide caused increases in coronary blood flow and cardiac output while reducing blood pressure, heart rate, and myocardial contractile force. The decrease in force was independent of the alteration in coronary flow. Arterial--coronary sinus oxygen difference narrowed and myocardial oxygen consumption decreased. These data show that both diazepam and chlordiazepoxide exert an oxygen conserving action along with an increase in myocardial oxygen delivery and suggest that these drugs may be of benefit to the patient with coronary insufficiency by a direct effect on the cardiovascular system as well as through CNS-mediated antianxiety effects.


The importance of extrapyramidal function for the induction and antagonism of harmine tremor. The brain lesion technique was used to investigate the role of the paleostriatum and the nigro-neostriatum in harmine-induced tremor and in its antagonism by dopaminergic agonists, apomorphine, 1-dopa, piribedil, d- and l-amphetamine. Bilateral lesions of the caudate--putamen or substantia nigra failed to modify the intensity of tremor or its antagonism by dopaminergic agonists. Bilateral lesions of the globus pallidus markedly reduced the intensity of tremor and the results indicated that such lesions were also able to reduce the effectiveness of dopaminergic agonists as tremor antagonists. The data suggest that the integrity of the paleostriatum is more important than that of the neostriatum for the mediation of harmine tremor and its antagonism by dopaminergic agonists. The results are discussed in relation to the proposed clinical relationship between paleostriatal dopamine dysfunction and tremor mechanisms.


Loss of body weight as a predictor of reserpine-induced amine depletion. Loss of body weight was proved to be a convenient and reliable way to predict the degree of reserpine-induced amine depletion after i.p. injections of reserpine (2.5, 5 and 10 mg/kg) and it thus helps reduce variability. Two populations of animals were sharply distinguished lying on either side of the 5% cutoff point within the first 24 hr. One group (approximately 20% of all animals) lost less than 5% of their body weight, showed absence of physiological and behavioural effects, a moderate 5-HT depletion and a moderate 5-HIAA increase. These animals were called 'partially reserpinized'. Another group always lost 5% or more of body weight, exhibited the reserpine syndrome as well as a larger extent of 5-HT depletion and 5-HIAA increase. This group comprised 'reserpinized' rats. There were significant correlations between the percent weight loss and the degree of 5-HT depletion and 5-HIAA increase in the group of 'partially reserpinized' animals. The two groups differed also in the time course of amine changes. Dose had no effect in the differentiation of the two groups but did affect the extent of amine depletion. When reserpine was given i.v. or s.c., all animals were 'reserpinized'. The phenomenon of partial reserpinization is discussed.


The restorative effect of erythropoietic stimulation upon the sublethally irradiated (SLI) hematopoietic stem cell and/or its progeny. Bone marrow cells from sublethally irradiated donor mice injected into supralethally (1000 R) irradiated recipients produced not only many fewer spleen colonies but fewer erythrocytic (E) colonies in relation to the number of granulocytic (G) colonies than did unirradiated donor marrow in similar recipients. This reversal of E:G ratio was partially or completely restored by a strong erythropoietic stimulus in the recipients, whether endogenous, (induced by anemia) or exogenous (injected purified EP). Such a stimulus produced a large increase in total spleen colony numbers in all experiments. An unexpected result was a major increase in granulocytic and megakaryocytic colonies in the spleen and marrow of the erythropoietically stimulated recipients. It was concluded that the sublethal irradiation to the donor either produced or revealed a subpopulation of potential colony forming cells whose cell cycle was slow or temporarily arrested. The added EP (and/or related substances) was proposed to have either overridden or induced repair or radiation defect, or stimulated a small and normally undetectable population requiring high levels of stimulant to induce cycling. The result was the expression of their progeny as colonies, whose differentiative type depended upon the kind of hematpoietic inducing microenvironment in which they had lodged.


Labyrinthine and vestibulospinal effects on spinal motoneurons in the pigeon. Experiments were performed on pigeons to investigate the effects of labyrinthine stimulation on motoneurons innervating neck and limb muscles. Intracellular recordings from the ventral horn of the spinal cord in the neck revealed that stimulation of the ipsilateral labyrinth with only a single shock evokes prominent EPSPs and/or IPSPs in the majority of neck motoneurons at thresholds less than 3--4 times the threshold for the labyrinth-evoked N1 potential recorded in the ipsilateral vestibular nuclei. In many neck motoneurons, the latencies of the PSPs were short enough so that no more than two synapses could be involved in their transmission. Recordings were also obtained from identified motoneurons which innervate muscles of the wing or the leg. Stimulation of the labyrinth, even with multiple stimuli, failed to elicit any observable PSPs in limb motoneurons. These data were consistent with other observations that in awake pigeons stimulation of the labyrinth with weak shocks evoked a pronounced turning of the head to the contralateral side, while even strong or multiple stimuli failed to produce any observable movement of wings or legs. Thus in the pigeon the association of the labyrinth with limb muscles is insignificant when compared to the association of the labyrinth with neck muscles. In addition, supraspinal descending fibers in the pigeon may make monosynaptic excitatory and inhibitory contact with spinal motoneurons.


Autoradiographic localization of 3H-gamma-aminobutyric acid in the medial hypothalamus. Tritium labelled gamma-aminobutyric acid (3H-GABA) was infused into the third ventricle of rats with normal or deafferented hypothalamus and the distribution of the label was studied by light and electron microscopic autoradiography. In control as well as deafferented hypothalamus a few neurones accumulated radioactivity, while the majority was unlabelled. Characteristic clusters and rows of silver grains were observed in the neuropil of several regions probably indicating labelled cell processes and terminal axons. Electron microscopy showed that at least some of the clusters were over axon terminals with synaptic vesicles. 3H-GABA accumulated also in the ependyma and glial elements. The results suggest that in the medial hypothalamus there is a preferential uptake of GABA in some neurones and nerve fibers; at least some of these are hypothalamic interneurones. This supports the hypothesis that some hypothalamic neurones and nerve endings may use GABA as a transmitter.


The dorsal column system: I. Existence of long ascending postsynaptic fibres in the cat's fasciculus gracilis. Microelectrode recordings were made from dorsal column fibres at the 12th thoracic segment in cats. Two kinds of activity were elicited by electric shocks applied on the sciatic nerve in the popliteal fossa: primary afferent activity and trans-synaptically evoked activity. The dorsal column postsynaptic (DCPS) fibres represented 9.3% of the total population of fibres studied in the fasciculus gracilis and 14.5% of those with receptors in the skin. They were found to lie between the primary fibres of cutaneous and those of deep origin. The fastest fibres of the alpha range contributed to their activation and it is likely that C fibres contributed as well. 87% of the DCPS fibres studied at Th 12 were antidromically activated from the first cervical segment, and their conduction velocities measured between cervical and thoracic levels ranged from 16 to 71 m/sec.


The dorsal column system: II. Functional properties and bulbar relay of the postsynaptic fibres of the cat's fasciculus gracilis. Microelectrode recordings were made from dorsal column postsynaptic (DCPS) fibres, in the fasciculus gracilis of the cat, at thoracic level Th 12, and from single cells in the nucleus gracilis. The sensitivity of the fibres (Th 12) and cells (bulbar level) to both gentle and noxious stimuli was studied and a classification of the units was made on the basis of their responses to these natural stimuli. The DCPS fibres have been classified into three groups: i) 16.3% of them were activated only by light mechanical stimuli, ii) a few responded to nothing but noxious mechanical stimuli, iii) 77% were characterized by their sensitivity to both gentle and noxious stimuli and constituted the polymodal group. The study of the nucleus gracilis cells revealed that 31.2% of the total number of cells investigated in the nucleus were also characterized by a high degree of modality convergence (polymodal cells). Besides their convergence, the polymodal DCPS fibres and the polymodal cells of the nucleus gracilis were functionally in contrast to the properties of the specific fibres and cells of the dorsal column system in other respects: i) they responded with a slowly adapting response to constant mechanical stimulation of the skin, ii) they were sensitive to noxious mechanical and thermal stimuli. It is argued that the polymodal cells could be the bulbar relay of the impulses conveyed through the DCPS fibres of the fasciculus gracilis. The possible role of the dorsal column system in nociception is discussed in the light of the results.


