Electrocardiographic observations on the termination of supraventricular tachycardia by verapamil. The effect of intravenous verapamil on the termination of supraventricular tachycardia (SVT) was studied by continuous electrocardiographic monitoring of 27 episodes of SVT. Progressive increase of the cycle length heralded conversion in eight episodes while cycle-length alternation preceded cessation of the arrhythmia in 13 episodes. In five patients the arrhythmia was either stopped or closely followed by a ventricular premature beat (VPB), followed by further VPBs in three. Runs of bizarre ventricular tachycardia followed initial sinus-beats in two patients. Sinus standstill, lasting 30 seconds, was observed in one patient. The first post SVT beats had an aberrant QRS configuration with a normal P-R interval in four cases and an aberrant QRS complex with a short P-R interval, resembling Wolff-Parkinson-White complexes, in a further seven patients. The possible mechanisms causing this variability of pre- and post-conversion period are discussed. It is suggested that some aspects of verapamil action may be explained by a parasympaticomimetic effect on the myocardium.


Conduction disturbances in patients with calcified mitral annulus diagnosed by echocardiography. Clinical, radiologic and histo-pathological observations have suggested that conduction disturbances (CD) are frequent in patients with calcified mitral annulus (CMA). To determine the incidence of CD, 65 consecutive patients with CMA were studied. Sixty-five normal individuals, matched for age and sex served as a control group. CMA has been identified by M-mode echocardiography, cross-sectional echo and image amplifier fluoroscopy. CD were present in 23 patients with CMA as compared to seven patients in the control group (p less than 0.01). Three patients developed complete atrioventricular block (4.6%) during hospitalization requiring artificial pacemakers. We conclude that, 1) early detection of CMA in the elderly is useful in predicting a higher incidence of CD. 2) Complete heart block is more frequent in patients with CMA. 3) Frequent ECG follow-up in these patients has proven to be useful in avoiding serious clinical complications.


Electrocardiographic QRS changes induced by acute coronary ligation in the isolated rabbit heart. The electrical fields associated with augmented R- and decreased S-wave amplitudes during acute, severe myocardial ischemia were studied in fifteen isolated rabbit heart preparations. Hearts were suspended in a spherical tank and perfused with oxygenated electrolyte solution. Electrocardiographic signals were recorded from electrodes on the tank's surface and processed by computerized methods. Fifteen minutes after suture ligation of the left anterior descending coronary artery, records from electrodes overlying the lesion demonstrated increased R-wave amplitude and ST-segment elevation. Fitting a single moving dipole to pre- and post-ligation potentials demonstrated that the ligation increased the dipolarity of the field and shifted the terminal QRS dipole to a position topographically related to the location of the ischemic lesion. The effects of injury were further assessed by study of fields computed by subtracting control from post-ligation data. This generated a dipolar field at the instant of maximal dipole moment whose strength directly correlated (r=0.74) with the area of non-perfused epicardium, as determined by post-ligation methylene blue infusion. These results suggest that the electrical fields generated during mid to late dipolarization by coronary ligation are qualitatively similar to those generated during repolarization, and may therefore be of similar clinical and/or experimental value.


Effects of stable and changing rates and premature ventricular beats on transient tachycardia-, pseudobradycardia-, and bradycardia-dependent bundle branch block alternans. New circumstances under which bundle block (BBB) alternans may appear or disappear are described. 1) Tachycardia-dependent as well as bradycardia-dependent BBB alternans may begin after constant BBB is interrupted by a premature ventricular beat. Tachycardia- and bradycardia- dependence may be differentiated by the shape of the first beat after the pause. 2) When BBB alternans disappears during a constant ventricular rate, tachycardia-dependent BBB alternans changes to persistent normal or more normal intraventricular conduction, whereas bradycardia-dependent BBB alternans changes to a persistently greater degree of BBB. 3) BBB alternans appears to be tachycardia- or pseudobradycardia-dependent in relation to the cycle length and antegrade and retrograde refractory periods in the involved bundle branch. 4) BBB alternans may be recognized during persistent irregular ventricular action in atrial fibrillation. Here the recognition of BBB alternans depends upon the sequence of contours as well as upon the cycle lengths.


Orthogonal electrocardiographic study on progressive muscular dystrophy of the Duchenne type. An attempt was made to investigate the incidence and significance of high frequency notches and slurs on the QRS complex in patients with progressive muscular dystrophy of the Duchenne type (PMD). The patients were classified into eight stages from the most mild, S(1), to the most severe, S(8), according to Swinyard-Deaver's criteria. Cases where the sum of high frequency notches in the combined leads exceeded none were generally limited to those more advanced than stage S(4). Also, the sum of the notch count tended to be higher in S(5) to S(6) than in the remaining groups and to be lower in the milder cases, S(1) to S(4), and most severe cases, S(7) and S(8). It should be emphasized that cases in groups S(7) and S(8) who had a history of congestive heart failure and/or developed congestive heart failure during the observation period, tended to show a smaller number of notches. High frequency slurs showed almost the same tendencies as the high frequency notches. It is thus anticipated that a significant increase or decrease in the number of notches and slurs on the QRS complex may be suggestive of more intense myocardial derangement.


Predicting coronary artery disease with treadmill stress testing: changes in R-wave amplitude compared with ST segment depression. Coronary angiograms and treadmill stress tests were reviewed independently in 108 nonconsecutively selected cases. There were 16 patients (15%) with infarcts on ECG. Changes in R-wave amplitude and ST segments during exercise were evaluated to determine the sensitivity and specificity of each as a predictor of coronary artery disease (CAD). ST segment changes had a sensitivity of 49%, and a specificity of 74%. The sensitivity increased to 55% when infarcts were excluded. R-wave amplitude changes had a sensitivity of 68% and a specificity of 84%. The sensitivity increased to 78% when infarcts were excluded. An index formed by the sum of the change in R-wave amplitude and the magnitude of ST segment change yielded a sensitivity of 76% and specificity of 78%. The sensitivity increased to 84% when infarcts were excluded. There was no statistical difference between specificities for each criteria. Of those patients with an R-wave amplitude decrease, 69% had no coronary artery atherosclerosis, while 31% had significant lesions. Of those patients with no change or an increase in R-wave amplitude, 83% had coronary artery atherosclerosis, while 17% were normal. Of the 83% with coronary artery atherosclerosis, 81% had two and three vessel disease, while only 19% had single vessel disease. No change or an increase in R-wave amplitude during treadmill stress testing is a more reliable indicator of CAD in our laboratory than ST segment changes.


Body surface potential distributions in posterior ventricular pre-excitation. Waveform of the QRS complex during ventricular pre-excitation is subject to the influence of both the site of pre-excitation and the time of pre-excitation relative to that of excitation via the normal AV path. This paper reports a case in which lead V1 of the electrocardiogram (ECG) could be altered from an R to an rS pattern by the administration of atropine sulfate. The provable mechanism was that of reduced conduction time in the normal AV path with altered time phase of normal excitation and pre-excitation. This mechanism was simulated in experiments on dogs and yielded similar findings. Body surface mapping in both the patient and the dogs provided evidence that pre-excitation could be recognized by that means with varied time phase of normal excitation and pre-excitation. It was demonstrated that the QRS complex of right sided precordial leads could be altered from an R to an rS pattern by altering the time phase of normal excitation and pre-excitation of the posterior ventricular wall. This alteration was related to the degree to which negative potentials on the anterior chest wall due to right ventricular breakthrough of normal activation developed in relation to the time of pre-excitation.


A scale for ECG rhythm analysis.

A scale for ECG rhythm analysis. A simple scale, which allows the ECG reader to assess the regularity of the RR (or PP) interval and to detect the P wave train, has been presented.


Multiple mechanisms of tachycardias in a patient with the Wolff-Parkinson-White syndrome. In a patient with the Wolff-Parkinson-White Syndrome we observed atrial fibrillation and three distinct paroxysmal re-entrant tachycardias. Intracardiac electrograms obtained during the tachycardias showed the mechanisms to be A-V nodal, accessory pathway and sinus node re-entry. When P wave morphology, R-P relationship and QRS configuration are considered, it is illustrated how these four tachyarrhythmias may be successfully diagnosed on the surface electrocardiogram. The therapeutic implications of multiple arrhythmias with different mechanisms in the Wolff-Parkinson-White Syndrome are discussed.


