Extra-vesicular binding of noradrenaline and guanethidine in the adrenergic neurones of the rat heart: a proposed site of action of adrenergic neurone blocking agents. 1 The binding and efflux characteristics of [14C]-guanethidine and [3H]-noradrenaline were studied in heart slices from rats which were pretreated with reserpine and nialamide. 2 Binding of both compounds occurred at extra-vesicular sites within the adrenergic neurone. After a brief period of rapid washout, the efflux of [14C]-guanethidine and [3H]-noradrenaline proceeded at a steady rate. The efflux of both compounds appeared to occur from a single intraneuronal compartment. 3 (+)-Amphetamine accelerated the efflux of [14C]-noradrenaline; this effect was inhibited by desipramine. 4 Unlabelled guanethidine and amantadine also increased the efflux of labelled compounds. Cocaine in high concentrations increased slightly the efflux of [14C]-guanethidine but not that of [3H]-noradrenaline. 5 Heart slices labelled with [3H]-noradrenaline became refractory to successive exposures to releasing agents although an appreciable amount of labelled compound was still present in in these slices. 6 It is suggested that [14C]-guanethidine and [3H]-noradrenaline are bound at a common extravesicular site within the adrenergic neurone. Binding of guanethidine to the extra-vesicular site may be relevant to its pharmacological action, i.e., the blockade of adrenergic transmission.


Effect of morphine and naloxone on priming-induced audiogenic seizures in BALB/c mice. 1 Morphine (1-200 mg/kg s.c.) reduced the incidence and prolonged the latency of priming-induced audiogenic siezures in a dose-dependent manner. 2 This effect was reversed by naloxone (1 and 2 mg/kg) although naloxone was itself inactive. 3 This priming-induces seizure model may be useful in the study of tolerance and physical dependence.


Long-term effects of N-2-chlorethyl-N-ethyl-2-bromobenzylamine hydrochloride on noradrenergic neurones in the rat brain and heart. 1 N-2-Chlorethyl-N-ethyl-2-bromobenzylamine hydrochloride (DSP 4) 50 mg/kg intraperitoneally, produced a long-term decrease in the capacity of brain homogenates to accumulate noradrenaline with significant effect 8 months after the injection. It had no effect on the noradrenaline uptake in homogenates from the striatum (dopamine neurones) and on the uptake of 5-hydroxytryptamine (5-HT) in various brain regions. 2 In vitro DSP 4 inhibited the noradrenaline uptake in a cortical homogenate with an IC50 value of 2 muM but was more than ten times less active on the dopamine uptake in a striatal homogenate and the 5-HT uptake in a cortical homogenate. 3 DSP 4 (50 mg/kg i.p.) inhibited the uptake of noradrenaline in the rat heart atrium in vitro but this action was terminated within 2 weeks. 4 DSP 4 (50 mg/kg i.p.) cuased a decrease in the dopamine-beta-hydroxylase (DBH) activity in the rat brain and heart. The onset of this effect was slow; in heart a lag period of 2-4 days was noted. In brain the DBH-activity in cerebral cortex was much more decreased than that in hypothalamus which was only slightly affected. A significant effect was still found 8 months after the injection. The noradrenaline concentration in the brain was greatly decreased for at least two weeks, whereas noradrenaline in heart was only temporarily reduced. 5 The long-term effects of DSP 4 on the noradrenaline accumulation, the DBH activity and noradrenaline concentration in the rat brain were antagonized by desipramine (10 mg/kg i.p.). 6 It is suggested that DSP 4 primarily attacks the membranal noradrenaline uptake sites forming a covalent bond and that the nerve terminals, as a result of this binding, degenerate.


Inhibition by amiloride of sodium-dependent fluid reabsorption in the rat isolated caudal epididymis. The rate of fluid reabsorption was studied in the rat isolated caudal epididymal sac in vitro. 2 Part of the fluid reabsorption was found to be dependent on intraluminal Na+. Amiloride (0.1 mM) completely inhibited this component of fluid reabsorption. 3 The log dose-inhibition curve to amiloride was sigmoid and the IC50 value was found to be 1.6 muM.


Myocardial uptake of digoxin in chronically digitalized dogs. 1 The time course of myocardial uptake of digoxin, increase in contractility and changes in myocardial potassium concentration was studied for 90 min following an intravenous digoxin dose to long-term digitalized dogs. 2 Nineteen dogs were investigated by the use of a biopsy technique which allowed sampling before and after administration of digoxin. 3 Ten minutes after administration of digoxin the myocardial concentration increased from 60 to 306 nmol/kg tissue, the myocardial concentration of digoxin was significantly lower (250 nmol/kg tissue) after 30 min and then increased again. 4 The transmural myocardial distribution of digoxin was uniform before and 90 min after administration of digoxin in long-term digitalized dogs but at 10 min after administration, both the subepicardial and the subendocardial concentration of digoxin were significantly lower than that of the mesocardial layer. 5 During the first 10 min the dp/dtmax increased to 135% of the control level. The increase remained unchanged during the rest of the study. 6 Myocardial potassium decreased throughout the study. 7 The M-configuration of the myocardial uptake curve and the non-uniformity of myocardial distribution of digoxin observed at 10 min after administrating digoxin to long-term digitalized dogs indicate that the distribution of myocardial blood flow may be changed during chronic digitalization.


Studies on the mechanism of action of amantadine. 1 The effect of amantadine hydrochloride on various aspects of catecholamine metabolism in the rat brain has been investigated. 2 Amantadine failed to have any significant effect on brain concentrations of dopamine or noradrenaline even when administered daily for 9 days. 3 Amantadine had no effect on the rate of decline of noradrenaline and dopamine concentrations after alpha-methyl-p-tyrosine. 4 In vitro amantadine inhibited dopamine uptake into synaptosomes only at high concentrations, and caused little release of dopamine from synaptosomes. 5 There is no evidence from these results to suggest that the anti-Parkinsonian effect of amantadine is related to an action on dopaminergic mechanisms.


Further evidence that prostaglandins inhibit the release of noradrenaline from adrenergic nerve terminals by restriction of availability of calcium. 1 Guinea-pig vasa deferentia were continuously superfused after labelling the transmitter stores with [3H](-)-noradrenaline. Release of [3H]-(-)-noradrenaline was induced by transmural nerve stimulation. 2 Prostglandin E2 (14 nM) drastically reduced the release of [3H]-(-)-noradrenaline, while tetraethylammonium (2 mM), rubidium (6 mM), phenoxybenzamine (3 muM) each in the presence or absence of Uptake 1 or 2 blockade, and prolonged pulse duration (from 0.5 to 2.0 ms) all significantly increased the release of [3H]-(-)-noradrenaline per nerve impulse. 3 The inhibitory effect of prostaglandin E2 on evoked release of [3H]-(-)-noradrenaline was significantly reduced by tetraethylammonium, rubidium and prolonged pulse duration, whilst it was actually enhanced by phenoxybenzamine. This indicates that increased release of noradrenaline per nerve impulse does not per se counteract the inhibitory effect of prostaglandin E2. 4 It is concluded that tetraethylammonium, rubidium and prolonged pulse duration counteracted the inhibitory effect of prostaglandin E2 on T3H]-(-)-noradrenaline release by promoting calcium influx during the nerve action potential. The results are consistent with, and add more weight to the view that prostaglandins inhibit the release of noradrenaline by restriction of calcium availability.


A comparison of affinity constants for muscarine-sensitive acetylcholine receptors in guinea-pig atrial pacemaker cells at 29 degrees C and in ileum at 29 degrees C and 37 degrees C. 1 The affinity of 17 compounds for muscarine-sensitive acetylcholine receptors in atrial pacemaker cells and ileum of the guinea-pig has been measured at 29 degrees C in Ringer-Locke solution. Measurements were also made at 37 degrees C with 7 of them. 2 Some of the compounds had much higher affinity for the receptors in the ileum than for those in the atria. For the most selective compound, 4-diphenylacetoxy-N-methylpiperidine methiodide, the difference was approximately 20-fold. The receptors in the atria are therefore different the structure from those in the ileum. 3 The effect of temperature on affinity are not the same for all the compounds, tested indicating different enthalpies and entropies of adsorption and accounting for some of the difficulty experienced in predicting the affinity of new compounds.


A comparison of deaf and non-deaf patients with paranoid and affective psychoses. A comparison was made of the social and domestic background, prepsychotic personality and symptomatology of deaf ('hard of hearing') and non-deaf patients aged 50 or over with paranoid or affective psychoses. The deaf patients were found to be a very heterogeneous group in respect of age of onset, duration, severity and pathological basis of their deafness, but there was a substantial subgroup of paranoid patients with deafness beginning before the age of 45 in whom the personality appeared to have been less deviating than in the remainder. It was thought that the deafness in this subgroup had possibly played a relatively specific role in causing the psychosis. Some problems in identifying deafness and assessing its significance are discussed.


A psychosocial study of accidental poisoning in Egyptian children. One hundred and twenty-seven accidentally poisoned children from the emergency room of the Islamic Girls College, Cairo, were compared with an equal number of controls. The highest age incidence for both sexes in poisoned children was 36 months. Males outnumbered females at a ratio of 3:2. Behavioural problems such as hyperactivity, temper tantrums, aggression, stubbornness, nocturnal enuresis and impulsiveness occurred more often in poisoned children than in controls and more often in those (24 patients) referred with accidental poisoning on more than one occasion. The families of poisoned children differed significantly from the controls in their large size, low level of education, disturbed home atmosphere and the accessibility to the child of the poisonous substance.


Recruitment of problem drinkers.

Recruitment of problem drinkers. As the first phase of a project designed to institute controlled drinking as a therapeutic goal for alcoholics, it was decied to recruit volunteers who were dissatisfied with their present mode of drinking but who had not previously sought help. This was done by requesting them to phone a private number. When these volunteers had been recruited it was proposed to modify their drinking habits to their own requirements, using non-averse training, drinking practice and ongoing group psychotherapy. This project however has not been implemented because of paucity of volunteers. The methods of recruitement are listed and those drinkers who volunteered are described. The reasons for the low rate of response are discussed.