Response of medial preoptic neurons to electrical stimulation of the mediobasal hypothalamus, amygdala and mesencephalon in normal, serotonin or catecholamine deprived female rats. Single cell activity from preoptic neurons was extracellularly recorded in normal female rats and the effects of electrical stimulation of the mediocortical amygdala (AMY), the N. med. raphes (MES) and the mediobasal hypothalamus (MBH) was tested. 1. One type of preoptic neurons (55%) reacted with primary excitation to a single electrical stimulus of AMY, MES or MBH, which was usually followed by a period of postexcitatory inhibition. Another type of neurons (29%) were first inhibited after stimulation of AMY, MES or MBH and then showed a period of postinhibitory excitation. The discharge rate of a third type was not affected by electrical stimulation of these structures (16%). If a neuron was affected by a stimulus in a given area it predictably reacted in the same fashion to stimulation of the other areas. 2. The postexcitatory inhibition of the majority of the primarily excited neurons was strong enough to prevent the stimulus response to a second stimulus. Occasional recordings from two neighboured cells simultaneously indicate that they can be inhibitory to each other. 3. By pulse train stimulation of the AMY or MES with varying frequencies it could be demonstrated that low frequencies (10 Hz) had a facilitatory action whereas higher frequencies (100 Hz) were inhibitory to preoptic discharge rates. 4. No obvious alteration of neuronal properties we found neither in rats treated intraventricularly with 5,6-dihydroxytryptamine, which strongly reduces central nervous system serotonin content, nor in 6-hydroxydopamine treated rats, which had low central nervous system catecholamine levels.


Quantitative studies on the supraoptic nucleus in the rat. I. Synaptic organization. Four types of synapses: axo-axonic, axo-dendritic, axon-spine, and axo-somatic were distinguished in the supraoptic nucleus in the rat. The density of synaptic terminals (boutons) in this nucleus is 35.17-10(6)/mm3 hence there are over 5 million boutons on each side, and on the average 596 per neuron. Only about one third of the axon terminals in this nucleus originate outside the nucleus and its immediate neighbourhood. Two thirds appear to be of intranuclear or otherwise of local origin. This could be explained by assuming either numerous intranuclear axon collaterals or interneurons having richly arborizing axons, or possibly both.


Quantitative studies on the supraoptic nucleus in the rat. II. Afferent fiber connections. A quantitative electron microscopic study of synaptic terminal degeneration was performed in the supraoptic nucleus (NSO) after a variety of major transections or ablations, destroying or interrupting in different combinations the afferent pathways known from earlier and own light microscopic degeneration studies. Solutions of a set of equations, expressing the percentage degenerations in synaptic profiles after different combinations in which the several pathways are interrupted by the various interferences, enabled the authors to give the following percentage numbers for afferent synapses from different sources. 32.7% of supraoptic afferents originate from the brain stem probably representing the monoaminergic innervation of this nucleus. The medial basal hypothalamus (21.0%), amygdala (13.5%), septum (13.5%), hippocampus (8.5%) and olfactory tubercle and further rostral cortical region (17.0%) are the other main sites of origin of supraoptic nucleus afferents. There are no supraoptic afferents from the optic nerve, superior cervical ganglion or fimbria hippocampi.


Dorsal column nuclei afferents in the lateral funiculus of the cat: distribution pattern and absence of sprouting after chronic deafferentation. In adult cats the successive degeneration technique has been used to demonstrate the existence and distribution pattern of lateral funicular fibers to the dorsal column nuclei (DCN) originating from the brachial and thoracic cord. In a first operation, interruption of the dorsal columns at appropriate cervical levels and of the lateral funiculus at low thoractic levels was performed. Thirteen months later, a lesion was made in the lateral funiculus at upper brachial or uppermost thoracic levels. Fiber degeneration in the DCN consequent to this second operation is not contaminated by damage to dorsal roots or by interruption of lateral funicular afferents from lumbo-sacro-coccygeal segments. All animals were sacrificed 7 days after the second operation. Serial sections through the medulla oblongata, impregnated with the Fink-Heimer technique, show that fibers ascending from brachial levels in the dorsal part of the lateral funiculus reach the cuneate nucleus either by a dorsomedial route through the tegmentum or by cuneate nucleus either by a dorsomedial route through the tegmentum or by ascending in the restiform body. Degenerated fibers distribute selectively to the rostral part, and to a lesser extent to the base of the cuneate nucleus. Only very few fibers ascending from thoracic levels in the lateral funiculus distribute to the DCN. In another group of animals, not previously deafferented, a lesion of the lateral funiculus was made at upper brachial levels. This group served as a control to assess whether sprouting had occurred in the chronic preparations as a consequence of the long-term deafferentation. Comparison of the results in the cuneate nucleus of the two groups of animals shows no difference in the pattern of distribution or in the amount of degenerated fibers in this nucleus. These observations are discussed in relation to the question of collateral sprouting in the adult mammalian central nervous system.


Eye and head movements evoked by electrical stimulation of monkey superior colliculus. In unrestrained animals of many species, electrical stimulation at sites in the superior colliculus evokes motions of the head and eyes. Collicular stimulation in monkeys whose heads are rigidly fixed is known to elicit a saccade whose characteristics depend on the site stimulated and are largely independent of electrical stimulation parameters and initial eye position. This study examined what role the colliculus plays in the coding of head movements. A secondary aim was to demonstrate the effects of such electrical stimulation parameters as pulse frequency and intensity. Rhesus monkeys were free to move their heads in the horizontal plane; head and eye movements were monitored. As in previous studies, eye movements evoked by collicular stimulation were of short latency, repeatable, had a definite electrical threshold, and did not depend on the initial position of the eye in the orbit. By contrast, evoked head movements were extremely variable in size and latency, had no definite electrical threshold, and did depend on initial eye position. Thus when the eyes approached positions of extreme deviation, a head movement in the same direction became more likely. These results suggest that the superior colliculus does not directly code head movements in the monkey.


Recurrent inhibition of individual Ia inhibitory interneurones and disinhibition of their target alpha-motoneurones during muscle stretches. The effects of ramp stretches applied to triceps surae muscle on the discharge patterns of single Ia inhibitory interneurones, monosynaptically invaded from various nerves, were studied in either decerebrate or anesthetized cats. Interneurones which received direct excitatory Ia input from the stretched muscle exhibited augmented activity both during the dynamic and static phase of stretch, which was, however, interrupted by a transient inhibitory influence during the dynamic phase of stretch. The influences on Ia inhibitory interneurones, monosynaptically invaded from hamstring or tibial nerve, were exclusively inhibitory. These stretch-induced inhibitions were better demonstrable in decerebrate than in anesthetized preparations. The timing of the discharge patterns of additionally recorded Renshaw cells during stretch, and the disappearance or reduction of the above described inhibitory effects after administration of DHE, strongly support the idea that these inhibitory actions are caused by Renshaw inhibition. In Ia inhibitory interneurones, monosynaptically activated from the antagonistic peroneal nerve, stretch induced also pronounced inhibitory effects, which were most probably caused by mutual inhibition between Ia inhibitory interneurones. The suppression of agonistic Ia inhibitory interneurone activity below the tonic resting activity corresponded to an enhancement of the monosynaptic reflex amplitude of the antagonistic motoneurone pool. The findings suggest that normal orthodromic activation of Renshaw cells, and consequently the recurrent inhibition of the Ia inhibitory interneurones, is predominantly linked with rapid phasic, rather than slow tonic, motoneuronal firing. The functional role of this mechanism for the performance of rapidly alternating movements and the damping of ballistic agonist contractions is discussed.


Dissociated cell cultures from fetal mouse hypothalamus. Patterns of organization and ultrastructural features. Dissociated fetal hypothalamic cells mainly taken from 14 day-old mouse fetuses were grown in vitro for increasing time (9 to 60 days). Soon after inoculation the cells partly reaggregated and attached. The small reaggregates were then interconnected by fibers bundles. After the first week the cultures were composed of a continuous basal monolayer of flat and transparent cells, over which various types of refractile cells were lying in discontinuous areas. The ultrastructural study enabled us to identify these cell types, to describe their spatial relationships, and to follow their evolution with time in culture. The basal cell formed several superimposed layers. With increasing age, they displayed typical features of astrocytes and of ependymal cells. The latter exhibited rhythmic ciliary movements in culture. The overlying cells corresponded to three types which were associated in small clumps: primitive neuro-epithelial cells, maturing as well as mature neurons and typical neurosecretory cells. The latter cells were found as early as 9 days of culture of 14 day-old fetal hypothalamic cells and retained their typical features up to two months. Neuronal processes formed very dense networks at the surface of the cultures and terminated within the basal layers. Axon and dendrites were precociously found and were still present after two months. Within axon terminals dense-core vesicles appeared at the same time as neurosecretory cells. Synaptic vesicles and synaptic junctions were found later on.