Ebstein's anomaly in association with anomalous nodoventricular conduction. Pre-operative and intra-operative electrophysiological studies. A 13 year old girl with Ebstein's anomaly was investigated for refractory paroxysmal tachycardias and ventricular pre-excitation. Intracardiac electrophysiological studies demonstrated that ventricular pre-excitation was due to conduction in an anomalous nodo-ventricular pathway. Tachycardia occurred as a result of re-entry within the A-V node with pre-excitation during tachycardia due to conduction in the nodo-ventricular pathway. These tachycardias were controlled initially by medical therapy but because of increasing frequency of attacks, occasionally requiring D.C. conversion, further electrophysiological studies and epicardial mapping were undertaken. The epicardial surface of the right ventricle and right atrium were mapped during tachycardia. The results of the studies confirmed that a direct anomalous atrio-ventricular pathway was not present and that re-entrant tachycardia did not involve an accessory pathway of this type. A rapid atrial pacing system was implanted and paroxysmal tachycardias have been successfully controlled.


Plasma testosterone levels in intact and hemicastrated growing cockerels. Changes in the levels of testosterone in plasma were measured by radioimmunoassay in blood samples taken at frequent intervals between 2 and 26 weeks of age from entire cockerels and cockerels hemicastrated before 2 weeks of age. In both groups the pattern of testosterone secretion could be divided into three clearly defined phases. In young birds, the levels of testosterone in plasma were low (0.3 ng/ml) but in the prepubertal period, at 11 weeks of age, they started to rise and continued to rise until 22 weeks of age when adult levels, which fluctuated between 2.5 and 3.5 ng/ml, were reached. In the immediate period after hemicastration, the concentration of testosterone decreased temporarily. From 11 weeks of age the levels of testosterone in the hemicastrated birds were approximately 75% of those in intact birds. These results are discussed in relation to the compensatory testicular hypertrophy which occurs in growing cockerels hemicastrated at an early age.


Inhibition of steroid 5 alpha-reductase in vivo: effect on suppression of luteinizing hormone and stimulation of accessory sex organs by testosterone in the orchidectomized rat. An inhibitor of 5 alpha-reductase, the 17 beta-carboxylic acid derivative of testosterone (testosterone-17 beta CA), has been used to evaluate the importance of the 5 alpha-reduction of testosterone in its action on the suppression of LH secretion in male rats. The potential of testosterone-17 beta CA to inhibit the formation of 5 alpha-dihydrotestosterone (DHT) was first demonstrated in vitro. When homogenates of hypothalami or anterior pituitary glands were incubated with [3H]Testosterone in the presence of a 50-fold excess of testosterone-17 beta CA, the formation of labelled DHT was inhibited by more than 80%. Adult male rats that had been castrated for 1-2 months were fitted with chronic intravenous catheters and implanted with silicone elastomer sheets: one group received one sheet, 0.5-2.0 cm2 in size containing 1.6% testosterone, a second group received one 50 cm2 sheet containing 1.6% testosterone-17 beta CA and a third group received two sheets, one sheet 50 cm2 in size containing 1.6% testosterone-17 beta CA and the second ranging in size from 0.5 to 2.0 cm2 and containing 1.6% testosterone. Blood was withdrawn daily from each rat over a 4-5 day period after implantation of the steroids and the level of LH in the plasma was measured by radioimmunoassay. The seminal vesicles and the ventral prostate gland were removed at autopsy on day 4 or 5; the weights of these organs were shown to have increased progressively as the size of the implant of testosterone increased. In contrast, the level of LH in the plasma was suppressed to a comparable extent by implants of testosterone between 0.6 and 2 cm2, whereas a 0.5 cm2 implant of testosterone had no effect. Implants of testosterone-17 beta CA alone did not influence the weight of the accessory organs or the level of LH. When testosterone-17 beta CA and testosterone were implanted together, the growth-promoting effect of the latter on the accessory sex organs was significantly reduced. The effectiveness of testosterone in suppressing the level of LH in the plasma of these animals was not influenced by the presence of testosterone-17 beta CA and in certain instances the level was raised.


Production of erythropoietic colony-forming units and erythrocytes during chick embryo development: an attempt at modelization of chick embryo erythropoiesis. The enumeration of erythropoietic colony-forming cells in vitro has allowed us to complete previous data on changes in the various erythroid cell populations during chick embryo-genesis. Erythrocytic colony-forming units in culture (CFU-cE) which are sensitive to avian erythropoietin appear in the blastoderm as soon as the 24th hour of development. They represent most likely precursors of the megalocytic erythropoiesis, and do not seem to derive from stem cells common with normocytic erythropoiesis. Data concerning vitelline normocytic erythropoiesis were analysed in a kinetic model based on stochastic change of the stem cells. From this model it appears that 17-20 cell divisions are required for differentiation of erythrocytes from stem cells.


Anomalous ipsilateral optic fibre projection in Xenopus induced by larval tectal ablation. Unilateral left tectal ablation was carried out in Xenopus between stage 48 and 1 month after metamorphosis. Six to 12 weeks after metamorphosis the retinal projection from the right eye was examined with the use of [3H]proline autoradiography. The autoradiographs indicated that optic fibres whose tectal target was destroyed recrossed to the ipsilateral tectum and basal optic nucleus via the posterior and pretectal commissures. No anomalous recrossing occurred if the tectal ablation was carried out at stage 58 or later. The aberrant optic fibres were restricted to the rostrolateral and central parts of the ipsilateral tectum and they terminated in a discontinuous manner. It is concluded that available surfaces serving as contact guidance cues are needed to direct aberrant optic fibres to the ipsilateral tectum.


The cellular effect of 5-bromodeoxyuridine on the mammalian embryo. It is well known that 5-bromodeoxyuridine (BUdR) when injected into pregnant animals may cause exencephaly, cleft palate, and limb abnormalities. Similarly, it is well established that the drug when added to a culture medium may prevent differentiation of embryonic cell systems without affecting cell division or cell viability. The goal of our experiments was to examine whether the congenital malformations resulting from BUdR treatment were due to lack of differentiation of certain cell lines or were due to other mechanisms. The effects of BUdR on proliferating and differentiating cells in the 12-day mouse embryo were therefore examined and special attention was given to the proliferating cells of the rhombic lip which give rise to the Purkinje cells. When the embryos were treated with BUdR the mitotic index of the neuroepithelium of the rhombic lip doubled in value 3 h after treatment and remained high until 24 h later. By using the colchicine index it was calculated that the mitotic duration in the BUdR-treated embryos lasted at least 2 h and that in the control embryos less than 1 h. When the cell generation time in the BUdR treated animals was calculated the length of the S-phase was increased by about 50%. It was thus concluded that BUdR caused an increase in the duration of the S-phase and mitosis, together making the cell cycle 5 h longer than normal. Eighteen hours after treatment many neuroepithelial cells became degenerative. By radioautography it was demonstrated that the degenerating cells were in their second DNA-synthetic phase following BUdR injection and that cells which incorporated BUdR and were differentiating into neurons were not affected. By injecting [3H]BUdR it was found that many cells which incorporated the analogue were able to leave the proliferative population after their first cell division. They migrated to the periphery where they developed into apparently normal Purkinje cells. The additive effects of cell death and retardation of the cell cycle caused a 15% deficit of Purkinje cells in the postnatal cerebellum but the BUdR did not interfere with their differentiation. Thus, contrary to the BUdR effect on cultures of embryonic cells, in vivo the drug causes cell death and a delay in the cell cycle time. Our experiments therefore seem to indicate that the congenital malformations caused by BUdR in the mammalian embryo are caused by cell death and growth retardation rather than by interference with the process of differentiation.


Autoradiographic study of macromolecular synthesis in the fusion epithelium of the developing rat primary palate in vitro. The facial processes involved in primary palate formation undergo epithelial fusion in a manner morphologically analogous to that observed during secondary palate formation. We have used whole embryo culture to analyze the synthesis of macromolecules (DNA, protein glycoprotein) in the primary palate, based on the incorporation of various labeled precursors. The results of this study demonstrate that changes in the synthesis of macromolecules occur during the fusion of the facial processes, which resemble those previously reported to occur during secondary palate development. These changes include cessation of DNA synthesis in cells in a restricted zone of the epithelium, concomitant with maintenance of glycoprotein synthesis. These findings indicate that the molecular events underlying the development of the primary and secondary palate may be similar.


Waves propagated during vertebrate development: observations and comments. The status and possible roles of propagated waves occurring during vertebrate embryogenesis are discussed. Some preliminary data for waves observed in time-lapse films of early chick (Gallus domesticus) and Medaka (Oryzias latipes, a teleost) are given. The general similarities between these phenomena and wave propagation during cellular slime mold aggregation are pointed out and it is suggested that the control of development by propagated waves in regulative embryos is not only widespread but also depends on a common cellular mechanism.