Clozapine versus perphenazine: the value of the biochemical mode of action of neuroleptics in predicting their therapeutic activity. The chemical structure of a neuroleptic does not relaibly predict the exact profile of its therapeutic action. We considered the question whether the biochemical action of a neuroleptic, and specifically the ratio between DA-receptor block and NA-receptor block, might have a higher predictive value in this respect. In this context we carried out a double-blind study of the therapeutic value of clozapine and perphenazine in acute psychoses of varying symptomatology anc aetiology. There are strong indications that clozapine has only a slight inhibitory effect on transmission in central DA-ergic neurons, but markedly inhibits transmission in central NA-ergic neurons, and that the reverse applies to perphenazine. In view of these data we expected perphenazine to be a stronger antipsychotic and a weaker sedative than clozapine, and vice versa. The plausibility of this hypothesis was demonstrated. Partly also on the basis of earlier research, we concluded that the biochemical action of a neuroleptic is a more faithful predictor of its therapeutic action profile than the chemical structure.


Is a homosexual orientation irreversible? In four studies homosexual patients were randomly allocated to receive either different forms of aversive therapy; or aversive therapy or a positive conditioning procedure. Before and after therapy the patients' sexual orientation was determined by measuring their penile volume responses to moving pictures of nude men and women. The aversive therapies produced reduction in homosexual feelings and behaviour. In the fourth study the reduction was significantly greater following the aversive compared with the positive conditioning procedure. No significant change in measured sexual orientation occurred in the patients who received aversive therapy compared with those who received positive conditioning. The measured sexual orientation of the homosexual patients after treatment differed significantly from that of heterosexual patients who received aversive therapy for sexual anomalies.


Height and personality characteristics of 47, XYY males in a sample of tall non-institutionalized males. A sample of 471 enlisted men 183 cm or taller serving in the US Navy, Coast Guard, and Marine Corps was screened for Y-chromosome aneuploidy by use of quinacrine fluorescence of peripheral blood smears. Two 47,XYY males were detected, resulting in a prevalence of 00425 or approximately 1 in 236. The prevalence of 47,XYY males (00331) in a number of samples of tall, non-institutionalized males is significantly higher than the incidence in newborn males (00061), indicating that 47,XYY males are disproportionately represented in tall male populations. The 47,XYY males had significantly higher scores than 46,XY males on the Schizophrenia, Schizophrenia+1K, and Prejudice scales of the Minnesota Multiphasic Personality Inventory and significantly lower scores on the Dominance scale. Since the probability that a randomly selected pair of subjects in the sample had four or more scale scores significantly different from the remainder of the group was greater than 05, it is possible that the differences between the 47,XYY and 46,XY males occurred by chance. On the other hand, one or more of these scales may measure personality dimensions on which non-institutionalized 47,XYY males may, in fact, differ from 46,XY males.


Associative intrusions in the vocabulary of schizophrenic and other patients. Associative intrusions in the thought of short-term and long-term schizophrenic, non-schizophrenic, and non-psychiatric in-patients were measured using Rattan and Chapman's multiple-choice vocabulary test. This test consists of two subtests, one with associative distractors and the other without distractors, which are matched on discriminating power and thus permit the assessment of differential specific performance deficit uncontaminated by the generalized deficit characterisitc of many patient groups. Results indicated that some susceptibility to associative intrusions characterizes all groups of long-term patients regardless of diagnosis. Short-term patients (with the possible exception of schizophrenic) did not show such heightened susceptibility. Lengthy illness and/or stay in hospital are apparently implicated in at least this form of thought disorder, which has been often considered to be a uniquely schizophrenic phenomenon.


Elongation of pause-time in speech: a simple, objective measure of motor retardation in depression. A sample of 'automatic speech' (counting from 1 to 10) was tape-recorded and the times taken up by phonation and pauses measured. In four healthy volunteers, both phonation times and pause times remained constant over a period of two months. In four moderately depressed patients, with no history of manic illness and with no obvious clinical signs of motor retardation, the pause times were significantly elongated while the patients were depressed compared to pause times measured after recovery. The phonation times were constant throughout the period of observation (4-6 months). Other tests for motor retardation (tapping time; Nurses' rating scale; Hamilton Retardation Scores) did not give consistent results. It is concluded that the simple test described here may reveal a degree of motor retardation in cases where other tests fail to do so.


Further observation on study difficulty in university students including 'syllabus-boundess'. The UCLS questionnaire, in a form modified to include a measure of syllabus-boundness, and a questionnaire to measure psychiatric symptomatology (the MHQ) were administered to two groups of students, one seeking help for emotional problems, the other a control group. Groups were compared on tests, test findings were inter-correlated, and scores were related to academic success. The UCLSQ is confirmed as a reliable research instrument. Principal component analysis again indicates a separation of psychoneurotic and motivational components of study difficulty. Syllabus-boundness ('Sylbism') emerges as a relatively independent trait, with a significant negative relationship to work satisfaction in both groups. MHQ scores again show a positive correlation between phobic anxiety and academic attainment for patients.


Group modification of affective verbailzations: resistance to extinction and generalization effects. An operant group procedure significantly increased the frequency of here-and-now affect, feedback, and empathy statements made by two groups of four subjects during ten one-hour sessions, divided into base line, acquisition, extinction, reacquisition and generalization periods. The continously reinforced group showed resistance to extinction, attributed in part to reinforcement by other group members when a subject made reinforceable statements. Failure of the variably reinforced group to replicate this finding was attributed in part to a less adequate conceptual grasp of the reinforcement categories resulting from the reduced feedback associated with a variable ratio schedule. Both groups showed strong generalization effects when subjects were distributed among eight new groups and the new groups and the new subjects' performance was also significantly higher than typical base levels.


A new personal disturbance scale (DSSI/sAD). A brief self-report measure of personal disturbance is presented. Being derived from the Delusions-Symptoms-States Inventory, it focuses exclusively on recent symptomatology, uncontaminated by personality attributes. Data are presented which show significant agreement (a) for the allocation of the items to syndromes by experiences raters, and (b) between patients' self-report and their psychiatrists' ratings. At the anxiety, depression, and total sAD scale levels a high discrimination is found between the normal and pmal distributions, both of which are in contrast to personality measures. The scales appear relevant to treatment evaluation and for detecting the personally disturbed in general populations.


Increasing patients' satisfaction with communications. This paper reports a successful experimental attempt to increase satisfaction with communications amongst general medical patients. A group of patients who received extra interviews designed to increase their understanding of what they had been told about their illnesses showed significantly greater satisfaction than patients treated in the usual manner, and patients who received extra placebo interviews about their hospital stay. Changes in satisfaction with communications over the period following discharge from hospital are also described and possible reasons for them are discussed.


Towards a theory of auditory hallucinations: outline of an hypothetical four-factor model. The research findings of the writer to date on auditorily hallucinated patients are briefly summarized and an hypothetical four-factor model outlined to account for them. This general working model envisages auditory hallucinations as stemming from the interaction of psychological stress with an underlying hallucinatroy predisposition which is to a large extent genetically determined. A third factor of external stimulation is seen as exerting an inhibitory effect on the conscious experience of the hallucination. While a fourth speculative factor of a positive feedback loop is introduced to account for a number of otherwise  incompatible observations. Each of the four factors in the hypothetical model is defined as precisely as is possible at present and discussed in relation to other research findings in this area and to the literature more generally. An attempt is made to spell out some of the theoretical predictions and clinical implications of the model.


Biofeedback control of migraine: a pilot study. A group of six subjects was trained, using biofeedback procedures, in the voluntary regulation of hand temperature as a means of controlling migraine attacks. The training procedure and its relationship to migraine activity is described. Daily records of migraine activity were kept during base-line, training, and follow-up periods. Analysis of this data revealed statistically and clinically significant improvement on several indices of migraine activity. Evidence for a concomitant improvement in the ability of subjects to produce voluntary increases in hand temperature is presented.


A review of the pharmacology, pharmacokinetics and behavioral effects of procaine in thoroughbred horses. Since procaine has both local anaesthetic and central stimulant actions its presence in the blood or urine of racing horses is forbidden. After rapid intravenous injection of procaine HC1 (2.5 mg/Kg) in thoroughbred mares plasma levels of this drug fell rapidly (t 1/2 alpha = 5 min) and then more slowly (t 1/2 beta = 50.2 min). These kinetics were well fitted by a two compartment open model (Model I). This model gave an apparent Vdbeta for procaine in the horse of about 3,500 litres. Since procaine was about 45% bound to equine plasma protein this gives a true Vdbeta for procaine of about 6,500 litres. After subcutaneous injection of procaine HC1 (3.3 mg/Kg) plasma levels peaked at about 400 ng/ml and then declined with a half-life of about 75 minutes. These data were well fitted by Model I when this was modified to include simple first order absorption (K = 0.048 min-1) from the subcutaneous injection site (Model II). After intramuscular injection of procaine penicillin (33,000 I.U./Kg) plasma levels reached a peak at about 270 ng/ml and then declined with a half-life of about 9 hours. These data were approximately fitted by Model II assuming a first order rate constant for absorption of procaine of 0.0024 min-1. After intramuscular injection of procaine HC1 (10 mg/Kg) plasma levels of procaine peaked rapidly at about 600 ng/ml but thereafter declined slowly (+ 1/2 = 2 hours). A satisfactory pharmaco-kinetic model for this intramuscular data could not be developed. An approximation of these data was obtained by assuming the existence of two intramuscular drug compartments, one containing readily absorbable drug and the other poorly absorbable drug (Model III). After intra-articular administration of procaine (0.33 mg/Kg) plasma levels of this drug reached a peak at about 17 ng/ml and then declined with a half-life of about 2 hours. These data were not modelled.


The excretion of ibuprofen by the horse - a preliminary report. The anti-inflammatory drug Ibuprofen [(+/-)-2-(p-isobutylphenyl) propionic acid] was estimated in the blood and urine of a horse using gas-liquid chromatography of the silylated derivative. Levels of the drug in the two body fluids were measured over a period of about 24 hours after administering a 12 gm dose of Ibuprofen. Plasma peak levels were observed within 30 to 60 min, and the drug was no longer detectable in the plasma by 8 hr. Urinary peak levels were observed 200 to 300 min after dosing, and the drug was no longer detectable in the urine by about 28 hr. It was observed that only 2% to 6% of the free unchanged drug was excreted in the urine.