Electrical constants of neurons of the red nucleus. Membrane electrical constants have been studied in neurons of the red nucleus (RN) of the cat which were identified antidromically from the spinal cord. For each cell, the input resistance was determined from the membrane potential changes to current steps and was found to be 2.5+/-0.9 Momega in twenty five RN cells studied. In addition, linear summation of the membrane responses induced by two current pulses was demonstrated. 2. From the membrane voltage transients to current steps, the first membrane time constant, tau0, and second time constant, tau1, were determined as 5.6+/-1.0 msec and 0.6+/-0.2 msec, respectively. The ratio of the amplitudes of two exponential functions, E1/E0, was 0.18+/-0.05. A linear relation was found between the ratio of these amplitudes of exponential functions and that of the two time constants. 3. The cable parameter (electrotonic length, L) of the combined soma and dendrites of the RN neutrons was estimated as 1.1 from membrane transient data using the relation developed by Rall (1969). 4. By using this parameter, an attempt was made to estimate the location of the two excitatory inputs on the soma-dendritic membrane of RN cells.


Central inhibitory interactions in human vision. Contrast threshold and perceived orientation of a line segment were measured when another line segment was simultaneously presented either to the same or the other eye; the angle between the two line segments was varied. The presence of the masking line elevated the contrast threshold under both conditions and the threshold increased similarly both in monoptic and dichoptic masking when the masking angle was made smaller. The presence of the masking line affected also the perceived orientation of the test line. The effect was similar both in monoptic and dichoptic masking; the largest change of perceived orientation occurred at about 15 degrees masking angle and the change was smaller at other angles. The effect disappeared within a short distance when the masking line was removed farther from the test line. The similarity of the monoptic and dichoptic threshold elevations demonstrates that there are lateral inhibitory interactions between central neural units in the human visual system. It is likely that the interacting units mediate the perception of contour orientation, for the threshold elevation functions were consistent with concurrent changes of perceived orientation. The results are evidence for the hypothesis that inhibition between orientation detectors is a factor in the perceptual expansion of acute angles.


Axonal conduction latencies of cat retinal ganglion cells in central and peripheral retina. The axonal conduction latency of brisk-sustained and brisk-transient cat retinal ganglion cells increases with proximity to the area centralis. The trend is gradual with considerable local scatter of latencies, and is similar for cells with crossed and uncrossed axons.


Electrophysiological studies of the projections from the parietal association area to the cerebellar cortex. 1. Responses evoked in the cerebellar cortex by stimulation of the parietal association cortex (rostral portions of the middle suprasylvian gyrus) were recorded and analysed in cats, and were compared with those by stimulation of the motor cortex (anterior sigmoid gyrus). 2. The parietal stimulation elicited early mossy fibre and late climbing fibre responses in the cerebellar cortex. The mossy fibre responses appeared at a latency of 2.0--2.5 msec and predominantly in the lateral (hemispherical) part of the contralateral cerebellum (mainly crus I, crus II and paramedian lobules). Cutting of the inferior cerebellar peduncle produced little or no influence upon the mossy fibre responses, which suggests that the mossy fibre responses are mediated chiefly by the pontine nuclei. 3. The climbing fibre responses were recorded at a latency of 17--19 msec and markedly in the contralateral intermediate and medial parts of IV--VI lobules. The responses were easily sppressed by anaesthesia and depended on the conditions of experimental animals. The unstable appearance of the responses and their longer latencies than those of the climbing fibre responses due to stimulation of the motor cortex imply indirect pathways from the parietal association cortex to the inferior olive. 4. The predominant projection of the parietal-induced mossy fibre responses to the lateral part of the cerebellum was compared with the mossy fibre projection from the motor cortex and was discussed as an important component in the cerebrocerebellar loops.


[Pyrazino(1,2-a)- and 1,4-diazepino(1,2-a)- indoles. III. Synthesis of pyrrolo(2',1':3,4)pyrazino(1,2-a)indoles and pyrrolo(2',1':3,4)-1,4-diazepino(1,2-a) indoles; isoindolo(2',1':3,4)pyrazino(1,2-a)-indoles and isoindolo(2',1':3,4)-1,4-diazepino(1,2-a)indoles; pyrazino(3,2,1-jk)- carbazoles and 1,4-diazepino(3,2,1-jk) carbazoles]. Pyrrolo(2',1':3,4)pyrazino (or 1,4-diazepino) (1,2-a)indoles and isoindolo(2',1':3,4)pyrazino (or 1,4-diazepino) (1,2-a)indoles were synthesized by reaction of 1-(2-aminoethyl)indole or 1-(3-aminopropyl)indole with gamma-keto and o-acylbenzoic acids. In addition, a novel synthetic route for the preparation of pyrazino(3,2,1-jk)- and 1,4-diazepino(3,2,1-jk)carbazoles has been realized.


[Cyclic hydrazides. II. (1) Synthesis of 3-aminopyrazolo[1,2-b]hthalazine-1(1H),5(10 H)-diones]. The synthesis is described of a number of 3-aminopyrazolo[1,2-b]phthalazine-1(1H),5(10H)-diones (I) differently substituted on the amino group and in position 2, through condensation of alpha-chloro-beta-chlorocarbonylenamines (IV) with the 3,4-dihydrophthalazin-1(2H)-one (III). Alternatively cyanoacetylchlorides are condensed with (III) giving directly the 3-aminopyrazolo[1,2-b]phthalazine-1(1H),5(10H)-diones with the primary amino function (VII).


[Derivatives of 1,5-bis-(3,4-methylendioxyphenyl)-n.pentane myolytic activity]. A series of amines was prepared from 1,5-bis-(3,4-methylendioxyphenyl)-n.pentan-3-one by reduction of the oxime, by the Leuckard-Wallach reaction or via the Grignard reaction with 3-dialkylamino propylchlorides. These substances proved to have myolytic activity.


[Hexahydroimidazo[1,5-a]pyrazine. I. Synthesis of 7-methyl-1,5,6,7,8a-hexahydroimidazo[1,5-a]pyrazin-3(2H)-one and derivatives]. The synthesis and pharmacological activity of a series of 2-aryl or alkyl substituted 7-methyl-1,5,6,7,8,8a-hexahydroimidaso[1,5-a]pyrazin-3(2H)-ones, are reported. The 2-aryl derivatives (VI) have been prepared by reaction of 3-(arylaminomethyl)-1-methylpiperazines (IV) have been prepared by reaction of 3-(arylaminomethyl)-1-methylpiperazines (IV) with N,N'-carbonyldiimidazole. Reaction of various anilines with 3-carbomethoxy-1-methylpiperazine (II) and subsequent reduction of the amides (III) afforded the bases (IV). The synthesis of the unsubstituted compound (XII) has been accomplished either by cyclization with sodium methoxide of methyl (1-me-thylpiperazin-3-yl)methylcarbamate (XI) or by reaction of 3-aminomethyl-1-methylpiperazine (XV) with N,N'-carbonyldiimidazole. Reduction of 1-benzyl-2-cyano-4-methylpiperazine (VII) followed by reaction with methyl chloroformate and debenzylation afforded the urethane (XI). The 2-alkyl and 2-alkenyl derivatives (XIII) have been prepared by alkylation of the sodium salt of (XII) in DMF. The compounds of these series have been tested for antiinflammatory, coronary dilator and C.N.S. depressant activities.


[Analysis of the hypolipemic, hypocholesteremic and platelet anti-aggregating effects of tibric acid]. Tibric acid shows interesting hypocholesteremic and hypolipemic activity greater than that of clofibrate. In addition high concentrations of tibric acid affect platelet aggregation. Tibric acid does not show choleretic activity and does not affect liver function.


[Derivatives of imidazo[1,2-c]quinazoline with inhibiting effect on platelet aggregation]. Several 5-monosubstituted and 5,5-disubstituted 5,6-dihydroimidazo[1,2-c]quinazolines were synthesized in high yields by the condensation of 2-(o-aminophenyl)imidazole with the required aliphatic or aromatic aldehydes or ketones. Some of these compounds caused inhibition of ADP-induced platelet aggregation in vitro at concentration of 10(-5) to 10(-4) M. The structure-activity relationships of these inhibitors are discussed.


[Steroidal heterocyclic compounds: [16,15-c]-pyrazole derivatives of estradiol and of estrone]. Condensation between ethyl formate and estradiol 16-keto derivatives, in which the 17beta-hydroxyl groups had been protected as tetrahydropyranyl ethers, produced the corresponding 15-hydroxymethylene compounds. These, by cyclization with hydrazine, gave the [16,15-c]-pyrazole derivatives. Further reactions, such as hydrolysis, etherification and chromic oxidation, afforded many differently substituted [16,15-c]-pyrazoles of the estradiol and estrone series.