Neurotrophic and x-ray blocks in the blastemal cell cycle. Using microdensitometry techniques the points in the cycle where blastemal cells become blocked after X-irradiation or denervation of the regenerating amphibian limb have been identified. X-irradiation blocks the cells in both G1 and G2 and those cells that were in S at the time of irradiation presumably proceed to G2. After denervation, however, cells accumulate only in G1 and those that were in S or G2 continue through the cycle to the next G1. The latter results are clearly contradictory to a recent theory proposing a G2 neurotrophic control of blastemal cells and a solution to the contradiction is presented in the light of recent results.


The effect of cell killing by x-irradiation on pattern formation in the chick limb. It has been suggested that positional information along the proximo-distal axis of the limb-bud is specified by time spent in the progress zone. Mesenchyme cells have been killed by X-irradiation, reducing the rate cells leave the zone. The time spent there by some cells is thus increased. When limbs, stage 18/19, stage 21, or tips of stage 24, are treated with increasing doses of X-irradiation, from 1000 rads to 2500 rads proximal structures are progressively lost, whereas distal ones--the digits--are relatively unaffected. There was no evidence for intercalation of missing parts. These effects are due to killing or damage of mesenchyme cells: the ectoderm is not affected at these doses. The results are consistent with a quantitative analysis based on the progress zone model, in which viable cells repopulate the progress zone and gradually restore it to normal as non-dividing cells are diluted out. It is suggested that any treatment causing damage to the mesenchyme at early stages will give similar results. The mesenchyme cells appear to be surprisingly resistant to radiation damage. The form of the limb-bud is not altered by damaging the mesenchyme. Differences in the development of structures at similar proximo-distal levels, following irradiation, is considered in terms of the requirement of a threshold number of cells.


The central pathways of optic fibres in Xenopus tadpoles. A cobalt chloride impregnation technique was applied to the optic nerve in Xenopus tadpoles and the central optic pathways were examined in cleared, whole-mounted preparations, and in thick sections. The overall plan of the optic input was visualized in relation to the outlines of the parts of the brain and details of the structure of the tectal optic neuropil, the neuropil of Bellonci and the basal optic neuropil were seen. The fibres in the main retinotectal tract maintained an orderly disposition with respect to each other, in contrast to the fibres of the basal optic tract, in which no order was apparent. Optic fibres were seen passing caudally from the region of the basal optic neuropil.


Effect of the T-mutation on histogenesis of the mouse embryo under the testis capsule. Mouse embryos homozygous for the T-mutation show abnormalities, severer at the posterior embryonic regions, by day 9 of gestation and die before day 11 in utero. To analyse development potentially of the T/T embryos fragments of their anterior and posterior portions were grafted into the testes of adult T/+ mice, and examined histologically for the tissues formed after 1 month. The grafted tissues of the T/T embryos grew beyond the destined lethal stage and gave rise to benign teratomas composed of mature tissues. Although there were some different features of the tissues formed in the teratomas derived from different portions and stages of the embryos, their types were essentially identical between wild-type and the mutant teratomas. Statistical analysis showed that frequency of the cartilage and/or bone formation was significantly lower in the posterior mutant teratomas. It cannot be concluded, however, that this difference is essentially caused by T-mutation. The main conclusion of present experiments is that grafted portions of T/T embryos have the potentiality to develop into teratomas containing derivatives of all three germ layers.


The zone of polarizing activity: evidence for a role in normal chick limb morphogenesis. When an impermeable barrier is placed so as to divide the early chick limb-bud into anterior and posterior parts then development continues only on one side of the barrier. The detailed results are inconsistent with mosaic development. They can readily be explained by supposing that pattern is specified by the concentration of a diffusible morphogen controlled by the zone of polarizing activity. A simulation of appropriate concentration profiles is presented and its relevance to similar experiments published elswhere is discussed. It seems probable that the zone of polarizing activity is active during normal development.


The role of irradiated tissue during pattern formation in the regenerating limb. The amphibian limb regeneration blastema is used here to examine whether irradiated, non-dividing tissue can participate in the development of new pattern of morphogenesis. Irradiated blastemas were rotated 180 degrees on normal stumps and normal blastemas rotated on irradiated stumps. In both cases supernumerary elements developed from the unirradiated tissue. The supernumeraries were defective but this did not seem to be due to a lack of tissue. Rather it suggested that this could be a realization of compartments in vertebrate development or simply reflect the limited regulative ability of the blastema. The results are also discussed in relation to a recent model of pattern formation.


Selection of appropriate medial branch of the optic tract by fibres of ventral retinal origin during development and in regeneration: an autoradiographic study in Xenopus. The formation of the branches of the optic tract has been studied with the use of [3H]proline autoradiography, during development and during regeneration of the optic nerve in Xenopus with one compound ventral (VV) eye made by the embryonic fusion of two ventral eye fragments. The formation of the optic pathway was abnormal in that the lateral branch failed to develop, suggesting that fibres from a VV retina selectively entered the tectum via the medial branch during development. Three months after section of the optic nerve of a VV eye, regenerated fibres were present both in the contralateral and ipsilateral tecta. On the ipsilateral side regenerated fibres entered the tectum via the medial branch only. Retinal fibres entered the contralateral tectum through both branches in some animals and through the medial branch only in others. It is concluded that mechanical factors alone are insufficient to explain the phenomenon of selection of the appropriate medial branch by fibres of ventral retinal origin either during development or in regeneration. Some form of fibre-substrate interaction seems to be necessary; and this ability of fibres from a VV eye to take the path appropriate for ventral retina argues strongly that the VV eye is not a regulated system in terms of cell specificities.


Differentiation capacity of unincubated chick blastoderm in culture. Unincubated (stage X) chick blastoderms cultured in plain Dulbecco's modified Eagle medium in the absence of serum, synthesize haemoglobin and melanin 2 . 5--3 . 0 and 6 . 0--7 . 0 days, respectively, after the beginning of culture. The capacity of unincubated blastoderm to form haemoglobin and melanin is a response to intrinsic genotypic instructions rather than to the active intervention of substances continuously supplied by the yolk. 5-Bromodeoxyridine, applied during the first and second days of unincubated blastoderm in culture, is incorporated into DNA and perturbs the formation of haemoglobin and melanin, perhaps, by modifying gene expression.


The ontogeny of erythropoiesis in the mouse detected by the erythroid colony-forming technique. I. Hepatic and maternal erythropoiesis. Employing the erythroid colony-forming technique, it is shown that throughout hepatic erythropoiesis in the mouse, the CFU-E population remains sensitive to erythropoietin. Maximum stimulation was achieved during this period using an erythropoietin concentration of 0.075 units/ml. The peak in the CFU-E concentration occurs between the 11th and 12th day while absolute values show a maximum on the 14th day of gestation. These results are discussed in terms of changing cell populations, both of erythropoietic precursors and hepatocytes from which it is concluded that at no time during foetal erythropoiesis does the CFU-E population change or become unresponsive to erythropoietin. The BFU-E population follows closely that of the CFU-E, but declines about 24 h earlier on the 16th day of gestation. The effect of the foetus on the mother was also studied during the second half of pregnancy. During this period of natural perturbation both femoral and, in particular, splenic erythropoiesis are increased. However, during this time an erythropoietin concentration of 0.3 units/ml was required to maximally stimulate the CFU-E population derived from these tissues. The fact that both adult and foetal erythroid tissue maintain a rather constant requirement for erythropoietin for their growth in vitro, indicates that it is an intrinsic property of the cells. It is concluded that increased maternal erythropoiesis is due to an increased oxygen requirement causing hypoxia due to the growing foetus.