The gas-liquid chromatograph and the electron capture detection in equine drug testing. Three gas-liquid chromatographic (G.L.C.) procedures discussed have been designed around the four "esses" of detection tests--speed, sensitivity, simplicity, and specificity. These techniques are admirably applicable to the very low plasma drug levels encountered in blood testing under pre-race conditions. The methods are equally applicable to post-race testing procedures, where both blood and urine samples are tested. Drugs can only rarely be detected by the electron capture detector (E.C.D.) without a prior derivatization step, which conveys to the drug(s) high electron affinity. Because of broad applicability, two derivatizing agents, heptafluorobutyric (HFBA) and pentafluorpropionic (PFPA) anhydrides are employed. The three techniques, allowing broad coverage of various drug classes are: 1) direct derivatization of drugs to form strongly electron capturing amides and esters. 2) reductive fragmentation of drugs with lithium aluminum hydride to form alcohols, with conversion to ester derivatives. 3) oxidative fragmentation of drugs with potassium dichromate to form derivatizable groups, followed by direct derivatization.


Report on use of XAD resins in racing chemistry. This report comprises a summary of the work done with XAD resin extraction by racing chemists and reported in the Association of Official Racing Chemists publications. It is apparent that the use of XAD resins is becoming more popular in racing laboratories as a technique for routine screening and also for the extraction of certain conjugated drugs. Most laboratories employ variations on the original Brinkmann Drug-Skreen Technique. Comparisons of the efficiency of extraction of drugs from horse urine by XAD-2 resin and by chloroform column extraction indicate that some drugs can be extracted with equal or greater efficiency by the resin technique.


Research and identification of tranquillizers - use of retention index. At the request of the Service des Haras, our laboratory works on the toxicological problems of the sport-horse. These studies have resulted in the setting up of an anti-doping control for equestrian competitions of various types, not only flat racing. During events, horses, must be calm and docile to the riders' order. Frequently, the latter use tranquillizers to try and win events. The analytical method for the research and identification of these compounds is described. The technique involves successively: 1. alkalinisation of the sample - saliva, blood or urine after enzymatic hydrolysis. 2. extraction with diethyl ether - the recovery is 70% to 90% depending upon the drug. 3. determination by gas-liquid chromatography with use of a retention index for qualitative analysis. We can detect up to fifteen tranquillizers in any one sample, even when present at such low concentrations as found in saliva. The use of the retention index is a reliable method for qualitative analysis. For example, the method has been used for three years, during which period the rentention index of acetylpromazine remained at 3240 +/- 7. The chromatographic analysis was performed on 3% OV-17 at 290 degrees. The chromatographic analysis has been performed by three columns of different polarity (OV-1; OV-17; SP-2250). If on the three columns, the retention index of one peak is the same as that of the tranquilizer, a further confirmation is made with the use of a thermionic detector specific for nitrogenous drugs. In conclusion, this method which is sufficiently precise and specific has been used for anti-doping control.


Racing problems in the U.S.A.

Racing problems in the U.S.A. The major problems of racing in the United States at the present time are caused by too much racing. This has led to too few horses and small fields. Consequently many owners and trainers are trying to enter their horses too frequently and to race them when they are not really fit to run. The desire to race horses as frequently as possible has led to constant pressure from horsemen through their organizations for so called "permissive medication". Started in the state of Colorado approximately ten years ago this has grown until finally there are only a few states, notably New York and New Jersey that have resisted the pressure. The drug that gave the opening wedge to permissive medication was phenylbutazone, but this in many states has led to the inclusion of other drugs including analgesics and drugs that veterinarians claim are needed for therapeutic purposes. Some states have endeavoured to control phenylbutazone medication by quantitation and while lower limits cause little difficulty, maximum allowable limits have caused problems and are not practical. While there has been no publicity to my knowledge about frusemide (furosemide, lasix) the abuse of this drug for so called "bleeders" is an example that may seriously interfere with drug detection in urine and its use should be confined to proven "bleeders" (i.e. horses suffering from epistaxis). Pre-race blood testing began roughly ten years ago at the harness tracks and has been resisted by our flat tracks rather successfully up to the present time. The blood testing methods and those used by the same laboratories in post-race urine testing is inadequate and will not detect many illegal drugs.


Doping control in Japan. An automated extraction procedure for the doping test. Horse racing in Japan consists of two systems, the National (10 racecourses) and the Regional public racing (32 racecourses) having about 2,500 racing meetings in total per year. Urine or saliva samples for dope testing are collected by the officials from thw winner, second and third, and transported to the laboratory in a frozen state. In 1975, 76, 117 samples were analyzed by this laboratory. The laboratory provides the following four methods of analysis, which are variously combined by request. (1) Method for detection of drugs extracted by chloroform from alkalinized sample. (2) Methods for detection of camphor and its derivatives. (3) Method for detection of barbiturates. (4) Method for detection of ethanol. These methods consist of screening, mainly by thin layer chromatography and confirmatory tests using ultra violet spectrophotometry, gas chromatography and mass spectrometry combined with gas chromatography. In the screening test of doping drugs, alkalinized samples are extracted with chloroform. In order to automate the extraction procedure, the authors contrived a new automatic extractor. They also devised a means of pH adjustment of horse urine by using buffer solution and an efficient mechanism of evaporation of organic solvent. Analytical data obtained by the automatic extractor are presented in this paper. In 1972, we started research work to automate the extraction procedure in method (1) above, and the Automatic Extractor has been in use in routine work since last July. One hundred and twnety samples per hour are extracted automatically by three automatic extractors. The analytical data using this apparatus is presented below.


The antidoping control in horseraces in Italy. The results and the improvement of the analytical procedures adopted for the control of doping in horses will be reported. This control has been systematically carried out in Italy for about 10 years in the laboratories of Italian Federation of Sport and Medicine in which the biological samples for the control of doping in various sport activities (football, cycling, athletics etc.) are also examined. In this way it is possible to use the same instruments for all these similar problems and compare the results. The analytical procedure is based on the following steps: 1) Extraction of the samples (mainly urine but sometimes blood or saliva). 2) Screening tests by thin-layer chromatography. 3) Confirmatory tests by gas chromatography on different columns and also by gas chromatography coupled with mass spectrometry. These single steps will be separately discussed, and practical problems encountered will be presented.


Less common "doping" agents and substances encountered during routine screening for drugs. The chromatographic and spectroscopic properties of several unusual substances which have been detected in the "alkaloidal" chloroform extract from racehorse urine and saliva samples are reported. Some of these substances have been identified by combined gas chromatography-mass spectrometry and the source of the substance is stated where this is known. Other substances whose identity is not known have been detected and their mass spectra show characteristic amine fragments. The occurrence of these unidentified substances is more frequent in aged urine samples and it would therefore appear that they are associated with putrefaction.


Computer tomography of traumatic extracerebral lesions. Twenty-one traumatic intracranial extracerebral lesions in 15 patients were examined with the EMI scanner using the 160x160 matrix. The results were compared with operation and angiographic findings. Although angiography seems to be a more reliable diagnostic aid, computer tomography has important advantages as it provides information on the nature and extent of the extracerebral lesion as well as on concomitant brain damage.


Myoepithelial activity and the appearances of sialograms: a possible role in the formation of sialectasis. In experimental studies on dogs it has been shown that the sialographic filling of the ductal system may be severely restricted if the myoepithelial cells are contracting and it seems likely that myoepithelial activity in man will also influence sialographic appearances. It has also been found that when secretory forces were accompanied by myoepithelial contraction against a sialographic pressure the ersuing damage to striated ducts produced appearances of sialectasis. IT IS WONDERED WHETHER SIMILAR EVENTS MAY SOMETIMES BE RESPONSIBLE FOR SIALECTATIC APPEARANCES IN MAN.


The röntgenographic findings in the acute neuronopathic form of Niemann-Pick disease. The röntgenographic changes are described in two patients with the acute neuronopathic form of Niemann-Pick disease. These consist of metaphyseal splaying, osteoporosis and the quadrate appearance of the lumbar vertebrae with relatively long pedicles. The parenchymatous involvement is manifested by interstitial lung changes, enlargement of liver, spleen and kidney and distended intestinal loops with an abnormal mucosal pattern associated with prolonged transit time of the contrast material. The differential diagnosis of the above changes is discussed.


Predictive factors and mechanism of arrhythmias and myocardial ischaemic changes in elderly patients during barium enema. The incidence of significant arrhythmias and ST segment changes during barium enema examination (BE) was evaluated by Holter monitoring of 58 unselected patients over the age of 60 years. Forty percent of the group developed new significant arrhythmias of which the most common were frequent and/or multifocal premature ventricular contractions. Seven percent demonstrated ST segment depression. Administration of glucagon did not diminish the incidence of arrhythmias. Predictive factors for ECG abnormalities were advancing age, abnormal pre-BE-ECG, and pre-BE orthostatic hypotension. Analysis of arhythmias and response to phywiologic tension. Analysis of arrhythmias and response to physiologic manoeuvres suggested that the abnormalities were related to increased sympathetic tone which may be intensified by hypovolaemia resulting from routine bowel preparation.


Quantitative aspects of pertechnetate concentration in human parotid and submandibular salivary glands.. Quantitative uptakes of 99Tcm-pertechnetate by the salivary glands have been measured in 36 subjects and normal values established. While uptake over the parotid gland increased with time, parotid uptakes were higher at all time intervals than uptakes over the submandibular glands. The uptake over the submandibulars also increased with time. There was no significant difference between resting uptakes of right and left glands, a significant difference would therefore suggest the presence of pathology. Values over the parotid gland are greater under resting conditions than following stimulation, but no such difference was noted for submandibular glands. However, with stimulated glands it was also shown that there was no significant difference between the right and left parotid, or rignt and left submandibular uptakes. The clinical relevance of these findings is discussed.


Vascular reconstruction in Buerger's disease. In 23 of 148 patients with Buerger's disease, it was possible to undertake 27 arterial reconstructive procedures: bypass in 22 and thrombo-endarterectomy in 5. In a follow-up of 10 months to 8 years, the overall patency rate was 26 per cent. The long term patency rate of bypass grafting was good in obstruction of main vessels, but unsatisfactory with multiple occlusions. Bypass grafting was preferred to thromboendarterectomy. To obtain long term patency of revascularaized segments, complete abstinence from tobacco is absolutely essential. The preparatory manoeuvres for antogenous venous graft should be as atraumatic as possible. A functional diagnosis is indispensable when considering operative indications and for follow-up study of patients with peripheral arterial occlusive disease.