[Research on substances with antiviral activity. III. Structure of the monosubstituted derivatives of 1 H-benzo-[e] indene-1,3(2H)-dione]. The N.M.R. spectra of 1H-benz[e]indene-1,3(2H)-dione (I), 1H-benz[e]indene-2,3-dihydro-1-one (II), 7-methyl-3H-benz[e]indene-1,2-dihydro-3-one (III) and of their oximes, phenylhydrazones and 4-methylthiosemicarbazones show the preferential substitution at 3 position for monoderivatives of (I). This is very interesting in correlation with the structure-activity relationship of derivatives of (I) and (II), some of which have shown in vitro antiviral action in previous research.


[Research on substances with antiblastic activity. LVIII. Synthesis and antineoplastic activity of derivatives of pyrrolo-(1,2-a)quinoxaline]. Some derivatives of pyrrolo[1,2-a]quinoxaline and 7,8-dimethoxypyrrolo[1,2-a]quinoxaline, structurally related to two antitumoral antibiotics, mitomycin C and anthramycin, have been prepared. The synthesis of the two nuclei was carried out starting from the required nitroaniline and 2,5-diethoxytetrahydrofuran; formylation of the pyrrole derivative and successive reduction gave the heterocyclic compound directly. Antineoplastic activity was evaluated on mice treated intraperitoneally with leukemia L 1210.


Second-order schedules of drug injection. Key-press responding of squirrel monkeys produced intravenous injections of cocaine under two simple types of schedule. Under a fixed ratio schedule, every 30th response produced an injection; steady responding at high rates of over one per second were maintained during each fixed-ratio component. Under a fixed-interval schedule, the first response occurring after a fixed time of 5 min produced an injection; there was a pause at the start of each interval and then progressively increasing responding until cocaine was injected at the end of the interval. Both squirrel monkeys and rhesus monkeys also were studied under second-order schedules of drug injection. Under one type of second-order schedule, studies only in squirrel monkeys, completion of each fixed-interval component produced only a 2 sec light; completion of the 10th fixed-interval component produced the brief light and an intravenous injection of cocaine. Under a second type of second-order schedule, each fixed-ratio component completed during a fixed time interval (5 or 60 min) produced only a 2-sec light; the first fixed-ratio component completed after the interval of time elapsed produced the brief light and an intravenous (squirrel monkeys) or intramuscular (rhesus monkeys) injection of cocaine. Under both types of second-order schedules, repeated sequences of responding were maintained during each session and characteristic fixed-interval or fixed-ratio patterns of responding were controlled by the brief visual stimuli.


Experimental models for the modification of human drug  self-administration: Methodological developments in the study of ethanol self-administration by alcoholics. Experimental studies of human ethanol self-administration are reviewed, and a description is provided of the procedural evolution that has oc-urred in the experimental study of the determinants of human ethanol self-administration. Human experimental models of alcoholism have been established within residential laboratories which permit chronic availability of ethanol to volunteer alcoholic subjects. Experimentation within such environments has progressed from observational and descriptive studies of experimental intoxication to studies that manipulate experimental variables so as to modify (reduce) ethanol self-administration by alcoholic subjects. To observe systematic effects of manipulated variables it has been necessary to develop sensitive baselines of ethanol self-administration. When ethanol intake has been relatively unrestricted, wide spontaneous fluctuations have made difficult the evaulation of manipulated variables. When a variety of restrictions on ethanol availability have been imposed, sensitive self-administration baselines have been established which have permitted the direct experimental assessment of some of the determinants of ethanol self-administration. Six methodological principles are suggested for enhancing the information yield of future research on the determinants of ethanol self-administration. The same general methodology is suggested for research with other varieties of drug self-administration.


Neurochemical changes associated with schedule-controlled behavior. Studies of monoamine metabolites within the cerebrospinal fluid compartment have indicated that this approach may be useful in examining central metabolic changes in vivo. By combining the technologies of radioisotope chemistry, operant behavior control and modification, and brain perfusion with push-pull cannulas, we have been able to examine minute to minute changes in the disposition of radiolabeled monoamine transmitter candidates and their metabolites. These substances appear to co-vary with changes in complex behavior maintained by operant schedules of reinforcement and affected by changes in schedules or administration of psychotropic drugs. In agreement with other perfusion studies, we have observed changes in fractional distribution of radiolabeled urea, a so-called extracellular marker, along with shifts in monoamines; but the former appear more transient. These observations nevertheless support the concept of dynamic changes within the extracellular environment of the CNS that may be part of a hormone-like communicating system with functional significance. Furthermore, the presence of peaks and/or troughs, in perfusates of [14C]urea of similar substances should not be taken as a priori evidence for nonspecificity of the technic, since selective release or inhibition of release of monoamines can be shown with appropriate drugs that are thought to act through these aminergic systems. Destruction of catecholamine nerve terminals with 6-hydroxydopamine likewise attenuates the signal-locked release of radiolabeled norepinephrine by a conditioned stimulus after conditioning occurs. No such release is seen on presentation of the to-be-conditioned neutral stimulus in control or 6-hydroxydopamine treated rats. These initial studies indicate the availability of a powerful tool for the study of drug-neurochemical-behavioral interactions using subjects as their own controls for extended periods of time so that phenomena of plasticity, tolerance and dependence may likewise be examined.


Behavioral effects of mercury and methylmercury. Intoxication by elemental mercury or by methylmercury is revealed primarily by changes in behavior and by neurological signs. Disorders of movement and posture have been most widely reported, both in animal experiments and in cases of human exposure. Specific sensory symptoms are also prominent in human methylmercury poisoning. Recent data indicate similar symptoms in monkeys during long-term exposure to methylmercury. Similar sensory  impairment has not been described in experiments with subprimates. Variations in the profile of behavioral and neurological effects are discussed in terms of differences in species and differences between acute and long-term exposure. The latter condition poses the most difficult questions for human health, yet has been less frequently studied. Procedures are suggested that may help to revolve these problems. In particular, tests of learned behavior hold great promise toward identifying specific symptoms and toward understanding how mercury compounds affect behavior.


Schedule-controlled behaviors as determinants of drug response. Schedule-controlled performances began to be used in assessing the behavioral effects of drugs because of practical advantages over other techniques for studying behavior. Schedule-controlled behavior is, however, of fundamental importance in behavioral pharmacology. It has been found repeatedly that the effects of many drugs depend critically upon the patterns of responding engendered by different schedule contingencies. These dependencies of the effects of drugs on schedule-controlled behavior occur because ongoing behavior is itself an important determinant of drug action.


Discrete trial analysis of drug action.

Discrete trial analysis of drug action. Discrete trial procedures permit exact control or description of the time of occurrence of stimuli, the probability of response occurrence, and the patterning of responses. They also make possible the experimental manipulation of the composition of the stimuli controlling behavior. The use of discrete trial procedures is illustrated here in an examination of the effects of scopolamine, a representative cholinergic blocker, on several aspects of behavior: Memory. Response alternation experiments, in which the spacing of discrete trials varies within the experimental session, show that, whereas accuracy of responding is consistently poorer under drug, the decline of accuracy with time since last trial is similar for drugged and nondrugged animals. Thus the drug does not affect memory "storage". Inhibition. Experiments in which discrete trials are presented in pairs, such that the correct response on Trial 2 of the pair is contingent upon Trial 1 events, show how the "disinhibiting" effect of scopolamine (as indicated by enhanced responding on "no go" trials) is augmented by increasing the time gap between Trial 1 and Trial 2, or by minimizing controlling stimuli on Trial 1. Discrimination. A variety of experiments suggest that scopolamine decreases the "detectability" of stimuli. Detectability effects, along with disinhibition observed under certain specific conditions, constitute the prinicipal behavioral actions of scopolamine observed with discrete trial procedures.


Gonadotropin levels and secretory patterns in patients with typical and atypical polycystic ovarian disease. Gonadotropin levels and secretory patterns were studied in 28 oligomenorrheic patients with various types of polycystic ovary disease (PCO). On the basis of ovarian morphology and histology, the patients PCOuld be separated into two distinct categories arbitarily designated "typical" (type I) and "atypical" (type II) PCO. Although no differences were noted in symptomatology or 17-ketosteroid, testosterone, or follicle-stimulating hormone levels, the 12 type I patients demonstrated widely fluctuating, but markedly elevated, luteinizing hormone (LH) levels, while the 16 type II patients demonstrated lower and less fluctuating LH levels which were comparable to those found during the normal follicular phase. It is likely that type I PCO is a distinct entity similar to that described by Stein and Leventhal, while type II co represents a heterogenous spectrum of disorders, many of which remain obscure.