In vitro studies on the morphogenesis and differentiation of the mesoderm subjacent to the apical ectodermal ridge of the embryonic chick limb-bud. It has been suggested that one of the major functions of the apical ectodermal ridge (AER) of the embryonic chick limb-bud is to maintain mesenchymal cells directly subjacent to it (i.e. cells extending 0.4-- 0.5 mm from the AER) in a labile, undifferentiated condition. We have attempted to directly test this hypothesis by subjecting the undiffertiated subridge mesoderm of stage-25 embryonic chick wing-buds to organ culture in the presence and absence of the AER and the ectoderm that normally surrounds the mesoderm dorsally and ventrally. During the period of culture, control explants comprised of the subridge mesoderm capped by the AER and surrounded by the dorsal/ventral ectoderm undergo progressive morphogenesis characterized by polarized proximal to distal outgrowth and changes in the contour of the developing explant, and ultimately form a structure grossly resembling a normal distal wing-bud tip. In contrast, explants from which the AER and dorsal/ventral ectoderm have been removed (minus ectoderm explants) or from which just the AER has been removed (minus AER explants) form compact, rounded masses exhibiting no signs of morphogenesis. During the polarized proximal to distal outgrowth control explants undergo during the first 3 days of culture, as cells of the explant become located greater than 0.4--0.5 mm from the AER, they concomitantly undergo a sequence of changes indicative of their differentiation into cartilage. However, those cells which remain 0.4--0.5 mm from the AER during this period retain the characteristics of non-specialized mesenchymal cells. In marked contrast to control explants, virtually all of the cells of minus ectoderm explants initiate chondrogenic differentiation during the first day of culture. Cells comprising the central core of minus AER explants also initiate chondrogenic differentiation during the first day of culture, but in contrast to minus ecotderm explants, non-chondrogenic tissue types form along the periphery of the explants subjacent to the dorsal/ventral ectoderm. These results indicate that the AER maintains cells directly subjacent to it in a labile, undifferentiated condition, and that when mesenchymal cells are freed from the AER's influence either artificially or as a result of normal polarized outgrowth, they are freed to commence cytodifferentiation. The results further suggest that the dorsal/ventral ectoderm may have an influence on the differentiation of the mesenchymal cells directly subjacent to it, once the cells have been removed from the influence of the AER.


Inhibition of tooth germ differentiation in vitro by diazo-oxo-norleucine (DON). Molar tooth germs from mouse embryos were studied in a Trowell-type organ culture. After 5 days of culture the odontoblasts had secreted predentine and the ameloblasts had differentiated. When cultured in the presence of 10-50 micro M diazo-oxo-norleucine (DON), which is a glutamine analogue, the differentiation of odontoblasts was inhibited, but the teeth looked otherwise healthy. When DON was added after 2 days of culture in control medium (at this time the odontoblasts in the cuspal area were already differentiated), it did not inhibit predentine secretion, ameloblast differentiation, nor enamel secretion. However, this was seen only in the cuspal area and the boundary to the undifferentiated, more cervical cells was distinct. The results support the concept that the mechanism of the differentiation of odontoblasts is different from that of the ameloblasts. We have shown earlier that a close association between the basement membrane and the mesenchymal cells is required for odontoblast differentiation. Because DON interferes with glycosaminoglycan and glycoprotein synthesis we suggest that DON inhibits odontoblast differentiation by affecting the mesenchymal cell surface and/or the basement membrane.


Regional blood flow and its relationship to lymphocyte and lymphoblast traffic during a primary immune reaction. The relationship of alterations in blood flow with changes in cell distribution has been studied in an inflammatory site and its draining lymph node during the induction of an immune reaction with oxazolone in mice. The cells which move to the site of inflammation are predominantly lymphoblasts and their increased localization in the inflamed ear is significantly correlated with increased regional blood flow to the inflamed tissue. The existence of this correlation is not antigen dependent although there is a relative increment of lymphoblasts which are specifically primed to the inflammatory agent. The localization of nonblastic (small) 51Cr-labeled lymphocytes on the other hand is substantial only in lymphoid tissue and during the induction of an immune reaction after oxazolone application, the increase in localization of these cells in the draining lymph node is positively correlated with increased blood flow to the node. Furthermore, the probability of finding 51Cr labeled lymphocytes in a particular lymph node is related to the regional blood flow which that node receives.


The requirement for adherent cells in the Fc fragment-induced proliferative response of murine spleen cells. The proliferative response of mouse B lymphocytes induced by Fc fragments was found to be dependent upon an adherent cell population. The adherent cell is esterase positive, irradiation resistant, and not susceptible to lysis by anti-thymus serum and complement. The mechanism(s) by which Fc fragments induce B-cell proliferation could be the result of the interaction of Fc with both B cells and adherent cells or with adherent cells which then release factors that trigger the B cells to proliferate. Spleen cells from the C3H/HeJ mouse were shown to be unable to respond to Fc fragments. The addition of adherent cells from either C3H/St or C3H/HeN mice to adherent cell depleted C3H/HeJ cells enabled them to respond to Fc, indicating the defect was in the adherent cell population.


The role of C4-binding protein and beta 1H in proteolysis of C4b and C3b. Two forms of C4-binding protein (C4-bp) (C4-bp low, C4-bp high), which differ slightly in net charge and apparent molecular weight, as determined by SDS- PAGE, were separated by ion-exchange chromatography and contaminants removed with specific antisera. Both forms of C4-bp served as cofactors for the cleavage of C4b in solution by C3b inactivator, and the resulting fragments of the a'-chain of C4b had identical molecular weights. In addition, similarly to beta1H, C4-bp low or high served as cofactors for the cleavage of fluid phase C3b by C3bINA. However, important quantitative differences between the activities of C4-bp and beta1H were observed. With regard to C3b in solution, the cofactor activity of beta1H was {approximately equal to}20 times greater than that of C4-bp on a weight basis. In relation to cell-bound C3b, the differences in activity were even more marked. Whereas beta1H enhanced the effects of C3bINA on the erythrocyte intermediate EC3b, inhibiting the assembly of EC3bBb, C4-bp was without effect even at concentrations {approximately equal to}300 times greater than beta1H. Therefore, under physiological conditions, it is likely that beta1H is the key protein which controls the function of C3b, and that C4-bp activity is directed mainly toward the cleavage of C4b. We also examined the relation between C4-bp and the C3b-C4bINA cofactor described by Stroud and collaborators (3, 4). By functional, physico-chemical and immunological criteria, they are the same protein.


Monocytes regulate osteoclast-activating factor production by releasing prostaglandins. Osteoclast-activating factor (OAF), a powerful stimulator of osteoclastic bone resorption, is released by peripheral blood mononuclear cells on exposure to phytohemagglutinin (PHA) or a specific antigen to which the leukocytes have been previously exposed. Both lymphocytes and monocytes are required in the leukocyte population for OAF release to occur. In this study we examined the relationship between the lymphocyte and monocyte in OAF production. Biological activity, as a result of OAF, was assessed by a bioassay based on the release of previously incorporated 45Ca from fetal rodent long bones in organ culture. We found that an enriched lymphocyte population depleted of monocytes by serial adherence does not release OAF after stimulation with PHA, although the cells are activated as assessed by [3H]thymidine and 3H-amino acid incorporation. When conditioned media harvested from adherent cells which did not contain OAF was added to the enriched lymphocytes, OAF release occurred. Media harvested from adherent cells which were cultured with indomethacin (10 microM), an inhibitor of prostaglandin synthesis, did not permit OAF release by activated lymphocytes. When PGE1 and PGE2 (0.1 microM) were added exogenously to the enriched lymphocyte population, OAF release occurred after stimulation with PHA. These results indicate that, (a) the activated lymphocyte is the cell or origin of OAF, (b) prostaglandins produced by monocytes are necessary for OAF production by activated lymphocytes, and (c) monocyte prostaglandins can influence bone resorption indirectly by regulating OAF production as well as directly by osteoclast activation. The interactions of OAF and prostaglandins at bone resorbing sites may be important in inflammatory and neoplastic diseases associated with bone destruction.


Radioimmune analysis of an unexpected mouse allotype. We have used a radioimmune assay to confirm our earlier findings of an unexpected immunoglobulin allotype in Igb-congenic BALB/c mice. Although these mice were bred to exclude the IgG2a allotype of BALB/c (Ig-la), an Ig-la-like antigen was detected in the 7S Ig fraction of two (of five) pooled serum samples, it represented 0.1--0.3% of the total 7S protein and was indistinguishable from a reference Ig-la. The detection of putative Ig-la in Igb-congenic mice is inconsistent with the notion that allotypes are products of allelic structural genes. It appears rather that expression of Ig-la is controlled by allelic regulator genes and that its low and transient production in Igb-congenic mice results from incomplete negative regulation.