Surgical treatment of kinked internal carotid arteries. Carotid artery insufficiency is caused by an abnormal kinked or coiled internal carotid artery in 15-20 per cent of symptomatic patients. Surgical correction should be considered if other causes for the neurological signs are excluded, a pronounced kinked or coiled area is demonstrated and if there is not a severe neurological deficit. As well as eliminating the elongation, surgical correction should include intraluminal inspection of the artery as, in 38-5 per cent of cases, a concomitant arteriosclerotic stenosis requires simultaneous correction.


Structural deterioration in Celestin tubes. Two cases are described where, after 2 years, a nylon-latex Celestin tube which was left in situ to bypass a benign stricture underwent structural deterioration and produced complications requiring tube replacement. It is recommended that an alternative endo-oesophageal tube should be considered where a lifespan of this order is contemplated.


A practical reappraisal of the anatomy of the extrahepatic bile ducts and arteries. Two hundred and five dissections of the extrahepatic biliary tree have revealed that although the anatomy of this area is remarkably variable there are obly three important vascular and five important ductal anomalies (the former occurring much more frequently than the latter). Each anomaly is, however, easily recognized at operation. A higher incidence of anatomical anomalites was found in patients with gallstones compared with those without and in women than in men, and there was a remarkably low incidence of accessory hepatic ducts overall. The significance and practical implications of these findings are discussed.


Gallstone pancreatitis with normal biliary radiology. Three patients with relapsing gallstone pancreatitis and normal routine biliary radiology are reported and discussed. It is emphasized that when dealing with recurrent pancreatitis for which no cause is evident, normal conventional biliary radiology (oral cholecystogram and intravenous cholangiogram) should not necessarily be accepted as conclusive. The use of endoscopic cholangiography in such a situation is encouraging.


The immunogenicity of starch glove powder and talc. An antigen responsible for cutaneous delayed hypersensitivity to starch glove powder in guinea-pigs was found to be associated with starch grains. Talc was not immunogenic in this system, and did not cross-react in skin tests with starch glove powder. Additives and protein components in the glove powder did not appear to be antigenic.


Allergic starch peritonitis in the guinea-pig. An experimental starch peritonitis in guinea-pigs previously immunized with starch glove powder and Freund's complete adjuvant is described. After peritoneal challenge by 10 mg starch glove powder, 22 per cent of the animals developed florid omental granulomas resembling those found in some patients with post-operative glove powder peritonitis. It is suggested that delayed hypersensitivity to a component of the glove powder may be responsible for the clinical features of starch peritonitis in man.


A comparison of polyglycolic acid and catgut sutures in rat colonic anastomoses. Standard colonic wounds were closed in 44 rats by either polyglycolic acid or catgut sutures. Seven days later portions of the colonic wounds were measured for tensile strength and examined histologically. There was no difference in the mortality, strength of the anastomosis or the amount of inflammation induced by either of the suture materials used.


Tension pneumoperitoneum: a report of 4 cases. Four cases of tension pneumoperitoneum are described. In 3 patients this condition followed a perforation of a grossly distended caecum. In 2 of these patients there was an associated malignant neoplasm of the pelvic colon with obstruction. The third patient had a pseudo-obstruction of the transverse colon. The fourth patient had a tension penumoperitoneum with associated surgical emphysema in the neck and subcutaneous tissues of the abdomen and chest walls, following perforation of a duodenal ulcer. The aetiology, presentation and management, together with the mechanism of tension pneumoperitoneum, are discussed.


Fibrinolytic activator in the endothelium of the veins of the lower limb. The local fibrinolytic acitivity of the femoral, popliteal and soleal veins has been studied using a histochemical technique. The results suggest that the fibrinolytic activity in the soleal veins may be low when compared with that in the femoral and popliteal veins. This may be an aetiological factor responsible for the high incidence of thrombi occurring in the soleal veins.


Bilharzial granuloma of the gastro-intestinal tract. Thirty-four patients with bilharzial granuloma of the gastro-intestinal tract were analysed. The commonest site was the rectosigmoid, Bilharzial "appendicitis" was also common but the exact relation between bilharzial granuloma and acute inflammation remains unclear. Preoperative diagnosis is bilharzial granuloma is occasionally difficult and histological proof is necessary. Surgical treatment with polypectomy in the case of a solitary polyp and excision of the affected segment of bowel when there are multiple polps are recommended. There is no evidence that the condition is premalignant.


Bilhariziasis and prolapse of the rectum. Twenty-six cases with complete prolapse of the rectum were investigated. In 8 of these, bilharziasis of the rectum and colon was found to be the cause of the prolapse. The role of bilharziasis in the pathogenesis of this condition is discussed. The age and sex incidence of bilharzial prolapse differs from that of cases without bilharziasis.


Effect of intravenous dextran 70 and pneumatic leg compression on incidence of postoperative pulmonary embolism. The incidence of pulmonary embolism and deep vein thrombosis was measured in 50 matched pairs of patients undergoing common surgical procedures with preoperative and postoperative ventilation-perfusion lung scans and the fibrinogen uptake test. One patient in each pair was treated with intravenous dextran 70 and pneumatic leggings. The incidence of pulmonary embolism among the treated patients was significantly reduced from 24% to 8%, but the incidence of deep vein thrombosis was not significantly reduced (34% to 24%).


Tobramycin, amikacin, sissomicin, and gentamicin resistant Gram-negative rods. Sensitivities to gentamicin, sissomicin, tobramycin, and amikacin were compared in 196 gentamicin-resistant Gram-negative rods and in 212 similar organisms sensitive to gentamicin, mainly isolated from clinical specimens. Amikacin was the aminoglycoside most active against gentamicin-resistant organisms, Pseudomonas aeruginosa, klebsiella spp, Escherichia coli, Proteus spp, Providencia spp, and Citrobacter spp being particularly susceptible. Most of the gentamicin-resistant organisms were isolated from the urine of patients undergoing surgery. Gentamicin was the most active antibiotic against gentamicin-sensitive E coli, Proteus mirabilis, and Serratia spp. Pseudomonas aeruginosa and other Pseudomonas spp were most susceptible to tobramycin.


Pyorrhoea as cause of pyrexia.

Pyorrhoea as cause of pyrexia. Three patients with fever and malaise, one of whom also had joint pains, were extensively investigated before their condition was attributed to dental sepsis. Each patient recovered fully after appropriate dental treatment. Dental sepsis should be added to the list of possible causes of pyrexia of undetermined origin, and a routine dental examination should be carried out in each case.


Nurses and nursing in primary medical care in England. In 1974 we sent questionnaires on attachment and employment of nurses to 9214 general practices in England. There were 7863 replies (85%), of which 551 were excluded from the study. A total of 2654 nurses were directly employed by 24% (1774) of the practices, and 68% (4972) had attached nurses. Practices in health centres were larger and had greater nursing resources than those in other premises. We suggest that practices may employ nurses to compensate for ineffective nursing attachments, and we conclude that general-practice-employed nurses are becoming "professionalised".


Rational sequence of tests for pancreatic function. Of 144 patients with suspected pancreatic disease in whom a 75Se-selenomethionine scan was performed, endoscopic retrograde pancreatography (ERP) was successful in 108 (75%). The final diagnosis is known in 100 patients and has been compared with scan and ERP findings. A normal scan reliably indicated a normal pancreas, but the scan was falsely abnormal in 30%. ERP distinguished between carcinoma and chronic pancreatitis in 84% of cases but was falsely normal in five patients with pancreatic disease. In extrahepatic biliary disease both tests tended to give falsely abnormal results. A sequence of tests to provide a rapid and reliable assessment of pancreatic function should be a radio-isotope scan, followed by ERP if the results of the scan are abnormal, and a Lundh test if the scan is abnormal but the findings on ERP are normal.


Non-fatal injuries sustained by seatbelt wearers: a comparative study. The injuries sustained by 969 drivers and front-seat passengers in road-traffic accidents were studied. Altogether 196 (20-2%) of the drivers and passengers were wearing seat belts and 773 (79-8%) were not. The injuries among the two groups differed greatly in both severity and distribution. A total of 54 (27-6%) of the seatbelt wearers sustained one or more fractures compared with 300 (38-8%) of the non-wearers, and 18 (9-2%) of the seatbelt wearers were severely injured compared with 300 (38-8%) of the non-wearers. Soft-tissue injuries to the face were sustained by only 29 (14-8%) of the seatbelt wearers compared with 425 (55%) of the non-wearers. Since wearing seatbelts may become compulsory, the type and pattern of injuries to be expected in wearers should be appreciated.


Intensive investigation in management of Hodgkin's disease. Ninety-eight patients with clinically localised Hodgkin's disease underwent laparotomy and splenectomy to determine the extent of microscopic spread. In 68 patients the procedure was carried out for untreated disease apparently confined above the diaphragm. Abdominal disease cannot be confidently excluded on the basis of non-invasive investigation at presentation. Clinical assessment of splenic disease was unreliable unless gross splenomegaly was present. Pedal lymphography was accurate in assessing para-aortic and iliac disease but of no value in assessing other intra-abdominal lymph node involvement, including that of the mesenteric lymph node. Trephine bone marrow biopsy findings were normal in all patients before surgery, and only one patient was found to have diseased bone marrow by Stryker-saw biopsy at operation. Liver disease was identified at operation in nine patients, some of whom were asymptomatic with clinically undetectable splenic and nodal disease. Detailed clinical staging failed to detect disease in one-third of patients who underwent laparotomy. These studies show that if radiotherapy is to remain the treatment of choice for disease truly localised to lymph nodes a detailed staging procedure, including laparotomy and splenectomy, remains essential. The value of this potentially curative treatment is considerably diminished in the patient who has been inadequately staged.


Increased insensible water loss in newborn infants nursed under radiant heaters. Urine osmolality was studied in 38 babies nursed in conventional incubators or cots and 18 nursed under an overhead radiant heat shield. Among 50 babies receiving a similar fluid intake in the first 48 hours of life mean urinary osmolality was significantly higher in the radiant heater group. In babies weighing less than 1500 g a trend towards higher urinary osmolalities was recorded in those nursed under radiant heaters even though they had received amost double the fluid intake of the incubator group. Severe hypernatraemia occurred in four of the five babies weighing less than 1000 g who were nursed under radiant heaters but in none of the seven babies of similar birth weight nursed in incubators. These findings are consistent with previous observations of an increase in insensible water loss in babies nursed under radiant heaters and emphasise the importance of providing enough extra water for these infants and the need for close monitoring of their fluid balance. The latter may be done at the bedside by measuring urinary specific gravity with a hand refractometer.