Comparison of three intrauterine contraceptive devices: the Antigon-F, the Ypsilon-Y, and the Copper-T 200. A comparative study of three intrauterine contraceptive devices, the Antigon-F, the Copper-T, and the Ypsilon-Y, was carried out at The New York Hospital Cornell Medical Center during an 18-month period from March 1, 1972, through August 31, 1973. At the cutoff date of February 28, 1974, all patients had been followed for at least 6 months. During the period of study, 884 Antigon-F devices, representing 14,436 woman-months of use, were inserted. The over-all pregnancy rate was 0.88, with an explusion rate of 5.1 and a medical removal rate of 9.1/100 woman-years. This device was best tolerated by multiparous women. There was a total of 910 insertions of the Ypilson-Y, representing 14,348 woman-months of use. The over-all pregnancy rate was 1.8, with an explusion rate of 1.9 and a medical removal rate of 3.7/100 woman-years. The pregnancy rate in nulliparous women was 0.78, with a continuation rate of 88.9. The pregnancy rate in multiparous woman was higher, i.e., 2.4. Nine hundred thirty-nine Copper-T 200 devices, representing 15,558 woman-months, were inserted. The over-all pregnancy rate was 0.85, with an expulsion rate of 1.8, a medical removal rate of 4.5, and a continuation rate of 88.4/100 woman-years. The pregnancy rate was 0.82 for nulliparous women and slightly higher, i.e., 1.04, for multiparous women. Thus, all three devices had considerable merit. The Antigon-F and the Copper-T seemed the best in multiparous women; the Ypsilon-Y in the size used was best in nulliparous women. The only perforations occurred with the Copper-T.


In vitro fertilization of rabbit eggs in oviduct secretions from different days before and after ovulation. In vitro fertilization of rabbit eggs in oviduct secretions has been studied through different days of the psudopregnant cycle. The appearance of the pronuclei and cleavage into two cells, which were the criteria of fertilization, occurred significantly less frequently when in vitro fertilization was attempted in the oviduct secretions obtained during the estrous (preovulatory) period of the cycle than it was in the secretions obtained during pseudopregnancy or in Brackett's medium plus 20% heated rabbit blood serum. Whether fertilization was attempted in ampulla secretions or in whole oviduct secretions had no effect on the success rate. The effects of oviduct secretions from the estrous period of the cycle is probably due mainly to an effect on the egg, involving the very first development processes during and after fertilization.


Interruption of pregnancy by prostaglandin 15-methyl F2alpha. The current study was formulated to investigate the abortifacient activity of prostaglandin 15-methyl F2alpha (15-methyl PGF2alpha) administered intramuscularly to 80 healthy women with gestations between 8 and 22 weeks. Goals were the establishment of an effective dosage schedule and assessment of the incidence and severity of side effects. All 80 gravidas were aborted, with a mean time to abortion of 15.70 hours (SD, 6.52). Gastrointestinal side effects occurred in 89% of the patients; temperature elevations  greater than or equal to 100.6 degrees F were noted in 14 cases. No other significant complications were encountered. Transabdominal intra-amniotic pressure monitoring indicated the need to administer the drug at 2-hour intervals. The 15-methyl PGF2alpha patients were matched for parity and gestational length with 80 gravidas aborted with PGE2 20-mg vaginal suppositories. The difference in interval to abortion in the two groups was not statistically significant. While gastrointestinal side effects were more common with 15-methyl PGF2alpha, the frequency of drug-induced temperature elevations was reduced.


Human histocompatibility (HL-A) antigens in semen and their role in reproduction. Seminal plasma from 7 of 10 normal persons was found to inhibit anti-HL-A2, 3, 7, 8, and 12 antisera specifically. Inhibition by seminal plasma appeared quantitatively less than that by whole serum and even less than that by platelets from the same individuals. No decrease in the number of offspring was found due to increased numbers of HL-A incompatibilities of the male spouse in 67 normal couples. No clear decrease in the number of pregnancies was observed in normal women with lymphocytotoxic antibodies. A similar study with infertile persons may yet reveal a role for HL-A antigens in human reproduction.


Acrosin activity of cryo-preserved human spermatozoa. The acrosin activity of human spermatozoa was determined in fresh, glycerolated, and cryo-preserved semen specimens. In 119 semen specimens, mean increases in acrosin activity of 62% in glycerolated and 56% in cryo-preserved samples were found. Thus, no statistically significant differences in mean acrosin activity were found between glycerolated and cryo-preserved spermatozoa. However, 23.5% of the cryo-preserved samples showed a significant decrease in extractable acrosin activity when compared with untreated controls. In oligospermic specimens (less than 40 million spermatozoa/ml), a statistically significant decrease in acrosin activity due to cryo-injury was detectable. Differentiation of specimens responding to glycerol pretreatment with an increase in extractable acrosin activity from those responding with a decrease showed a different freezing behavior, indicating two ejaculate types with respect to acrosin extraction. Samples frozen without glycerol protection showed the same amounts of extractable acrosin activity as did glycerol-protected specimens. The stability of acrosin in acidic acrosomal extracts during liquid nitrogen freeze treatment was confirmed.


Psoriasis: a review. Part I.

Psoriasis: a review. Part I. The recent literature relating to pathogenesis, clinical features and management of psoriasis has been reviewed. The features of the accelerated epidermopoiesis are described and its relationship to a possible primary fault in keratinisation or dermal influences discussed. The present position on the genetics and epidemiology of psoriasis is summarised. Recent observations on the relationship of histocompatibility system antigens to ordinary and pustular psoriasis could lead to important advances. Clinical discussion is confined to psoriasis in childhood, the hair, nails and mucosal psoriasis; pustular psoriasis and psoriasis arthropathica.


Solitary papular lesions on the penis in insufficiently treated early syphilis. A report of two cases. Two cases of insufficiently treated early syphilis with solitary papular lesions on the penis are described. There seems to be a close resemblance between these cases and descriptions from the Salvarsan era concerning the occurrence of solitary papules located on or near the place of the healed primary syphilitic lesion, after insufficient treatment with Salvarsan.  Problems in diagnosing these cases are discussed.


Familial syringoma.

Familial syringoma. Syringoma occuring in two siblings in a South Indian family is reported. A familial occurrence of such an uncommon tumour is of considerable interest and might indicate a genetic influence in its causation.


Pseudoxanthoma elasticum and Elastosis perforans serpiginosa. Report of two cases. Two cases of pseudoxanthoma elasticum with elastosis perforans serpiginosa are presented. The two conditions and their association are briefly discussed.


Screening for 6- phosphogluconate dehydrogenase abnormalities in the blood of dermatological patients. The results of screening for 6-phosphogluconate dehydrogenase abnormalities in the blood of dermatological patients are reported. No differences were found between controls, psoriatics, psoriatics receiving methotrexate and a group of mixed patients. The results are discussed in relation to the effects of methotrexate on glucose-6-phosphate dehydrogenase.


Lack of effect of vitamin A on differentiation in squamous carcinoma. High oral doses of vitamin A given to Irish rats bearing squamous carcinomas failed to produce detectable light level histologic change in the tumors.  The cell line was grown in tissue culture with and without vitamin A; electron-microscopic examination failed to reveal significant differences between experimental and control cultures.


[Porphyria cutanea tarda-like aspects in two prolonged hemodialysis patients (author's transl)]. In two female patients with prolonged maintenance hemodialysis, blisters and postbullous erosions could be observed. One case also showed raised amounts of uroporphyrin I and III in the urine.


Pre-ovulatory elevation of rat ovarian prostaglandins F, and its blockade by indomethacin. Prostaglandins of the F series have been measured, by radioimmunoassay, in ovaries of prepubertal rats in which first estrus was synchronized by injection of pregnant mare serum gonadotropin. Marked elevations, up to 20-fold, were observed approximately in parallel with the endogenous luteinizing hormone surge which occurs on the evening of proestrus, and within 4 h after injection of exogenous luteinizing hormone on the morning of proestrus. Injection of indomethacin, 0.8 mg per rat, at 14:45 on proestrus, prevented the increase in ovarian prostaglandins F without significantly reducing the proestrous elevation of serum luteinizing hormone. These observations support elevation of serum luteinizing hormone. These observations support a physiologic role of prostaglandins F in rat ovarian responses to luteinizing hormone.