Studies on the mechanisms of macrophage activation. II. Parasite destruction in macrophages activated by supernates from concanavalin A-stimulated lymphocytes. Activation of mouse peritoneal exudate macrophages, as evidenced by destruction of the intracellular protozoan parasite Leishmania enriettii, was obtained by incubation with supernates from concanavalin A (Con A)-stimulated syngeneic spleen cells. Parasites were not destroyed in macrophages exposed to control media. Supernate-induced activation was independent of the presence of Con A. The activating principle (macrophage activating factor, or MAF) was produced by Con A-stimulated lymphocytes in presence or absence of serum. In absence of serum, MAF synthesis was highest at Con A concentrations far below those required in serum-containing media. MAF production was reduced at Con A concentrations of 10 microgram/ml or above, probably a result of toxicity of the lectin for lymphocytes. MAF was detectable after 24 h of lymphocyte stimulation and increased up to 72 h; production appeared to be independent of DNA synthesis. Serum-free MAF was inactive when tested as such on macrophages. Full activity could be restored by addition of nanogram amounts of endotoxin or of FCS before assay. Endotoxin also considerably potentiated MAF activity in serum-containing supernates. Full intracellular parasite destruction was observed after contact of macrophages with MAF for 20 h. The continuous presence of MAF was not necessary for activation; a 10-h pulse was sufficient to induce macrophages to destroy all intracellular microorganisms within the next 38 h.


Generation of unique mono-hydroxy-eicosatetraenoic acids from arachidonic acid by human neutrophils. Incubation of [3H]arachidonic acid with the 17,000-g supernatant from homogenates of human neutrophils in the presence of indomethacin generated the unique metabolites 9-OH-5,7,11,14-eicosatetraenoic acid (9-HETE) and 8-HETE, in addition to 12-HETE, 11-HETE and 5-HETE. The human neutrophil chemotactic activity of the HETE products exhibited a rank-order of potency with 5-HETE greater than 8-HETE = 9-HETE greater than 11-HETE = 12-HETE. The expression of chemokinetic activity as well as chemotactic activity suggested that the endogenous production of these principles may influence the mobility of human neutrophils.


A murine teratocarcinoma stem cell line carries suppressed oncogenic virus genomes. Murine teratocarcinoma stem cells are nonpermissive for productive infection by a variety of DNA (polyoma and SV40 virus) and RNA (murine leukemia and sarcoma virus) tumor viruses whereas differentiated murine cells derived from the stem cells are permissive for productive (or abortive in the case of SV40) infection by these same viruses. The block to productive infection by these oncogenic viruses is at a postpenetration step in the replication cycle of these viruses but the precise level of the block has not been established for any of these viruses. In this report we describe teratocarcinoma-derived stem and differentiated cell lines which should be especially useful in determining the level of the block to replication of ecotropic murine leukemia virus in murine teratocarcinoma stem cells. The stem cell line, OTT6050AF1 BrdU, which is completely nonpermissive to productive infection by Moloney murine leukemia virus and consists of 97% pluripotent stem cells, contains DNA copies of an RNA tumor virus which is indistinguishable from the N-tropic murine leukemia virus of AKR mice. The stem cells are negative for expression of viral reverse transcriptase, p30 and gp69/71 and no virus is found by XC plaque assay or other biological tests. Differentiated cells established from the same teratocarcinoma tumor are 100% positive for viral gp69/71, p30, and produce large amounts of reverse transcriptase activity and N-tropic virus as detected by biological assay. The virus isolated from the differentiated cells is closely related, if not identical to AKR N-tropic virus by nucleic acid hybridization studies and is thus not an endogenous virus of the 129 strain of mice. The teratocarcinoma tumor from which the cell lines were established had been carried in 129 mice and perhaps at some time in the mouse passage history the tumors were infected (nonproductively) with the N-tropic virus. Regardless of the origin of this viral DNA, the OTT6050A derived stem and differentiated cell lines should be extremely useful in defining in stem cells the step at which ecotropic murine leukemia virus replication is blocked.


Incorporation of [U-14 C]glucose into metabolites of brain, liver and blood of rats pretreated with reserpine or phenothiazines. Parkinsonism was induced in rats by using phenothiazines (Butyrylperazin and Thioproperazin). (P-group), or reserpine, (R-group). [U-14 C)D-glucose was administered when the symptoms of Parkinsonism had become fully developed. Concentrations and radioactivities of different metabolites were studied in brain, liver and blood serum. 1. Both types of treatments resulted in a decrease in the synthesis of amino acids from [14C]glucose in the brain. The concentrations of amino acids and the glycogen remained uneffected. Phenothiazines enhanced the conversion of lipids, while reserpine increased their concentration. 2. Reduced de novo synthesis of amino acids was recorded in the liver. Phenothiazines resulted in the storage of glycogen and lipids; reserpine resulted in the storage of lipids and enhanced the conversion of glycogen. 3. Both treatments caused a fall in the amino acid concentration of the blood serum. A rise in the specific radioactivity of blood amino acids was observed in the P-group, while a decrease in specific radioactivity was observed in the R-group. A hyperglycemia was induced in the R-group with reduced specific radioactivity of glucose in both P-and R-groups. A reduction in lipid concentration of blood serum was achieved with an increased specific radioactivity in P-group and decreased radioactivity in R-group. 4. The changes in amino acids common to both treatments are also observed in human Parkinsonism.


[The activity of the peroxide-metabolizing system in human liver (author's transl)]. The activities of various peroxide-metabolizing enzymes were determined in homogenates of human liver excisions. The specific activity of selenium-dependent glutathione peroxidase was 41.1 +/- 23.7 (S.D.) mU/mg protein; non-selenium glutathione peroxidase showed a activity of 30.5 +/- 14.0 mU/mg protein. The catalase and superoxide dismutase concentrations were 4.72 +/- 0.58 and 1.87 +/- 0.68 microgram/mg protein, respectively. Total glutathione amounted to 12.9 +/- 7.4 nmol/mg. Malondialdehyde formation, used as the basis for the determination of lipid hydroperoxides, was 0.32 +/- 0.14 nmol/mg. The data indicate much lower enzyme and substrate levels compared to rats and mice. A positive correlation of r = 0.48 +/- 0.31 was found between the glutathione level and selenium-dependent peroxidase. Selenium-dependent and non-selenium-glutathione peroxidase correlate negatively (r = -0.71 +/- 0.18); superoxide dismutase concentration and lipid-hydroperoxides are also related by a negative correlation coefficient of r = 0.47 +/- 0.31. These data stress the major hepatoprotective role of these systems in human liver.


Aids to the evaluation of diagnostic performance, as applied to immunological creatine kinase-MB assay in myocardial infarction. Immunoassay of creatine kinase-MB provides numerical information, which makes it possible to estimate quantitatively the diagnostic performance following myocardial infarction. Two graphical methods for such an evaluation are presented. The relationship between technical sensitivity and specificity was analyzed using a continuous function, the receiver-operator characteristic curve. Using this function, a diagnostic threshold ("upper normal limit") was chosen, which balances both technical specificity and sensitivity on the first day following infarction. On the second and third days this threshold caused a progressive loss of technical sensitivity. The relationship between effectiveness and the prevalence of myocardial infarction in the tested population was evaluated with a nomogram correlating these two quantities. With the chosen diagnostic threshold, effectiveness is independent of prevalence on the first day, but the loss in technical sensitivity on subsequent days causes effectiveness to decay when the prevalence is high.


[The use of reptilase for electrophoresis of heparinized plasma (author's transl)]. Fibrinogen can be precipitated from Heparin-plasma by Reptilase (Batroxobin). The electrophoretic pattern of the supernatant on cellulose acetate is nearly identical with that of the corresponding serum. This technique is also useful in differentiating a gradient of fibrinogen (caused by effects of anticoagulants) from a myeloma peak.


Evaluation of methods for the estimation of 5-aminolevulinate dehydratase for a broad range of lead concentrations in the blood of exposed workers. 5-aminolevulinate dehydratase activity was estimated in the blood of 30 lead-exposed workers and 15 control persons, using (I) the method of Tomokuni ((1974) Arch. Environm. Health 29, 274--281) and (II) the European standard method (Berlin & Schaller (1974) Z. Klin. Chem. Klin. Biochem. 12, 389--390). The lead level in the blood was in the range 0--11.6 mumol/l. It was found that the correlation between the activity of 5-aminolevulinate dehydratase and the lead level in blood (up to 4.35 mumol/l) are higher for method I than method II. These two methods only give similar results for the units of 5-aminolevulinate dehydratase activity when the lead concentrations in the blood are low. For higher concentrations of this metal (up to 11.6 mumol/l), a high correlation was obtained (r = -- 0.80) between the 5-aminolevulinate dehydratase activity ratio (enzyme measured at pH 6.8/enzyme measured at pH 6.0; method I) and lead concentrations in the blood.