Vitamin D status in different subgroups of British Asians. To assess the effect of religious dietary practices and social customs on the vitamin D status of Asian immigrants, we kept records of the dietary intake and time spent out of doors of 81 Ugandan Asian men, women, and girls (9-19 years old). Sera were analysed for 25-hydroxycholecalciferol (25-OHD3), and 28% of the subjects were found to have levels below the lower limit of normal. The (vegetarian) Hindus had the lowest dietary intakes, least time out of doors, and lowest serum 25-OHD3. The Goan (Roman Catholic) Asians, despite more pigmentation, had 25-OHD3 levels similar to those found among indigenous British people and had the most satisfactory vitamin D intakes. Among Asians, whose exposure to sunlight may be limited, dietary vitamin D becomes the major determinant of serum 25-OHD3.


Changed levels of endogenous sex steroids in women on oral contraceptives. Serum and urinary levels of unconjugated testosterone, dihydrotestosterone, and oestradiol were measured by specific radioimmunoassays in 10 healthy women in the early follicular phase of their menstrual cycle and in nine healthy women taking oral contraceptives. The contraceptive group had testosterone levels 1-3 times higher and dihydrotestosterone levels two times higher than those in the controls. Serum oestradiol levels in the contraceptive group were much lower than those in the controls and similar to levels in postmenopausal women. The contraceptive group had about twice the urinary excretion of unconjugated (free) testosterone and dihydrotestosterone of the controls, but their excretion of unconjugated oestradiol was 2-7 times lower. The great increase in serum and urinary androgen concentrations, as well as the suppression of oestradiol, may be related to the antiovulatory effect of oral contraceptives.


Weak autoantibody reactions to antigens other than sperm after vasectomy. Autoantibody activity against various antigens was measured by indirect immunofluorescence in 97 men about to undergo vasectomy and 170 men who had undergone the operation up to six years earlier. There was a significantly higher prevalence of weakly positive autoantibody reactions among those who had undergone vasectomy. There was, however, no evidence that vasectomy could induce stronger autoantibody reactions such as those associated with autoimmune disease.


Computerised tomography of acute traumatic intracranial haematoma: reliability of neurosurgeons' interpretations. Two neurosurgeons concerned with the emergency management of patients with head injury correctly diagnosed the presence or absence of an acute intracranial haematoma in 97 scans that were presented to them without knowledge of the patients' clinical details. There were no false-positives or false-negatives, although identification of the type of haematoma was not always possible. The impact of the EMI scan on patient management demands new approaches to the care of head injuries.


Prednisone in MOPP chemotherapy for Hodgkin's disease. High remission rates have been produced by MOPP (mustine, vincristine, procarbazine, and prednisone) chemotherapy in patients with advanced Hodgkin's disease, but the prednisone component has caused adverse effects in patients who have undergone radiotherapy. The remission rates and length of remission were reviewed in 211 patients with Hodgkin's disease who received chemotherapy either with or without prednisone. In contrast to the findings of a British study, there were no significant differences in remission rates or length of remission between patients who had received prednisone and patients who had not. There were differences between the British prospective study and this retrospective one, but it is difficult to know what accounted for the substantial differences in the findings.


Abnormal calcium metabolism in normocalcaemic sarcoidosis. In studies of calcium metabolism in 13 unselected patients with untreated sarcoidosis all were normocalcaemic but five had hypercalcuria. All had normal renal function. Calcium absorption was indexed by a double isotope test. 45Ca hyperabsorption occurred in six patients. Ten kinetic studies were carried out with 47Ca and in six bone turnover was increased. 45Ca absorption correlated well with the calculated bone uptake rate of calcium, and with urine calcium excretion. These results suggest that in sarcoidosis abnormalities in calcium metabolism are fairly common although they rarely result in sustained hypercalcaemia.


Coagulation and fibrinolysis in intact hydatidiform molar pregnancy. Tests of coagulation, fibrinolysis, and platelet function were performed in 17 patients with intact molar pregnancies. Women with intact molar pregnancies had higher fibrinogen factor VIII, and fibrinogen degradation products, concentrations and lower prothrombin, factor X, plasminogen, and plasminogen activator concentrations than controls with normal pregnancies. They also had reduced platelet counts and thromboelastographic values, which indicated hypocoagulability. These results suggest that intravascular coagulation occurs in intact hydatidiform molar pregnancies.


Regional survey of femoral neck fractures. In the South-west Thames Region 2619 patients (2105 women and 514 men) were discharged with a diagnosis of femoral neck fracture in 1974. The equivalent of a 250-bedded hospital was occupied throughout the year. The incidence, average length of stay, and mortality rate rose with increasing age and there were differences in these indices in the five health areas. These results confirm the enormous burden placed on the hospital service by patients with fracture of the femoral neck but suggest that differences in practice in the five areas may contribute to the size of the problem.


Organization of auditory cortical areas in man. Average responses to clicks were recorded from the exposed human cortex of 19 adult patients during operation for the treatment of intracranial diseases. Auditory evoked responses were obtained from two areas. Short latency potentials were recorded on the superior surface of the temporal lobe corresponding to the transverse temporal gyri. These responses consisted of two positive waves P1 and P2 separated by a negative wave N1. P1 had a mean latency of 14-7+/-1-5 ms, N1 a mean latency of 19-1+/-2-6 ms and P2 a mean latency of 32-2+/-4-1 ms. Responses to stimulation of the contralateral ear were of higher amplitude than responses to stimulation of the ipsilateral ear. Responses of smaller amplitude and longer latency were obtained from the superior temporal gyrus and the upper lip of the sylvian fissure. These responses had a mean peak latency for P1 of 40-2+/-2-6 ms, for N1 of 62-5+/-12-5 ms and for P2 of 97-7+/-17-2 ms. It is concluded that the cortical auditory region of man may be subdivided in two major areas: an area on the supratemporal plane representing the primary auditory area or A1 and a region surrounding A1 which perhaps comprises two areas, one on the superior temporal gyrus and one on the upper bank of the sylvian fissure including frontal and parietal operculi.


A case of mixed transcortical aphasia with intact naming. Altholgh Lichtheim recognized that Wernicke's 'reflex arch' (primary auditory area, to Wernicke's area, to Broca's area, to primary motor area) was important for repetition, he recognized that other areas of the brain (for example, area of concepts or semantic area) must be important in comprehension and voluntary speech. He suggested that Wernicke's area (phonemic area) not only projected to Broca's area (as Wernicke suggested) but that it also projected to the area of concepts. A lesion of this latter pathway or in the area of concepts would produce a syndrome where repetition was intact but comprehension was impaired (e.g. transcortical sensory aphasia). Lichtheim also thought that the area of concepts projected directly to Broca's area and that voluntary speech was mediated by this pathway. Although Lichtheim's model could explain the mechanism underlying transcortical aphasia, his schema could not explain anomic aphasia. Unlike Lichtheim's schema, Kussmaul's schema suggested that the area of concepts projects back to Wernicke's area before projecting to Broca's area. With this schema, a patient with a hypothetical lesion which interrupted the pathway from the area of concepts to Wernicke's area (but did not interrupt the pathway from Wernicke's area to the area of concepts) should be anomic, with normal comprehension and repetition. In order for this latter schema to be plausible there should also be a lesion which interrupts the pathway from Wernicke's area to the area of concepts but does not interrupt the pathway which goes from the area of concepts to Wernicke's area. A patient with this hypothetical lesion should comprehend poorly; however, in spite of poor comprehension, naming and repetition should be intact. We report a patient who demonstrates poor comprehension with intact naming and repetition. This patient could also read aloud but could not comprehend written language. Not only could this patient name objects but he could demonstrate their use. These observations suggest that comprehension of written language is mediated by a different pathway than the recognition of visually presented objects.


The involvement of noradrenaline in motor activity as shown by rotational behaviour after unilateral lesions of the locus coeruleus. Unilateral electrolytic lesions in the region of the locus coeruleus in rats result in contraversive circling behaviour following the administration of systemic apomorphine or dexamphetamine. In a large group of such animals there was a very good correlation between histological damage to the locus coeruleus and the presence of such rotation. There was a progressive increase in the percentage of rats showing this drug-induced behaviour as ipsilateral cerebral cortical noradrenaline content decreased. Unilateral injections of 6-OHDA produce more selective damage to the noradrenergic locus coeruleus and result in a similar, although less marked, apomorphine- and dexamphetamine-induced rotation. Unlike the rats with electrolytic lesions of the locus coeruleus, those with 6-OHDA injections did not show an increase in ipsilateral striatal dopamine content. It seems likely that the motor asymmetry seen with lesions which involve the locus coeruleus is due to damage to its noradrenergic cells. A possible explanation of the differences between electrolytic and 6-OHDA induced lesion is that the 6-OHDA produces damage to the ventral noradrenergic bundle as well as the locus coeruleus and results in an increase in ipsilateral cerebral cortical 5-hydroxytryptamine.


Automatisms during frontal lobe epileptic seizures. Three new cases of automatisms occurring during frontal lobe epileptic seizures are reported. If these cases are added to those already published and adequately described in the literature, a total of 12 cases is obtained. An attempt is made to elucidate the clinical characteristics of automatisms encountered during frontal lobe seizures. This kind of automatism would seem to be clinically distinguishable from other types.


Disorders in cerebellar ocular motor control. I. Saccadic overshoot dysmetria. An oculographic, control system and clinico-anatomical analysis. Photoelectric eye movement recording in 9 patients with cerebellar disorders defined three features of saccadic overshoot dysmetria: (i) saccades were hypermetric and successively diminished in amplitude; (ii) saccadic initiation interval averaged 173 ms: and (iii) eye position was constant during the intersaccadic period. These characteristics indicated that the visually evoked saccades subserving foveation had increased gain, and were modelled by computer simulations using a sampled-data control model with increased feed-forward gain. Eight patients with saccadic overshoot dysmetria had cerebellar neoplasms, vermis-splitting surgical procedures and mid-line cerebellar signs. This clinical evidence suggests that vermian dysfunction is responsible for saccadic overshoot dysmetria. Normally, the cerebellar vermis appears to play an adaptive role by continuously adjusting gain of the direct visual motor pathway. When cerebellar disorder exists, adaptive gain modulation is lost, and, if gain then increases, saccadic overshoot dysmetria is a result.