Studies on the synthesis of estrogen-specific uterine proteins. Comparison of methods of quantitative evaluation of double-isotope peaks. Double-isotope techniques are frequently used to provide a sensitive means of detecting specific biosynthetic responses to experimental manipulations, usually protein or nucleic acid synthesis. The qualtative basis for the detection of changes in biosynthetic rates in these double-isotope experiments involves measuring changes in the ratio of the isotopes to each other after fractionation of the albeled extract. Changes in the isotope ratios have been used directly in quantitative interpretations also, but such quantitative interpretations of changes in isotope ratios are potentially erroneous since the magnitude of the change in ratio of one isotope to the other will depend upon the total number of counts within the area in question (the 'baseline'). The results of experiments on an estrogen-specific uterine protein are presented in this report. It is demonstrated that under conditions in which the 'baseline' incorporation of isotope varies within an experiment, such as in purification or pulse-chase studies, the use of isotope ratios to quantify the data results in erroneous values. In these instances computations using the delta 14C method described by Mayol and Sinsheimer (1970) provides for what appear to be valid conclusions. However, under conditions in which the baseline incorporation of isotope of the control and experimental groups does not vary, as in studies of the temporal course of synthesis and dose-response relationships, the use of increases in isotope ratios is adequate for quantitative interpretations.


Renal adenylate cyclase activation by amino acylated vasopressin and oxytocin. Two series of neurohypophysial peptide amino-acylated derivatives were tested for their ability to activate plasma membrane adenylate cyclase prepared from pig or rat kidney. They were firstly [8-lysine]-vasopressin-related derivatives (Na-[Glycyl-Cys]1-[8-Lysine]-vasopressin and Na-[Glycyl-Glycyl-Cys51-[8-Lysine]-vasopressin) and secondly oxytocin-related derivatives (Na-[Glycyl-Cys-a1)-oxytocin, Na-[Leucyl-Glycyl-Glycyl--Cys]-oxytocin, and Na-[Glycyl-Cys]-[2-0methyl tyrosine]-oxtocin). The maximal adenylate cyclase activation induced by these peptides was lower than that induced by their respective parent hormones. After incubation of these analogues with plasma membranes obtained from the renal medulla, no significant release of parent hormones occurred. Good qualitative correlations were observed between relative antidiuretic activities measured in vivo and relative potencies in activating adenylate cyclase. It was concluded that direct action of peptides tested on the kidney is at least partly responsible for their antidiuretic activity in vivo.


Activation of rat kidney adenylate cyclase by vasopressin analogues: lack of correlation with antidiuretic activity. Vasopressin analogues with enhanced antidiuretic activity in vivo (deamino-[D-arg8]-vasopressin, deamino-6-carba-[Orn8]-vasopressin, deamino-6-carba-[Arg8]-vasopressin, and deamino-6-carba-[D-Arg8]-vasopressin) were tested for their ability to activate rat renal medullary adenylate cyclase and compared to the natural antidiuretic hormones [Arg8]- and [Lys8]-vasopressin. The enzyme preparation used did not inactivate the vasopressins or the analogues tested. The analogues activated adenylate cyclase. However, several of them were far less effective than expected on the basis of their very high in vivo antidiuretic activity. It was concluded that the enhanced in vivo activity reflects greater metabolic stability in vivo rather than enhanced affinity for the renal antidiuretic hormone receptor.


Estradiol bending in cytosol from epididymides of immature rabbits. A highly specific, high affinity binding protein for estradiol-17beta (E2) is present in cytosol prepared from the epididymides of immature (21-53 day old) rabbits. This binding moiety sediments on sucrose gradients as an 8S species under low ionic strength conditions and as a 4S species under conditions of high ionic strength (0.3 M KCL). The relative binding affinities of estrogens for the binding protein was E2 is greater than estrone is greater than estriol. Neither 5alpha-dihydrostestosterone (5alphaKHT), progesterone, nor cortisol were able to inhibit binding of [3H]E2 to epididymal binding sites. An 8S binding moiety for E2 was present in testicular cytosol but not in muscle. An apparently non-specific binding component for E2 was present in plasma which sedimented in the 4S region of low ionic strength gradients. The epididymal E2 binding moiety was distinct from a 4S androen binding protein of testicular origin which is detectable in cytosol prepared from epididymides of rabbits at certain stages development. We were unalbe to detect a specific E2 binding protein in epididymal cytosol from mature intact or 4-day castrated rabbits. The E2 binding component in the cytosol of immature rabbits had an Kd congruent to 2-10 X 10-10 M and the concentration of binding sites was in the order of 1-4 X 10-13 mmoles/mg of protein. The binding component was thermo-labele and pronase, but not nuclease, sensitive.


Regulation of casein synthesis in the rabbit mammary gland.  Titration of mRNA activity for casein under prolactin and progesterone treatments. Total polysomal RNA or poly(A)-containing RNA isolated from membrane-bound polysomes of normal lactating rabbits directed the synthesis of casein in a reticulocyte lysate. Casein was identified by immunoprecipitation followed by SDS-polyacrylamide gel electrophoresis of the immunoprecipitate. The poly(A)-containing RNA was heterogeneous with one major peak corresponding to a 12-S sedimentation coefficient as determined by polyacrylamide gel electrophoresis. Using the same procedure, mRNAs isolated from the non-secreting tissue of pseudopregnant rabbits were found not to contain the 12-S peak and were unable to direct the synthesis of casein in vitro. Prolactin injected into pseudopregnant rabbits induced the synthesis of proteins immunoprecipitable by anti-casein anti-serum and induced the simultaneous appearance of the 12-S mRNA. Progesterone injected with prolactin prevented the induction of casein synthesis and the appearance of mRNA for casein. A close relationship was established between the ability of the tissue to synthesize immunoprecipitable casein and the corresponding mRNA content of polysomes.


The purity of allergenic extracts.

The purity of allergenic extracts. The purity of allergenic extracts is usually taken for granted but we know that trace amounts of impurities may be the most important determinant causing clinical symptoms. We know that many commercial extracts are grossly contaminated with other allergens and this is easy to demonstrate by skin tests. Horse or dog scurf collected in the summer months will be contaminated by pollens. Patients can be shown to have organ specific inhalant allergic complaints due to one animal. Sometimes the allergenicity is at first only towards an antigen in the saliva, then skin antigen(s) and finally all organs of the body may cause allergic symptoms, so that urinary protein as well as serum albumin cause reactions to the patients. When collecting individual pollens, it is very difficult not to get these contaminated by other pollens. Named grass pollens will be contaminated by other grass pollens and fungal spores which are airborne at the same time. An extract which has been labelled plantain pollen will often be contaminated by 1% or more of grass pollen. A tree pollen such as Tilia is always grossly contaminated with other pollens. A mite extract has in it fish, human serum albumin from skin scales or horse, depending on the culture media used for feeding the mites. Animal scurf extracts are nearly always contaminated with mites. We can conclude that allergenic extracts are difficult to produce immunologically pure and they are often grossly contaminated.


Stability of antigen E in commercially prepared ragweed pollen extracts. Aqueous ragweed pollen extracts (20,000 PNU/ml or 78,000 PNU/ml) lose 50% of their Antigen E content within 5 months of storage at 4 degrees C. Glycerinated extracts and lyophilized ragweed pollen extracts retain their Antigen E content for over 15 months with little or no loss. The preservative phenol in aqueous extracts is destructive to Antigen E probably by protein denaturation. The stability of ragweed extracts as measured by Antigen E content is markedly enhanced with little loss after 15 month storage at 4 degrees C, when the extract is prepared as an alum precipitated suspension.


Some considerations on the production and standardization of bacterial allergens. The production and standardization of the bacterial allergens are very difficult and connected with many unknown and disputable problems. The authors have carried out a study with three basic bacterial allergens produced by themselves: bacterial extracts, bacterial suspensions and a mixture of both from Streptococcus hemolyticus and Staphylococcus albus. The specific activity of all allergens in different concentrations is determined by skin intradermal tests simultaneously on 22 patients sensitized to Str. hemolyticus. 23 patients sensitized to Staph. albus, as well as on 10 healthy persons. The results of the study show that the bacterial extracts give predominantly immediate type skin allergic reaction while the bacterial suspensions lead to delayed type reactions. The mixed allergens give both type skin reactions, and reveal more specifically sensitization against the bacteria. The authors suggest that the optimal concentration of the mixed bacterial allergens for diagnosis by intradermal test is 10,000 PNU/ml and 100,000,000 bacterial/ml. The advantages and disadvantages of the three bacterial allergens are discussed.