Comparison of acidic and basic chromosomal proteins from normal human endometrium and undifferentiated endometrial carcinoma by isoelectric focussing and microgel-electrophoresis. In the study reported here, non-histamine chromosomal proteins from proliferative and secretory human endometrium, and from undifferentiated endometrial carcinoma have been separated into more than 750 protein components, using a new preparative and highly sensitive analytical method. The following experimental procedure was applied: 1. Dissociation of chromatin under different conditions (variable parameters: ion strength, dissociation agents, shearing, pH), 2. cation exchange chromatography over Bio Rex, 3. preparative fractionation of those non-histamine chromosomal proteins which are not adsorbed on Bio Rex 70 in a Valmet-electrofocusing apparatus, 4. micro-electrophoresis of the focused proteins in microgels containing a continous gradient of polyacrylamide, 5. two-dimensional electrophoresis of the strongly basic chromosomal proteins. There are qualitative differences with respect to the components of this class of proteins between proliferative and secretory endometrium and endometrial carcinoma. The relevance of these results to the control of gene activity is discussed.


Cow's milk allergy: a critical review.

Cow's milk allergy: a critical review. Foods which are ingested frequently, and cow's milk in particular, are a common cause of delayed-in-onset allergy. Difficulty in diagnosing milk allergy is encountered because: (1) skin tests are unreliable; (2) the symptoms of milk allergy are frequently delayed in appearing, thereby obscuring their connection with the previously ingested food; (3) when such symptoms do appear, they often fail to suggest allergy as etiologic; and (4) the offending antigen may be present in a milk product not obviously associated with milk per se such as sherbet or cheese. Contrary to common belief, most milk (and other food) allergy is not reagin (IgE) mediated. Milk allergy is not confined to infancy, but is frequently seen in children and adults, often persisting as an allergy which had supposedly been "outgrown." Since milk allergy is often a familial disorder, its presence in a patient can assist the physician in possibly discovering it in other members of the family. A two or three-week trial elimination of milk, in conjunction with written dietary instructions for the patient, is presently the only reliable diagnostic procedure.


Podophyllum toxicity: case report and literature review. A case is reported of apparent podophyllum toxicity. The patient was a 17-year-old female Indian who had received some 3 to 8 cc of a 20 percent mixture of podophyllum resin in compound tincture of benzoin (approximately equal to 0.4 gm of podophylotoxin) as an application to her vulvar condylomata. She returned to the hospital 20 hours after application in a comatose state. On examination she demonstrated severe toxicity including bone marrow, liver, and central nervous system. She required a charcoal hemoperfusion at the University of Colorado, and six months after the event has several neurologic and physiologic sequelae. Podophyllum is a potentially severely toxic drug. Great care must be taken when treating patients with this drug. A large mass of condylomata or the status of pregnancy should be relative contraindications to the use of podophyllum.


Teaching attitudes and skills in microcounseling. This is the first of two papers describing salient aspects of the behavioral science orientation curriculum in the Charleston Family Practice Residency that focus on developing counseling skills. Two short lectures preceding microcounseling training are summarized. One short lecture outlines the myth of medical management and differentiates a "problem oriented" from a "client oriented" model. The second short lecture presents the definition of communication as a culturally structured complementary series of behaviors. Microcounseling, which is an experientially oriented series of communication exercises, is presented in detail. The article concludes with a discussion of the specific advantages and disadvantages of this particular learning format and its implications for the practice of family medicine.


Observations and impressions of the Canadian health care system. The United States government, in its desire to deliver broad health care coverage to its citizens, has looked to several of the established socialized health care systems for direction. There are definitely good points in each system, and the Canadian system, in particular, has done quite well in providing services within a limited federal budget. On the other hand, the unlimited access to care has led to increased demands for health care services, overperformance of services, and excessive utilization of facilities. There are major technological constraints now emerging and the fiscal integrity of the system is shaky. There is a notable decrease in research and voluntary faculty participation at university levels. Financial constraints are becoming more severe and it appears that demand vis-a-vis the resources available will soon force stringent readjustments in Canadian health care delivery and funding. Health care plan administrators concede that unless more dollars are invested in the system, the current level of health care delivery cannot be maintained.


Morbidity in the first year of a family practice and its comparison to the Virginia study. In the first year of practice two family physicians recorded 8,795 new problems. The problems were ranked in order of decreasing frequency and compared to the results of the Virginia study. The results support the concept that a curriculum for family practice residency training based on results of the Virginia study is a valuable concept. The study also sheds light on the patterns of disease in a new practive as well as suggesting applications for the use of the disease index in practice management.


The agitated patient.

The agitated patient. The emergency management of the agitated patient is a common medical problem. Agitated behavior is not a diagnosis but a descriptive term; the initial task of the physician involves determining the etiology of the behavioral disturbance and evaluating the possible contribution of organic factors. Such factors as age of onset, acuteness of onset, concurrent illness, evidence of delirium or dementia, or use of exogenous pharmacologic agents require careful evaluation. Agitated patients will generally fall into one of four diagnostic categories: agitation precipitated by drug intoxication, agitation precipitated by drug withdrawal, agitation precipitated by an organic brain syndrome, or agitation precipitated by a functional disorder. Appropriate pharmacological and psychological management techniques for these situations are discussed.


The contribution of a sensitizing pigment to the photosensitivity spectra of fly rhodopsin and metarhodopsin. Most of the photoreceptors of the fly compound eye have high sensitivity in the ultraviolet (UV) as well as in the visible spectral range. This UV sensitivity arises from a photostable pigment that acts as a sensitizer for rhodopsin. Because the sensitizing pigment cannot be bleached, the classical determination of the photosensitivity spectrum from measurements of the difference spectrum of the pigment cannot be applied. We therefore used a new method to determine the photosensitivity spectra of rhodopsin and metarhodopsin in the UV spectral range. The method is based on the fact that the invertebrate visual pigment is a bistable one, in which rhodopsin and metarhodopsin are photointerconvertible. The pigment changes were measured by a fast electrical potential, called the M potential, which arises from activation of metarhodopsin. We first established the use of the M potential as a reliable measure of the visual pigment changes in the fly. We then calculated the photosensitivity spectrum of rhodopsin and metarhodopsin by using two kinds of experimentally measured spectra: the relaxation and the photoequilibrium spectra. The relaxation spectrum represents the wavelength dependence of the rate of approach of the pigment molecules to photoequilibrium. This spectrum is the weighted sum of the photosensitivity spectra of rhodopsin and metarhodopsin. The photoequilibrium spectrum measures the fraction of metarhodopsin (or rhodopsin) in photoequilibrium which is reached in the steady state for application of various wavelengths of light. By using this method we found that, although the photosensitivity spectra of rhodopsin and metarhodopsin are very different in the visible, they show strict coincidence in the UV region. This observation indicates that the photostable pigment acts as a sensitizer for both rhodopsin as well as metarhodopsin.


The gecko visual pigments. The behavior of opsin. The 521-pigment extracted out of the retina of the Tokay gecko has the typical stereospecificity of the vertebrate visual pigments. This is true for the pigment in the chloride-depleted, "blue-shifted" state as well as for the normal pigment with added chloride. While in the chloride-deficient state, pigment regeneration occurred with both 11-cis- and 9-cis-retinals and the regenerated photopigments were also in the blue-shifted, chloride-depleted state. As with the native pigment, these regenerated pigments were bathochromically shifted to their normal positions by the addition of chloride. Chloride-deficient opsin by itself also responded to chloride for the pigment regenerated with 11-cis-retinal from such chloride-treated opsin was in the normal 521-position. Regeneration was always rapid, reaching completion in less than 5 min, and was significantly faster than for cow rhodopsin regenerating under the same conditions. This rapid rate was found with or without chloride, with both 11-cis- and 9-cis-retinals and in the presence of the sulfhydryl poison, p-hydroxymercuribenzoate (PMB). Like the native chloride-deficient pigment, the regenerated chloride-depleted photopigments responded to PMB by a blue shift beyond the position of the chloride-deficient state. The addition of chloride to these "poisoned" regenerated pigments caused a bathochromic shift of such magnitude as to indicate a repair of both the PMB and chloride-deficient blue shift. In this discussion the possible implications of these results to phylogenetic considerations are considered as well as to some molecular properties of the 521-pigment.