Disorders in cerebellar ocular motor control. II. Macrosaccadic oscillation. An oculographic, control system and clinico-anatomical analysis. A distinctive cerebellar ocular motor disorder, macrosaccadic oscillation, evolved simultaneously with an acute cerebellar syndrome in 4 patients, 2 with haemorrhagic metastatic melanoma deep in the vermis, a third with a presumed cerebellar haematoma and a fourth with focal demyelinating disease. Ocular oscillations were conjugate, horizontal, symmetrical, occurred in bursts of several seconds duration, had amplitudes of 30 degrees to 50 degrees, and were evoked whenever the patient attempted to shift visual fixation or pursue a moving target. Photo-electric recordings in one patient with tumour defined features of this disorder of saccadic eye movement: (i) oscillation was composed of saccades, (ii) frequency was 2 Hz, (iii) bursts occurred with amplitude first increasing and then decreasing, (iv) intervals between beginnings of saccades averaged 260 ms and (v) eye position did not exhibit systematic drift during the intersaccadic period. These features documented the inreased gain and instability of the visually guided saccadic system. By using increased feed-forward gain in a sampled-data control model we simulated the pattern of macrosaccadic oscillation. We belive that the acute loss of the calibrator function of the cerebellum accounts for the gain abnormality underlying macrosaccadic oscillation.


Cerebral blood flow and metabolic rate early and late in prolonged epileptic seizures induced in rats by bicuculline. Cerebral blood flow (CBF) and cerebral metablic rate for oxygen (CMRO2) have been studied during sustained epileptic seizures induced by bicuculline (1-2 mg/kg, i.v.) in paralysed Wistar rats, artificially ventilated with nitrous oxide/oxygen. CBF was determined by venous outflow collection, and by 133Xe desaturation, using sagittal sinus blood (for cerebral cortical flow) or retroglenoid venous blood (for 'whole brain' flow). The procedure employed ensured that arterial oxygenation remained normal and blood glucose concentration was normal or high throughout the seizure. Arterial hypotension was prevented by the infusion of donor blood. CBF increased concurrently with seizure onset, reaching a maximum nine times higher than control value after 15-60 s. This was due to a marked rise in mean arterial pressure (to greater than 180 torr) and a dramatic fall in cerebrovascular resistance to less than 15 per cent of control). Subsequently, with decreasing blood pressure, CBF slowly diminished, being more than four times higher than control at 20 min, and slightly less than three times higher than control at two hours. The different procedures for measuring CBF gave closely similar results. A threefold increase relative to control CMRO2 (7-6 ml/100 g-1/min-1 for 'whole brain,' and 10-2 ml/100 g-1/min-1 for cerebral cortex) was measured after 1-20 min of seizure activity (utilizing either the venous outflow or the 133Xe desaturation procedure for CBF determination). After two hours of seizure activity CMR02 was still more than twice as high as the control. This high metabolic rate during sustained seizure activity will increase the susceptibility of the brain to 'ischaemic' damage during prolonged seizures in man in which an additional metabolic stress may be imposed by cerebral hypoxia, arterial hypotension, hyperpyrexia or hypoglycaemia.


Auditory brain-stem responses in brain death. Auditory brain-stem responses were measured by far-field recording techniques in 27 patients fulfilling the criteria of brain death. The responses were either absent or consisted of the presence of just the initial component (Wave I). Wave I, when present, was of normal amplitude but prolonged in latency. Four patients were followed over several days from a state of coma with evidence of preserved brain-stem and cerebral functions to a clinical state compatible with brain death. Auditory brain-stem responses were initially intact and then showed a decrease in amplitude and a prolongation of latency of the later components until finally Wave I was alone. Auditory brain-stem responses are an objective measure of one of the sensory pathways traversing the brain-stem and can be used to evaluate the functional states of the brain-stem in patients in whom the question of brain death has been raised.


Reciprocal Ia inhibition in spastic hemiplegia of man. The results of the present electrophysiological investigation have shed some light on the mechanisms underlying many clinical signs, at least, in patients with capsular hemiplegia. A tentative interpretation of them is given below. Cerebral lesions due to haemorrhage or infarction in the area of the middle cerebral artery interrupt an extensive part of the corticospinal tract and disturb many other descending pathways involved in voluntary performance. In consequence, a marked reduction in the ability to drive the spinal motor apparatus occurs, resulting in weakness of motor power. Here, we refer only to muscle power but not to performance. For example, the disturbance of voluntary contraction by clonus is disregarded (cf. fig. 8). On the other hand, the same lesions also release the spinal reflexes from inhibition by the higher levels of the brain and cause increased excitability in flexors and extensors. In the lower extremity, this is much more makred in extensors and extensor spasticity becomes a dominant sign clinically. Any release effect on the flexor system is largely cancelled by the high activity of the reciprocal Ia inhibitory pathway from extensors and only a fragment of it is occasionally revealed in some patients as an H-reflex in pre-tibial muscles or as weak Ia inhibition of the triceps surae. Reduced driving power of the brain may be compensated by raised excitability in the spinal cord and spastic extensors are thus naturally in a better condition to preserve motor power. Flexor muscles are doubly crippled by reduced descending impulses and strong reciprocal inhibition by the Ia impulses from the spindles of the extensor muscles.


Kinetics of horseradish peroxidase migration through cerebral cortex. This study provides a semiquantitative description of the migration of the extracellular space marker, horseradish peroxidase (HRP), through the cerebral cortex. Following a continuous subarachnoid infusion of HRP, this marker was fixed rapidly within the cortical extracellular space (ECS) by intravascular aldehyde perfusion fixation. Microscopic measurements of the maximum depth of penetration of HRP into the cortex perpendicular to the pial surface were taken from coronal whole brain sections of rabbits that had been exposed to HRP for varying periods of time. The depth of penetration plotted as a function of time of exposure to HRP produced a 'diffusion profile'. The failure of experimental points to conform to an ideal diffusion curve indicates that simple diffusion alone is an inadequate explanation of the rate of movement of large molecular solutes from the subarachnoid space through the cerebral cortical ECS. The complex pattern of migration velocity of HRP through cortex may be due to alterations of flux of this solute which result from variations in the volume of ECS in different cortical laminae and to the presence of bulk flow of extracellular fluid in the deeper cortical regions.


The genetic organization of neuron number in Ammon's horns of house mice. This study of the Ammon's horns of 20 inbred strains of house mice had 3 primary objectives. The first was to determine whether there was genetically-associated variability in the number of neurons present within a dorsal and a ventral portion of this brain region. If genetically-associated variability proved to be present, there were two additional objectives which we wished to achieve. One was to determine the degree to which genetically-associated variations in neuron number are confined to specific neuronal regions, or shared between neuronal regions. The other was to identify specific extreme strains for further study. A section cut in a coronal plane was selected for a sampling within a dorsal portion of Ammon's horn, and a section cut in a horizontal plane was selected for a sampling within a ventral portion. Counts were made of neuronal nuclei in the granule cell layer of the dentate gyrus and in the pyramidal layers of regio inferior and regio superior of the hippocampus. Cross-sectional areas for these somal laminae were also measured and planimetric neuron densities were obtained by computing the ratios of nuclei counted to the areas of the appropriate somal laminae. Genetically-associated variability in counts of neuronal nuclei was observed for all neuronal regions within both the dorsal and the ventral sampled portions. Genetically-associated variability in planimetric neuron density was clearly present within the dorsal portion. The obtained counts of neuronal nuclei were adjusted statistically for variations in planimetric neuron density to provide the best indices of variation in neuron number presently available. Results for these adjusted neuron counts were as follows. First, there was an indication of substantial genetically-associated variability in neuron number. With the single exception of one neuronal region (dorsal regio superior), adjusted neuron counts exhibited significant genetically-associated variability for all neuronal regions within both portions sampled. Second, there was evidence for both separate and shared genetic determination of neuron number between neuronal regions. Strains were not uniformly high, intermediate, or low in their adjusted neuron counts. Instead, a complex patterning of high and low genetically-associated correlations was observed between neuronal regions and sampled portions. Third, extreme strains for each neuronal region were identified for the sampled dorsal and ventral portions. Differences between the neuron numbers of interconnected neuronal regions imply variations in connectivity. Such variations could provide a morphological basis for 'tuning' associated adaptive characteristics of populations of house mice to enhance the likelihood of their survival in environments with varying ecological requirements.


Generators and topography of hippocampal theta (RSA) in the anaesthetized and freely moving rat. Spontaneous and hypothalamically induced hippocampal rhythmical slow activity (RSA or theta) was studied acutely in rats anaesthetized with urethane or immobilized with D-tubocurarine. Systematic tracking of microelectrodes showed two foci of hippocampal RSA, one located in the basal part (stratum oriens) of CAl (mean amplitude 1 mV) and the other located in stratum moleculare of the dorsal blade of the fascia dentata (mean amplitude 2 mV). The hippocampal RSA recorded from the lower blade of the fascia dentata was always smaller than that found in the upper blade (mean amplitude 1 mV). The whole dorsal hippocampal extent within each generator zone was shown to be in synchrony, and the respective generator zones of both hippocampi were synchronous with one another. A null zone in stratum radiation was found interposed between the two generators and a zone of large amplitude fast activity (30-50 Hz) was localized to the hilus of the fascia dentata. Wave form analysis showed that the RSA recorded from the two generators was approximately 180 degrees out of phase. Amplitude and analysis of phase changes of RSA recorded in brain areas outside of the two generator zones suggested that such activity was due to physical spread from the two generators, with the possible exception of a restricted portion of CA3. The existence of the two generators, 180 degrees out of phase, was demonstrated in freely moving rats. Behavioural observations showed that the two generators were related systematically to concurrent motor behaviour. Preliminary observations suggest that, of the two generators, the one located in CAl may be the more variable.