[Preparation and standarization of a Brucellosis allergen of sheep and goats]. The authors underline the consequences upon national economy and public health of sheep and goat brucellosis and expose the difficulties existing with the serological methods of diagnosis presently in use. In order to eliminate these disadvantages they propose the use of the intradermal allergic test reporting, at the same time, the reasons for which this test has not been until now universally adopted : lack of a standard allergen, always prepared by the same method and possessing excellent qualities. To avoid existing difficulties, they propose the use of a new allergen based on an original method of preparation, resulting from a careful study of the subject. They describe the method of preparation of the product and expose the different laboratory control tests on albinos guinea pigs. The excellent results obtained allow to conclude that the brucella melitensis strain used is suitable for the preparation of the allergen as well as for the sensitization of guinea pigs. The best sensitization method is the double subcutaneous injection of viable organisms in emulsion with Freund's adjuvant : one billion for the first and two billions for the second injection, at twenty days interval. Activity control of the allergen by intradermal inoculation of dilutions 1/10, 1/1001/1000, fifteen days after the last injection, gives the best results. Reading of the allergic reactions should take place twenty-four hours after the intradermal inoculation of the allergen because of rapid decrease in dimensions and intensity. The allergen demonstrates exceptional potency and specificity qualities; it may be considered as a "purified" product : all the tests on the control guinea pigs are negative, there is no formation of agglutinins nor sensitization of these laboratory animals towards the product itself.


RAST-based allergen assay methods.

RAST-based allergen assay methods. The RAST principle has been utilized for the detection and assay of allergens of various origins. RAST-based methods have the advantage of being convenient, accurate and specific by making use of the IgE antibodies, considered as mediators of the atopic allergic reactions, as reagents in vitro. Two improved assay procedures, a direct titration and an inhibition method, have been used for the estimation of extract potencies and for the assay of separate allergens. The specificity of the assays is determined by the specificity of the IgE (sera from allergic patients) used as the main reagent. Specific assay of separate components in heterogeneous extracts may be achieved in some cases by careful characterization of the sera or by selective blocking of irrelevant IgE. The potency of allergen concentration may be calculated and expressed in arbitrary units, and the accuracy and sensitivity of the two methods are similar. The coefficient of variation is in the range 10 to 25% provided that standardized reagents are used. Sensitivity of the assays is about 200 ng per ml. The lower limit for detection of allergen is 10-100 times lower.


Grass allergens : a preliminary characterization by means of RAST-based allergen assay methods. A preliminary study of a grass pollen allergens by means of radioallergosorbent test (RAST)-based assay methods has been made. A combination of gel filtration and ion exchange chromatography has been used to separate allergens, mainly from Festuca elatior, Lolium perenne, and Phleum pratense, into groups dependent on their molecular weight and net electric charges. A RAST-based titration method was used to detect and quantitate the different allergens. The specificity towards IgE of the different allergen groups was investigated by means of a RAST-based inhibition method. The allergen groups were found to be of a quite heterogenous nature when investigated by means of polyacrylamide gel electrophoreses. Furthermore, the different groups exhibited a considerable degree of cross-reactions when tested against a pool of patient sera by means of the inhibition method. This was the case even for allergen groups which differed significantly both in regard to molecular weight and net electric charge.


Studies on antigens from timothy pollen. A variety of dialyzable and non-dialyzable fractions from neutral timothy pollen extracts were studied in human and guinea pig RAST inhibition as well as in PCA. Extracted material fragmented by chemical reagents and by hydrolytic enzymes was also included in the study. It was found that some low molecular weight fractions were quite active in PCA and in RAST inhibition. The fragmented materials showed in a number of instances a considerably reduced inhibitory capacity in RAST but remarkably high PCA elicitation. Some of the enzymes did not produce significant modifications detectable by RAST inhibition although the test seems to be very sensitive to antigenic modification. It seems quite certain that a considerable number of antigenic specificities are involved in the clinical sera tested.


A new method of measuring histamine within single cells using a microspectrophotofluorimeter. Methods of measuring histamine from large numbers of cells involve inaccuracies due to the difficulty of isolating a pure cell population without damage to the cells. A new method of measuring histamine within single cells by a microspectrophotofluorimetric method has been developed. Calibration of the microspectrophotofluorimeter used o-phtalaldehyde-conjugated-histamine absorbed to Sephadex particles. The concentration of histamine within single rat mast cells was measured and compared with the theoretical values obtained from measuring histamine released from large cell samples.


Standardization of stinging insect extracts. The measurement of IgE Ab to insects (bee etc.) can be accomplished using RAST-discs. This prevents the non-specific wheal and flare produced when skin tests in higher concentrations of either whole body or venom are used. It will also eliminate the possibility of a serious systemic reaction by direct skin testing. The in vitro neutralization of IgE antibody by pure fractions and or components employing the RAST disc method may well be a useful tool for standardization of allergens.


Standardization of bacterial allergens.

Standardization of bacterial allergens. One of the prerequisites for the standardization of bacterial allergens consists in the development and formulation of uniform requirements for the assessment of the specific activity of allergens, taking into account the problems to be solved by means of these allergens: a) the screening of large groups of population in order to select those that should be vaccinated or b) the evaluation of the extent of allergic conversion in humans. The data presented here have been obtained in the Laboratory of Allergens of L.A. Tarassevich State Research Institute for Standardization and Control of Medical Biological Preparations. They show that the frequency of skin reactions depends on the concentration of protein in a single dose (e.g. with hemolytic streptococcus allergen) as well as on the properties of the bacterial strain (e.g. with staphylococcal allergens). The standardization of allergens involves the use of special in vitro reactions of blood leukocytes. In 1962 Fradkin proposed the NDI (neutrophile damage index) test based on the enhancement of the amoeboid activity of neutrophiles after the incubation of blood smears to which allergen has been added. The results of differentiation of infections and postvaccinal allergy in children by means of NDI test are discussed. It is suggested that specific forms of allergens should be manufactured for in vitro cellular reactions.


Partial characterization of human histaminopectic factor. Sera from normal human subjects are pooled and fractions are isolated by salt precipitation. The fraction responsible for agglutinating histamine bound latex particles is further purified by column chromatography. The histaminopectic activity can thus be traced to a single protein present in the IgG fraction of normal human serum. Its absence or the presence of an inhibitor in serum from allergic subjects is also sought as well as correlations between plasma concentrations of known immunoglobulins and specific histaminopectic activity. Some of its physical-chemical properties are explored. Evidence was also presented pointing to the possible presence of an inhibitor of the histaminopectic factor in human atopic serum.


Examination of cocksfoot pollen (Dactylis glomerata) extracts by skin prick test in man, RAST-based allergen assay and protein content. Three commercial preparations of cocksfoot pollen (Dactylis glomerata) from different manufacturers were compared by skin prick tests in man, allergen activity as measured by the RAST test and protein nitrogen unit (PNU) and total nitrogen (N) content. The skin prick tests were made on the forearm with the undiluted preparations and also with solutions of 1/5 and 1/25 dilutions. Three groups each of 9 subjects were tested and the sites for the 9 tests were determined by 3 different latin squares. Taking one of the preparations as having a potency of unity, the estimated potencies for the other two were 0.28 and 0.18 when the areas of the weals were measured by a graph paper method and they were 0.27 and 0.19 when the areas were calculated by taking the product of the longest diameter of the weal and that at right angles to it. No notable differences were observed when the solutions were inoculated at different test sites. The three preparations were ranked in the same order of potency by the skin prick test, by measurement of allergen activity as measured by a RAST-based allergen assay method using the sera of the test subjects, and by measurement of their PNU but not their total N content. Comparisons such as these could possibly lead to the production of allergen extract preparations of similar or with consistent potency.


[Allergens adsorbed on aluminum hydroxide preparation, standardization controls and therapeutic results]. Specific desensitizing treatments with house dust extract purified and adsorbed on aluminium hydroxyde gave 79% good and very good results in 355 patients. Grass pollen extracts adsorbed on the same adjuvant gave 76% good and very good results in 166 patients. Desensitization by the adsorbed preparations requires fewer injections than the usual methods, while offering security by the absence of anaphylactic reactions, particularly with pollen extracts. The activity of the preparations is tested after appropriate dilution, taking account of the total nitrogen content of the dust extracts and the weight of raw materials used in the manufacture of the pollen extracts. The results of the intradermal tests undertaken with fluid preparations and compared with those obtained with a standard preparation allow to standardize the allergenic activity of the extracts. It is thus possible to guarantee the pharmaceutical quality of the allergens both for diagnostic tests and for treatments. Chemical analyses and control of acute, delayed or chronic toxicity ensure the innocuousness of the preparations.


Standardization of allergen extracts.