Sodium deprivation alters neural responses to gustatory stimuli. The effects of sodium deprivation for 10 d, a period sufficient to induce sodium appetite, on gustatory nerve discharges in rats were determined. Chorda tympani responses to concentration series of sodium chloride, sucrose, hydrochloric acid, and quinine hydrochloride were recorded and analyzed without the experimenter knowing the animal's deprivation condition. After deprivation, both whole nerve and single nerve fiber responses to sodium chloride were smaller; NaCl-best fibers, those more responsive to sodium chloride than to sucrose, hydrochloric acid, or quinine, were most affected. Thresholds had not changed; however, slopes of the stimulus-response functions for sodium chloride were lowered. Comparable changes in responses to the other stimuli did not occur. These results were discussed with respect to a possible relationship between changes in sodium chloride responsivity and changes in sodium intake, differences between methods of inducing sodium appetite, coding of taste quality and intensity, and mechanisms which might effect the responsivity change.


Propagation of action potentials in squid giant axons. Repetitive firing at regions of membrane inhomogeneities. Effects of reduction in potassium conductance on impulse conduction were studied in squid giant axons. Internal perfusion of axons with tetraethylammonium (TEA) ions reduces G K and causes the duration of action potential to be increased up to 300 ms. This prolongation of action potentials does not change their conduction velocity. The shape of these propagating action potentials is similar to membrane action potentials in TEA. Axons with regions of differing membrane potassium conductances are obtained by perfusing the axon trunk and one of its two main branches with TEA after the second branch has been filled with normal perfusing solution. Although the latter is initially free of TEA, this ion diffuses in slowly. Up until a large amount of TEA has diffused into the second branch, action potentials in the two branches have very different durations. During this period, membrane regions with prolonged action potentials are a source of depolarizing current for the other, and repetitive activity may be initiated at transitional regions. After a single stimulus in either axon region, interactions between action potentials of different durations usually led to rebound, or a short burst, of action potentials. Complex interactions between two axon regions whose action potentials have different durations resembles electric activity recorded during some cardiac arrhythmias.


Phenoxazinone biosynthesis: accumulation of a precursor, 4-methyl-3-hydroxyanthranilic acid, by mutants of Streptomyces parvulus. Mutants of Streptomyces parvulus that are blocked in the synthesis of the phenoxazinone-containing antibiotic, actinomycin, were isolated by the 'agar piece' method (after ultraviolet irradiation or treatment with 8-methoxypsoralen plus near-ultraviolet light). Radiolabelling experiments in conjunction with paper, thin-layer and column chromatography revealed that 4-methyl-3-hydroxyanthranilic acid (MHA) is a major metabolite accumulated by these mutants. Studies in vitro and in vivo provided evidence that MHA is a precursor of the phenoxazinone chromophore, actinocin. Normally MHA does not accumulate during growth or antibiotic synthesis by the parental strains. Protoplasts derived from the mutant strain AM5 synthesized MHA in significant amounts. A scheme is proposed for the biosynthesis of actinomycin D that accounts for the accumulation of MHA by the mutants.


Factors affecting recombinant frequency in protoplast fusions of Streptomyces coelicolor. The optimum concentration of polyethylene glycol 1000 (PEG) for the production of recombinants through protoplast fusion in Streptomyces coelicolor was about 50% (w/v). The addition of 14% (v/v) dimethyl sulphoxide to the fusion mixture enhanced recombination frequencies, but only at sub-optimal PEG concentrations. After treatment of protoplasts with 50% PEG for 1 min, the frequency of recombinants in a multi-factor 'cross' sometimes exceeded 20% of the total progeny. The frequency of recombinants in the progeny could be significantly enhanced by ultraviolet irradiation of the parental protoplast suspensions immediately before fusion.


Modulation by centrifugation of cell susceptibility to chlamydial infection. Enhancement of chlamydial infection of cell monolayers by centrifugation was shown to depend on induced cell surface changes. Evidence for this came from analysis of two forms of organism attachment which take place during centrifugation. In 'productive binding', organisms attached to cells and then entered and infected them. In 'unproductive binding', organisms became attached to cells but were not ingested. These organisms could be stripped from the cells by treatment with trypsin and could then infect fresh monolayers. Measurement of attachment kinetics during centrifugation showed that cells passed through three different susceptibility states. Only productive binding occurred in the first 20 min; cells then entered a refractory state during which no attachment took place At about 45 min, attachment recommenced but this allowed only unproductive binding. Induced movement of cell surface structures may enhance infection by promoting specific or non-specific interactions. Failure of ingestion may result from insufficient cell 'receptors' for circumferential binding of the whole chlamydial surface so that engulfment cannot take place.


Object perception and motion in infants. Male and female infants 10 weeks of age (N = 24) were presented with two distinctively patterned objects in either moving or stationary form. Measures of visual regard indicated the infants were able to discriminate between the objects whether the objects were stationary or in motion. The results are contrasted to T. G. R. Bower's assessment of infants' utilization of features in perceiving moving objects. The results are also compared to those of an earlier study which employed an operant rather than a preference technique to investigate this same problem.


The effects of stomach loads of hypertonic NaCl on thyroidectomized prepubescent rats. Prepubescent male and female rats (N = 56) were surgically thyroidectomized and given stomach loads of .9% or 6.0% NaCl. The 6.0% NaCl, thyroidectomized S s responded by decreasing food intake and losing weight 24 hours after loading. Normal S s did not, and no sex difference was found. The .9% NaCl thyroidectomized S s drank significantly less than the normals. The results were interpreted in terms of thyroid hormone activity which affects renal function.


Behavioral and neuroendocrine effects of low dose ET-495: antagonism by haloperidol. Low doses of the dopamine agonist ET-495 were administered to nonpsychotic volunteer subjects by slow intravenous infusion, followed by a bolus of 1.5--2.5 mg haloperidol. ET-495 caused progressive dysphoria and sedation (in some cases, light sleep), effects believed to be mediated by dopaminergic inhibition. However, ET-495 also elevated growth hormone and suppressed prolactin, typical responses to dopamine agonist activity. Haloperidol reversed both the sedation and prolactin suppression induced by ET-495. These findings suggest: (1) that the sedation and hormonal responses were produced by stimulation of dopamine receptors; (2) that neurotransmitter systems mediating behavioral and neuroendocrine regulation may have differential neuropharmacological characteristics.


In vitro studies on the possible effects of 1-aminoadamantanes on the serotonergic system in M. Parkinson. Synaptosomes, synaptic vesicles, and membranes were isolated from rat brain homogenates by differential and density gradient centrifugation in sucrose. Synaptosomes incorporated serotonin (5-HT) with two different uptake mechanisms, high affinity: Kt1 = 47 nM and low affinity: Kt2 = 660 nM. Both uptake mechanisms are non-competitively inhibited by the potential antiparkinson drugs 1-aminoadamantane (amantadine, D 1: Ki1 = 57 microM, Ki2 = 96 microM) and 1-amino-3,5-dimethyladamantane (memantine, D 145: Ki1 = 97 microM, Ki2 = 150 microM). The incorporated 5-HT is released from synaptosomes on incubation with high concentrations (0.5--5 mM) of the drugs or on electrical stimulation with rectangular pulses of alternating polarity. Subthreshold concentrations of these drugs (5--50 microM) which are too low to liberate 5-HT increase the electrically stimulated release of 5-HT.--The effect of D 1, D 145, and electrical stimulation on DA release parallels the results observed with 5-HT. The uptake of 5-HT into isolated synaptic vesicles and the binding to isolated nerve ending membranes is non-competitively inhibited by 1-aminoadamantanes. D 145 inhibits the binding of 5-HT to membranes more effectively (Ki = 0.95 mM) than its uptake into vesicles (Ki = 1.2 mM) contrasting with D 1 which is a weaker inhibitor affecting vesicular uptake (Ki = 2.5 mM) slightly more than membrane binding (Ki = 3.1 mM). The results obtained suggest that, in addition to other mechanisms like receptor stimulation, 1-aminoadamantanes may act in parkinsonian patients by enriching the transmitter content in the synaptic cleft.


Dopamine-sensitive adenylate cyclase activity in the striatum in Parkinson's disease. Dopamine (DA) sensitive adenylate cyclase activity was measured in homogenates of caudate nucleus and putamen from frozen brain of eight Parkinsonian patients and nine controls. In the control group, there was no difference in enzyme activity in respect to sex or age (21--77 years old). Caudate nucleus and putamen exhibited similar mean activity. In the Parkinsonian group, there was a significant decrease in both basal (by 50%) and DA stimulated (by 80%) activities compared with that of control group. This suggests that there may exist a functional disturbance in the postsynaptic (post-DA-nergic) region in the striatum of patients with Parkinson's disease; in addition, the decrease in basal as well as DA stimulated activity as measured in homogenates may not be the biochemical substrate for the hypothetical "denervation supersensitivity" expected to occur in Parkinson's disease.