The effect of 5-bromodeoxyuridine on the postnatal development of the rat cerebellum: a biochemical study. The effect of a thymidine analog, 5-bromodeoxyuridine (BrdU), on the postnatal development of the cerebellum was studied. Rats were injected with 15 mg per 100 gram body weight of BrdU twice a day for 3 consecutive days from the second day after birth, and were killed at 5, 7, 15, 22 and 35 days of age. One hour prior to killing, the rats were given tritiated thymidine. The cerebellar DNA, RNA, protein and cerebroside, and the incorporation of thymidine were measured. BrdU administration caused a markedly retarded growth of the body and the cerebellum. At 35 days of age, the weights of the body and the cerebellum were 42 and 69% of the controls. Quantitative measurements of cerebellar nucleic acids and isotope uptake correlated well with previous morphological observation, and indicated that the analog caused a transient inhibition of cell formation and possibly destruction of stem cell population of the external granular layer. This inhibitory effect was compensated later by a more rapid DNA deposition and a prolongation of the period of cell proliferation. However, the restitution was incomplete and resulted in a permanent deficit in the final cell number, as reflected by the reduction in the size of the cerebellum of the BrdU-treated rats.


Clinical repercussion of the various states of extrinsic pulmonary granulomatoses on the bronchial tree. The AA. evaluated the importance of the bronchial participation in 175 patients with histologically confirmed pulmonary granulomatoses. Many had symptoms specifically due to bronchial changes. In some of the patients, bronchial symptoms appeared late in the natural development of the disease, while in others they filled the clinical picture from the beginning. The AA. discuss the possible influence of other aggressors of the bronchial mucosa, e.g. tobacco, and conclude that, in a fair number of cases, it was not possible to attribute the pathology to them. In a patient with a history suggestive of pathology due to inhaled foreign particles, even when bronchial symptoms predominate, an extrinsic pulmonary granulomatosis must be considered for diagnosis.


The role of bronchoscopy in the study of extrinsic pulmonary granulomatoses. As the bronchial mucous membrane is one of the chief barriers against aggression to the respiratory tract by inhaled particles, the AA. reviewed 68 patients with pulmonary granulomatoses studied at the Department of Chest Diseases, Lisbon University Hospital, that had been submitted to bronchoscopy. Only about 22% of these had normal appearing mucosa, while the rest, among other aspects, showed congestion and edema. Bronchial biopsies were taken in the majority of patients (44). Only in two cases was the mucosa considered histologically normal while granulomas were found in 36% and generally it was possible to identify the nature of the foreign particles inhaled. Analysing the results, the AA. conclude that organic particles seem to produce granulomatous reactions more frequently, while inorganic particles seem rather to produce a marked irritating effect. The frequency with which bronchial changes were found in these patients seems to warrant the conclusions that bronchoscopy plays an important role in the study of patients with extrinsic pulmonary granulomatoses.


Effects of dietary dilution with fibre on the food intake and gut dimensions of Japanese quail. Japanese quail (Coturnix coturnix japonica) were fed on either a conventional diet (low-fibre) or the same diet diluted with 200 g oak sawdust/kg (high-fibre). The lengths of the colo-rectum, small intestine, two caeca combined and empty gizzard weight were measured at 10 and 20 weeks of age. Food intake and rate of passage were measured and an approximate measure of digestibility was also made. 2. There were separate effects of dietary fibre, sex and age on gut size. It was larger in birds fed on high-fibre diet, in females and in older birds. 3. Increased dietary fibre caused increased food intake, but had no obvious effect on the rate of passage. 4. When quail were fed on the low-fibre diet or the same diet diluted with 200 or 400 g cellulose powder/kg gut size at 20 weeks of age was greatest when the most fibrous diet was fed and decreased concomitantly with dietary fibre concentration. 5. It appeared that differences in gut size between dietary treatments were more a reflection of variations in food intake than of fibre per se, and that the effects of sex and age on gut size were probably similarly explained.


Changes in food intake and gut size in Japanese quail in response to manipulation of dietary fibre content. Adult Japanese quail (Coturnix coturnix japonica) which had been fed, since hatching, on either a conventional diet (low-fibre) or on the same diet diluted with 200 g oak sawdust/kg (high-fibre) took 8 to 10 d to adjust their food intake when their diets were interchanged. 2. Those fed on high-fibre weighed less than those fed on low-fibre diet, and body weight changed rapidly when the diets were interchanged. All the birds digested about the same weight of nutrients in a day, irrespective of diet, and this suggests that body weight differences were due mainly to differences in energy utilisation, birds fed on high-fibre diet having to expend more energy on feeding than those fed on low-fibre diet. 3. Gut size was greater in birds fed on high-fibre diet, presumably to accommodate the greater bulk of food eaten, and when the diets were interchanged the gut dimension of both groups changed at similar rates, reaching the appropriate sizes for the respective diets in 3 to 4 weeks.


Effect of dietary vitamin A, ascorbic acid and their interaction on turkey bone mineralisation. At early ages, added dietary ascorbic acid did not affect growth or femur mineralisation of poults. In poults after 8 and 12 weeks, femur mass was reduced by 3000 mg ascorbic acid per kg diet. 2. Plasma calcium and magnesium were not influenced differently by 1000, 4000 and 16,000 IU vitamin A/kg diet, but plasma phosphorus was decreased by the 16,000 level. 3. Vitamin A at 1000 or 16,000 IU/kg diet reduced growth in both sexes to 12 weeks of age compared with the NRC recommended 4000 IU/kg diet. In males, the growth retardation continued to 18 weeks. 4. Vitamin A at 16,000 IU/kg reduced femur development as measured by femur weight to body weight or to femur length ratios.


An assessment of the optimal range of dietary sodium for inducing a pause in laying. Laying hens were fed on a diet containing 0-9 g sodium/kg until 34 weeks of age, then groups were fed on diets containing 0-2, 0-3,0-4, 0-5, 0-6 or 0-9 g sodium/kg for 16 weeks before that containing 0-9 g sodium/kg was reintroduced for all birds. 2. Egg production and food consumption were depressed by the low-sodium diets in proportion to the dietary sodium content. 3. Birds receiving 0-3 to 0-6 g sodium/kg diet lost weight initially but subsequently gained weight; birds receiving 0-2 sodium/kg diet lost weight continouosly. 4. In birds receiving 0-2 g sodium/kg diet, the reproductive organs were completely regressed, whereas these organs resembled those of point-of-lay pullets in birds fed on 0-3 or 0-4 g sodium/kg. 5. When the control diet was reintroduced, birds which had received the low-sodium diets resumed normal egg production and food consumption and regained body weight. 6. The optimal range of dietary sodium for inducing a pause in egg laying is 0-3 to 0-4 g/kg.


Aspects of lipid and carbohydrate metabolism, dietary biotin and fatty liver and kidney syndrome in broilers. Two experiments were performed to investigate the biochemical changes associated with FLKS. A diet reported to induce FLKS was fed with or without supplementary biotin to broilers. In experiment 1 various stresses were applied to the birds. 2. In experiment 1 mortality from FLKS was 6% and in experiment 2 nil. Stress had little effect on the induction of the syndrome. 3. There were no significant differences due to diet in any of the variables examined in apparently normal birds. 4. Birds affected by FLKS showed the typical changes of increased liver and kidney weights and lipid contents but hepatic enzyme activities did not differ significantly from those of normal birds except that malate dehydrogenase (decarboxylating) (NADP) activity was significantly decreased. 5. Despite the low content of biotin in the unsupplemented diet (57 mug/kg) liver biotin content was not low in birds fed on this diet. 6. The results suggest that the incidence of FLKS is not related solely to dietary biotin content.


The effect of pregnenolone, progesterone, deoxycorticosterone or corticosterone on the time of ovulation and oviposition in the hen. Pregnenolone, progesterone, deoxycorticosterone and corticosterone were injected into hens between 16.30 and 17.00 h on the day of oviposition of the last egg of a sequence. 2. Pregnenolone did not affect ovulation, but all of the other steroids induced ovulation prematurely. 3. To induce premature ovulation in 50% of the hens, 82-31 mug+/-0-06 mug, 23-86 mug+/-0-07 mug and 659-26 mug+/-0-05 mug of progesterone, deoxycorticosterone and corticosterone, respectively were required for injection. 4. Differences were observed in the times of oviposition of eggs which ovulated in response to injections of either progesterone or a corticosteroid and it was suggested that the mechanism of action of progesterone and corticosteroids operates through different endocrine pathways.


Open lung biopsy as a diagnostic adjuvant in the diagnosis of pulmonary diseases. In a survey of the possible means of establishing a diagnosis in diffuse pulmonary diseases, the usefulness of the open lung biopsy technique is discussed in more detail. In the opinion of the writers the open biopsy offers advantages over the other biopsy methods since: 1) a greater number of and more extensive biopsies can be taken; 2) the hazard of complications is slighter; 3) differences in biopsy specimens, for example, from the upper lobe and the lower lobe can be studied; 4) multiple biopsy specimens permit the carrying out of many examinations such as virological examination, bacteriological examination, histochemical examination, electron microscopic examination and immunofluorescence examination. The method under discussion carries no risk. There was no mortality in this series. The hazard of complications is slight and certainly slighter than in other methods; our experience has shown that it is slighter than in other methods of biopsying the lung.


Temporal and thermal effects on deformation potentials in bone. There were 44 freshly excised adult rat femora subjected to mechanical deformation in a 4-point bending jig while the load/deformation curves and the electrical potentials generated were recorded simultaneously. Serial testing was repeated after storage of specimens up to 30 days at 25 degrees C and -15 degrees, and after heating for 1 h at 70 degrees, 80 degrees, and 85 degrees C. The amplitude of the potential decreased shortly after the femora had been excised until, by 4-7 days, it reached a plateau which was 10-20% of the initial value. In some cases polarity reversal occurred. These changes were not prevented by freezing or drying nor were they accompanied by increased stiffness. It is presumed that they are related to more subtle changes in the mechanical properties of bone or to alterations in the generating or conducting mechanisms. Heating caused a significant increase in potential without a change in the waveform. A corresponding increase in the deformation occurred due to softening of the bone which could, in itself, be responsible for the increased potential without a basic alteration in the transducer mechanism.