Standardization of allergen extracts. The biological activity in the allergic patient is our first and most relevant concern. The physician using an extract must know the quality and degree of allergenic activity in the material. Preferably, it should also be possible for the physician to control such activity by simple techniques. No single test can fully satisfy these demands. The standardization which comes nearest to this is assessment of the biological activity and specificity by means of skin testing using histamine as a reference. Skin testing is, however, limited as regards precision and reproducibility. A more precise standardization may be achieved by means of certain in vitro methods. Relevant in vitro methods are suitable complements to the bio-assay. However, the in vitro methods chosen must be standardized themselves as compared to a stable reference, and their correlation with the biological reactions in man must be established before they can be introduced as possible means for extract standardization. Even then, bio-assays are necessary for control of content of irritants which may elicit non-immunological (non-specific) reactions in the hyperreactive tissues of allergic individuals. Methods for standardization are discussed. Examples of quality gradings of allergen extracts are given with emphasis on minimal requirements to be fulfilled by extracts in order to be accepted for clinical use.


Profiles of allergen extract components by isoelectric focussing and radioimmunoassay. When allergen extracts are subjected to isoelectric focussing in polyacrylamide gel, followed by radioimmunoassay different preparations of nominally the same allergen show markedly different concentrations of components. Not all the peaks are likely to be due to components of different antigenic specificity, but undoubtedly the sera from different patients contain different spectra of antibodies to the same preparation.


A radioimmunoassay for chemically modified grass pollen allergen. Because of the physicochemical properties of tyrosine adsorbed chemically modified grass pollen allergens neither the human skin test in atopic volunteers nor the inhibition of the radioallergosorbent test are suitable assay procedures. Radioimmunoassays involving a labelled IgG fraction of rabbit antisera to the modified materials can be developed. The dilution inherent in the solubilisation procedure for the tyrosine does not, in most cases, take the antigen concentration below the level of sensitivity of the assay. Assay of different batches of Pollinex with an inhibition assay shows that discernment of different dose levels is possible. The results are compared with a method of assaying total protein.


Standardization of housedust allergen in human skin test trials. Standardization was carried out by comparison of Bencard and our own ('Human') housedust allergens in human skin test trials. The dose response curves on hypersensitive patients were parallel, linear, and there was no difference between preparations according to the variance analysis. The relative potency was 1.33 compared to the Bencard allergen. 95% confidence limits were 0.29-2.37. The correlation coefficient evaluated from all data was 0.90. Our experience suggests that human skin test trials seem to be a simple and convenient method for the standardization of housedust allergens. According to the results of analysis of factorials, data suitable for standardization can be obtained if the area of both the wheal and flare as different parameters are recorded 10 and/or 20 minutes after the intradermal injections.


Physico-chemical properties and specific activity of a purified allergen (codfish). The major allergen (Allergen M) from codfish is an acidic protein, mol. wt. 15000 with 114 amino acid residues and only one sugar (glucose) residue. It is highly active in skin testing on codfish-allergic individuals, in passive transfer (PK) testing and in the radioallergosorbent test (RAST). The activity resists denaturation and incomplete tryptic hydrolysis. Two fragments (TM 1, mol. wt. 8500, and TM 2, mol. wt. 6500) are obtained from allergen M. The fragments are equally active and allergenically identical in biological tests, but somewhat less active than Allergen M itself. Smaller peptides obtained from TM 2 are inactive. The amino acid sequence of TM 2 is given as the first reported example of a full structural analysis of an active allergen. The information obtained in studies of this model allergen is compared with information published from other studies of allergens. Similarities and differences are pointed out.


Antifibrin action of phenformin.

Antifibrin action of phenformin. Effects of phenformin on blood sugar, serum triglyceride, thrombin time, euglobulin clot lysis time and cardiovascular complications were studied in maturity onset diabetes and in atherosclerotic patients with or without diabetes, for a period of 14-18 months. Phenformin has shown the characteristic properties of an antifibrinopathic agent in that it prolongs thrombin time and enhances fibrinolysis. The hypoglycaemic effect of phenformin was found to be directly related to its antifibrinopathic action. Plasma lipids fell in all cases. Absence of fresh cardiovascular complications and improvement in anginal symptoms were observed. The metabolic, haematological and clinical benefits of phenformin and its limitations in maturity onset diabetes and atherosclerosis may be explained by the effects of the drug upon the thrombin-fibrinogen reaction. These results lend support to the hypothesis of a primary fibrinopathic pathogenesis in maturity onset diabetes mellitus and atherosclerosis.


Inhibition of bile salt absorption by blood-sugar lowering biguanides. The effect of blood sugar lowering biguanides (phenethyl-, butyl- and dimethylbiguanide) upon jejunal and ileal transport of bile salts (tauro- and glycocholate) was tested in rat small intestine by an in vitro technique. Giguanides inhibited active transport of bile salts in the ileum, but did not affect diffusional absorption of bile salts in the jejunum. The inhibitory effect was time-dependent and not reversible under in vitro incubation conditions, suggesting that biguanides must enter intestinal mucosal cells in order to exert their inhibitory action on active transport of glucose analogues, amino acids, calcium and bile salts. Since biguanides achieve high tissue concentrations in the small intestine even after parenteral administration, inhibition of ileal bile salt reabsorption by biguanides could possibly explain the lipid- and cholesterol-lowering effect of these oral antidiabetic drugs.


The effect of phenformin on amino acid-induced insulin secretion in diabetics. Twenty-one patients with mild maturity-onset diabetes were given introduodenal infusions of an amino acid mixture (0.5 g amino acids per kg body weight). In 9 other patients L-arginine was infused intravenously in a constant dose of 25 g. Alpha-amino nitrogen, blood glucose and plasma insulin levels were assayed under control conditions and after three days of treatment with phenformin, 150 mg daily, plus the same 150 mg dose 60 min before the second loading. Intraduodenal infusion of the amino acid mixture provoked a greater increase in plasma insulin than intravenous infusion of L-arginine, this increase being significantly inhibited by phenformin only in the first case. Since no evident influence of phenformin on the intestinal absorption of amino acids could be demonstrated, this effect may be explained by a local action on the intestinal wall exposed to high concentrations of the drug, resulting in the inhibition of the insulin secretion stimulating activity of the gut.


The incidence of diabetes mellitus in the offspring of diabetic couples. Investigation based on the oral glucose tolerance test. 88 of 137 offspring of 22 adult onset diabetic couples were subjected to an oral glucose tolerance test (G.T.T.); 35 were found to have a diabetic G.T.T. As 8 were already known as diabetics, 43 of 96 offspring (45%) were found to have diabetes mellitus. Neither the data pertaining to the parents, such as family history, treatment of diabetes or obstetric history, nor the data pertaining to the offspring, such as sex, age, parity or course of pregnancy, could be related to the incidence of diabetes in the offspring. It was noted however, that overweight female, but not male, offspring are more likely to develop diabetes than those who are not overweight (p less than 0.05).


Serum IRI in insulin-treated diabetics during a 24-hour period. Serum immunoreactive insulin extracted with acid ethanol (total IRI) and blood glucose were measured in two groups of diabetics and a control group during 24-hour periods. One group of diabetics had received insulin for less than 1 month, and none had yet developed insulin antibodies. The other had been treated with insulin for 2 years or more. The average level of total IRI in the diabetics without antibodies was similar to that in the normals, but highly elevated in diabetics with antibodies, although the blood glucose levels were similar in the two diabetic groups. The increase in total IRI after the insulin injections was more rapid in patients receiving quick-acting insulin and most pronounced in those with the highest levels of total IRI. The total IRI peaked in the afternoon and dropped during the night. The short-term insulin-treated patients showed a better degree of diabetes control than those treated for 2 years or more. No correlation was found however, between the total IRI level and the degree of control in the latter group. The patients with the highest levels of total IRI had the highest frequency of hypoglycemic episodes. This is in agreement with the concept that the insulin antibodies may function as a circulating depot, which releases insulin irrespective of the metabolic need.


Cell size and the antilipolytic effect of insulin in human subcutaneous adipose tissue. Lipolysis was studied in subcutaneous adipose tissue removed under local or general anaesthesia from subjects with marked difference in body weight and from obese subjects before and after intestinal shunt operations. The release of glycerol was measured in sections of adipose tissue incubated for two hours in bicarbonate buffer containing 4 per cent bovine albumin with no glucose added. The larger fat cells were more insulin sensitive than the smaller. A positive relationship was observed between the fat cell size and the antilipolytic effect of insulin (100 muU/ml). Insulin had no significant effect on the lipolytic effect of the catecholamines, when this is calculated as the increment in the glycerol release above the basal.