The biological activity of synthetic 25,26-dihydroxycholecalciferol and 24,25-dihydroxycholecalciferol in vitamin D-deficient rats. The biological activity of synthetic 24,25 and 25,26 diOHD3 was studied in vitamin D-deficient rats. The purpose of this study was to investigate the influence of small doses of both metabolites (0.125-0.250 mug) upon intestinal calcium transport and bone calcium mobilization. Both metabolites were able to increase calcium absorption in rats maintained on a calcium-deficient diet, but failed to do it in rats on a normal calcium diet. Bilateral nephrectomy suppressed this effect. The "bone calcium mobilization" of both derivatives was measured in vitamin D and calcium- or phosphorus-deprived rats after one intravenous dose. When serum calcium was initially low, 24,25 and 25,26 diOHD3 increased serum calcium moderately, but the increment was only significant with 24,25 diOHD3. When serum calcium was normal before the injection, both metabolites decreased serum calcium significantly, and the decrease was greater with 24,25 diOHD3. Intraperitoneal administration of the metabolites for 5 consecutive days produced a significant increase of calcium in serum and bone ash.


Electron probe microanalysis of secondary carious lesions adjacent to silicate fillings. Secondary caries associated with silicate fillings is characterized by lesions at the tooth surface and lesions of the cavity wall. The mineral content of the cavity wall lesions and the penetration of elements originating from the silicate fillings were studied in experimental in vitro and in vivo lesions as well as in natural carious lesions. Dentine wall lesions, where microradiographs had shown increased radiopacity relative to intact tissue, exhibited increased Ca and P values. Elements derived from the silicate fillings were regularly found in enamel and dentine. The concentrations of fluorine (F) and of zinc (Zn) amounted to 2-3% by weight close to the cavity wall and decreased gradually to less than 0.1% at depths of 600 mum and 400 mum, respectively, from the cavity. Aluminum (Al) most often occurred in a 20-40 mum-wide zone, showing a maximum concentration of 2-3% near the cavity. Sulfur (S) was often present in the dentinal cavity walls of natural secondary caries, but not in the in vitro specimens. The findings indicate that F released from the silicate filling significantly modifies the progress of a carious lesion in the adjacent enamel and dentine. Through its tendency to form complexes with F, Al may possibly enhance the cariostatic effect of F.


Effect of bicarbonate feeding on immobilization osteoporosis in the rat. A time course study of immobilization osteoporosis was carried out in an experimental model in which the hind half of the rat was paralyzed by spinal section between the 3rd and 4th lumbar vertebrae. The earliest changes in the femure were a reduction in water content and in bone volume (3 days after operation). A significant reduction in both hydroxyproline and phosphate content was observed 8 days after immobilization. Calcium, magnesium, sodium, citrate, and carbonate were found significantly decreased at 15 days. Bone potassium remained unchanged. The rate of calcium accretion and the size of the exchangeable pools, when expressed per unit of bone mass, were not significantly affected by treatment. Sham-operated rats fed a bicarbonate-enriched diet, showed increased rates of calcium accretion and bone calcium content. The latter was not accompanied by increases in phosphate and carbonate contents. Operated animals fed the same diet showed an increased rate of bone resorption attributed to an assumed higher rate of parathyroid hormone secretion induced by bicarbonate feeding.


1,25-Dihydroxycholecalciferol increases bone resorption in thyroparathyroidectomised mice. Mice, 1 week old, prelabelled with 45Ca, were either thyroparathyroidectomised or sham-operated; 1 day later half of the mice of each group were injected with 1,25-dihydroxycholecalciferol (5 ng/g), and 20 h later all the mice were killed. Bone resorption in explants was then measured by an in vivo/in vitro technique previously published; compared with untreated mice it was found that 1,25-dihydroxycholecalciferol had increased resorption irrespective of whether the mice had been thyroparathyroidectomised or not. These data suggest that 1,25-dihydroxycholecalciferol is able to increase bone resorption independently of parathyroid hormone.


A morphometric investigation of the duodenal mucosa of normal, vitamin D-deficient, and vitamin D-replete rats. Light and electron microscopy were utilized to determine differences in morphology and size in duodenal villi and microvilli from normal, vitamin D-deficient, and vitamin D-replete rats. An increase in villus diameter and surface area was observed in vitamin D-deficient rats. Administration of vitamin D reversed the increase in diameter but the height of villi increased resulting in longer, thinner villi without a significant change in surface area. A decrease in microvillus surface area seen in vitamin D-deficient animals was reversed by the administration of vitamin D.


Presence of intracisternal type A particles in normal bone tissue from various mouse strains. Intracisternal type A particles were detected by electron microscopy in cells of normal trabecular bone tissue from the mouse strains C57BL, NMRI, C3H, 101, and (C3H X 101)F1. The relatively small numbers of particles observed in all five strains were roughly equivalent. About 10% of the bone cell profiles examined in specimens from each animal contained a single particle. The intracisternal type A particles occurred predominantly in osteoblasts and rarely in osteoprogenitor cells and osteocytes. They were not found in osteoclasts.


Plasma proteins present in human cortical bone: enrichment of the alpha2HS-glycoprotein. The spectrum of plasma proteins present in human cortical bone and permanent dentine has been determined. One plasma glycoprotein, the alpha2HS-glycoprotein, was found to be present at a high concentration in both bone and dentine and was shown to be concentrated in the mineralized tissues with respect to the other plasma proteins by factors of between 30 and 100. In this respect the alpha2HS-glycoprotein is analogous to the G2B-glycoprotein and alpha-glycoprotein of bovine and rabbit b one, respectively. The binding of alpha2HS-glycoprotein and albumin to calcium phosphates generated within serum samples has been studied at different serum:precipitate ratios. In each case all the alpha2HS-glycoprotein was removed from the samples and the alpha2HS-glycoprotein was concentrated with respect to albumin by factors ranging from 370 at the highest serum:precipitate ratio to 25 at the lowest ratio. The plasma alpha2HS-glycoprotein concentrations of patients with Paget's disease of bone were shown to be substantially lower than the normal range, with significant negative correlation between the alpha2HS-glycoprotein concentration and the plasma alkaline phosphatase activity.


The origin of urinary hydroxylysyl glycosides in Paget's disease of bone and in primary hyperparathyroidism. The urinary excretion of glucosyl-galactosyl-hydroxylysine and of galactosyl-hydroxylysine were studied in Paget's disease of bone and in primary hyperparathyroidism. Both metabolites were increased in these diseases. Although glucosyl-galactosyl-hydroxylysine is not abundant in bone collagen, it is possible that a part of it originates from calcified tissue.


Studies of the calcium accretion rate of bone during immobilization in intact and thyroparathyroidectomized adult rats. In order to study the role played by the thyroid and the parathyroid glands in the development of osteoporosis induced by immobilization, 153 male adult Sprague-Dawley rats allocated in two groups were used. The animals of one group were thyroparathyroidectomized; those of the other group were left intact. Each group was then divided into two subgroups; in one the right hind leg of the animals were immobilized by an elastic adhesive bandage. The animals were observed for varying periods of time up to 16 weeks. Studies of bone mass and 45Ca accretion rate of the right and left femur and tibia were performed in all animals. The results indicate that disuse osteoporosis occurs in rats in the virtual absence of the thyroid and parathyroid glands. The bone loss of the immobilized femur and tibia was less pronounced in thyroparathyroidectomized rats compared to intact rats. The uptake of 45Ca in bone treated by immobilization as compared to the non-immobilized bone was found to be increased to the same proportion in thyroparathyroidectomized and in intact rats. The calcium-accretion rate of non-immobilized bone was lower in thyroparathyroidectomized than in intact rats. The observations indicate that in the rat the parathyroid and thyroid hormones influence the rate of bone turnover and thereby the rate of development of disuse osteoporosis.


Ultrastructural studies on the formation of the periostracum in Helix aspersa (Mollusca). The ultrastructure of the periostracal gland in Helix aspersa and the sequence of formation of the periostracum is described. The periostracal units which measure about 9-12 nm wide and 0.4-0.6 mum long are first detected in the Golgi cisternae. The cisternae containing the unit(s) eventually pinch off from the rest of the Golgi. Microtubules are seen in the space separating the nascent secretory inclusion and the Golgi. Cross-bridges are seen between the microtubules and the secretory inclusions, suggesting that the microtubules are involved in the transport of nascent inclusions away from the site of synthesis. Many periostracal units unite to form periostracal sheets. The mature inclusions containing periostracal sheets migrate to the apical part of the cell where they fuse with the lysosomes before being extruded externally in the lumen of the gland. Perhaps the lysosomal enzymes somehow modify the periostracal units before their extrusion or digest the excess periostracal units. The periostracal sheets released in the lumen disperse randomly to produce a fibrous sheet, which is non-uniform in texture at first but is entirely homogeneous by the time the periostracum is secreted outside.


Plasma fluoride and enamel fluorosis.

Plasma fluoride and enamel fluorosis. It is postulated that tissue fluid F concentrations are the primary determinants of flouride effects on bones and developing teeth and that these concentrations are dependent on, or mirrored by, blood plasma F. It has earlier been shown that the plasma F levels are dependent on the dietary F supply as well as on skeletal F concentration. Fasting and post-ingestion or postinjection plasma F levels have been determined in rats on F doses that cause different degrees of enamel fluorosis. The results indicate that temporary peak values rather than elevated fasting values are responsible for the occurrence of enamel fluorosis and that the peak values must approach about 10 muM in order to block enamel formation by the ameloblasts. The diagnostic and prognostic importance of plasma F determinations is discussed.


Stimulation of bone formation in vivo by phosphate supplementation. The effect of phosphate supplementation on bone remodeling was assessed in six mature, healthy beagle dogs. The phosphate supplement was given in divided doses orally, daily for 12 weeks in the form of a neutral potassium phosphate preparation. The dose averaged 108 mg P/kg per day, which is double the normal canine phosphorus intake. Bone remodeling was assessed by measurement, at sacrifice, of areas of cortical bone containing different color-coded tetracyclines which had been continuously administered during 12-week control and treatment periods; remodeling was assessed kinetically during the control and treatment periods by replicate studies employing 47Ca intravenously. Both techniques demonstrated that the principal effect of phosphate supplementation was a significant stimulation of bone formation. Within cortical bone, formation was doubled, from an average of 2.7% to 5.3% per year. The major location of new bone deposits was endosteal. Whole skeletal mineral accretion, measured kinetically, increased 45% above an average control value of 0.154 g/day. These studies suggest that, in the adult dog, "normal" plasma phosphate levels are suboptimal for new bone formation. Even with this short duration of administration, phosphate produced microscopic calcification of the renal parenchyma. However, there was no biochemical evidence of renal functional impairment.