Brain norepinephrine, dopamine, and 5-hydroxytryptamine in magnesium-deprivation encephalopathy in rats. The effect of Mg deficiency on the regional concentrations of norepinephrine, dopamine, and 5-hydroxytryptamine in the brain was studied in clinically symptomatic young rats fed a diet low in magnesium for 10 days. Decreases in magnesium concentration in the brain were not accompanied by any significant changes in these monoamines.


Distribution of PNMT and epinephrine in the medulla oblongata of normotensive and spontaneous hypertensive rats. The distribution of the Epinephrine forming enzyme (PNMT) activity and Epinephrine (E) levels was investigated in the medulla oblongata of spontaneous hypertensive rats (SH-rats) and in two normotensive strains, namely Wistar Kyoto rats (WK-rats) and Wistar rats. The PNMT activity increases progressively from the caudal to rostral parts in the C1 and C2 regions of the medulla oblongata. The enzyme activity and the E levels are in all parts of the C1 and C2 regions higher in Wistar rats than in WK-rats. The PNMT activity in all parts of the C2 region (with the exception of the caudal region), and in the middle part of the C1 region is higher in SH-rats than in WK-rats. The E levels in the SH-rats are higher than in WK-rats in the mediocaudal parts of the C2 and C1 regions.


Effects of light and of testosterone derivatives or antiandrogens on the secretory content of the subcommissural organ of the quail, Coturnix coturnix japonica. Photosensitivity of the quail subcommissural organ (SCO) is demonstrated. When the birds are submitted to continuous light the SCO cells appear depleted in secretory material and the enzyme activities investigated are of greater intensity. Such a diminished secretory content is observed in birds exposed to short daylength when they are injected with testosterone derivatives. Besides, injections of antiandrogenic compounds in continuously illuminated quails abolish the decrease of the secretory material induced by photostimulation. These results indicate that the SCO responsiveness to light appears to be mediated by some hormonal mechanism. Moreover it is more than a generalized metabolic effect: the response to antiandrogen injections suggests indeed that the SCO is a likely target organ for hormonal action.


Ultrastructure and possible function of an atrial paraganglion not directly related to atrial ganglion in the albino mouse. The ultrastructure of a paraganglion found in right atrium of albino mouse close to the superior cava vein is described. A capillary, which occupied the central part of the paraganglion, was surrounded by three chromaffin cells. Two other chromaffin cells were also present, but neither nerve fibers nor nerve endings were seen. The possible physiological role of paraganglia is discussed.


Biochemical studies of taste sensation. VII. Enhancement of taste stimulus binding to a catfish taste receptor preparation by prior exposure to the stimulus. The taste receptor membrane fraction (Fraction P2) was prepared from a homogenate of the taste tissue of the channel catfish Ictalurus punctatus. This included the rostral, dorsal, and dorsolateral surfaces of the catfish in addition to those of the barbels. The yield of Fraction P2 is 4-7 mg protein from an individual fish, with a purification averaging 8- to 15-fold over that of the crude whole homogenate and essentially quantitative recovery of binding activity in Fraction P2. Treatment of Fraction P2 in vitro with a high concentration of the taste stimulus molecule L-alanine led to a several-fold enhancement of binding activity. Enhancement of the binding of 3H L-alanine was observed after treatment with unlabeled 10 mM L-alanine and removal of the L-alanine by washing. Enhancement occurred whether the preparation was stored frozen (-65 degrees C) for an extended period in the presence of the L-alanine, or merely exposed to it in the cold without freezing. D-Alanine enhanced the binding activity of 3H L-alanine to about 60% of the level induced by L-alanine. Nonspecific binding of 3H L-alanine was unaffected by the treatment. Scatchard analyses of saturation curves for binding of 3H L-alanine to freshly prepared Fraction P2 and to L-alanine-treated Fraction P2 revealed no change in the KD value, but a several-fold increase occurred in the amount bound. Binding activity is operationally defined. Because the enhancement observed here is reminiscent of an increase in transport due to a countertransport effect, further studies were carried out to examine whether the phenomenon reflects transport or true binding. The measured binding was not increased in the presence of Na+, indicating that it is not due to an Na+-coupled transport of L-alanine. When Fraction P2 was preloaded with L-alanine (10(-6)--10(-2) M) prior to assay, no stimulation of binding was observed; instead, binding decreased. This result is consistent with a true binding phenomenon but not with a carrier-mediated transport process to explain the enhancement phenomenon. Binding assays carried out over a range of osmolarities revealed decreased binding at high osmotic strengths, suggesting that a significant portion of the ligand might be contained in vesicles. It is postulated that "hidden" or "buried" receptor sites exist in the Fraction P2 as isolated, and that these are exposed upon perturbation of the membrane structure by a high ligand concentration.


Central inhibition of an identified mechanosensory interneuron in the crayfish. Inhibitory input from crayfish mechanoreceptors is mediated polysynaptically to sensory interneurons. An identifiable sensory interneuron, the caudal photoreceptor (CPR), has been used as a model system to characterize inhibitory intermediate cells. A survey of the abdominal connectives, by antidromic stimulation, has identified eleven inhibitory cells, some of which also function as ascending sensory interneurons. These results indicate that lateral interactions within networks of mechanosensory interneurons form an integral part of the information processing mechanisms.


L9 modulation of gill withdrawal reflex habituation in Aplysia. Repeated tactile stimulation of the siphon in Aphysia normally results in habituation of the gill withdrawal reflex and a concomitant decrease in the amplitude of the excitatory synaptic input ot gill motor neurons in the abdominal ganglion. It was found, however, that induced low-level tonic activity in motor neuron L9, which does not itself elicit a gill withdrawal movement, prevented habituation of the reflex from occurring. Further, in preparations already habituated, this tonic low-level activity brought about a reversal of habituation. Although tonic L9 activity prevented the occurrence of habituation or brought about its reversal, it did not interfere with the synaptic decremental process which normally accompanies gill reflex habituation. Motor neurons L7 and LDG1 were found not to possess this ability of L9 to modulate gill reflex habituation. Evidence suggests that L9's modulatory effect is mediated in the periphery, in the gill and not centrally in the abdominal ganglion.


An endogenous motor program for sand crab uropods. A stereotyped pattern of spontaneous, rhythmic bursting in motoneurons of three principal uropod muscles in the sand carb Emerita analoga has been recorded from a deafferented chain of the four most-posterior abdominal ganglia. This endogenous motor program resembles the electromyogram pattern recorded from return-stroke and power-stroke muscles in swimming crabs in that (1) latencies of power-stroke bursts and burst periods are positively correlated with each other and (2) durations of power-stroke bursts are brief and nearly invarient. The endogenous program differs from the electromyogram pattern in having longer periods and return-stroke bursts which are brief and sporadic. The neural oscillator underlying the endogenous motor program, therefore, appears to drive the power stroke. Circumstantial evidence suggests that it may also inhibit return stroke motoneurons concurrently with excitation of the power-stroke excitor.


Dopamine beta-hydroxylase distribution in density gradients: physiological and artefactual implications. Knowledge of the vesicular origin of circulating dopamine beta-hydroxylase (DbetaH) is indispensable for any attempts to explain the parallelism or lack of it between circulating enzyme and catecholamines as they may relate to physiological stress, forms of hypertension, neurological disorders, and the response to pharmacological agents. The present study represents an effort to evaluate and to place in proper perspective data based on the DbetaH activity found in the region of the light vesicle peak of noradrenaline (NA), which is used as a quantitative measure of a population of small terminal vesicles. Distributions of vesicles and subvesicular components are compared with DbetaH and NA in sucrose-D2O density gradients used to prepare relatively pure fractions of large dense cored vesicles (LDV) from bovine splenic nerve. Although NA in sedimentable particles of the light vesicle peak is likely to be a valid measure of a small vesicle population, the following is demonstrated: (1) A substantial fraction (25%-37%) of the total sedimentable DbetaH activity can be proven to distribute in the region of the light vesicle peak from a tissue with an insignificant small vesicle population. Based on studies of vesicles from sequential nerve segments, this enzyme activity probably corresponds to a population of "immature" LDV which are undergoing axoplasmic transport and have not synthesized their full complement of transmitter. (2) Physical lysis which depletes the matrix of LDV causes redistribution of DbetaH activity from the heavy vesicle peak into the region of the light vesicle peak. Analogously, DbetaH associated with exocytosed LDV and retrograde transport particles is also likely to contaminate the region of the light vesicle peak. (3) Based on available data, it can be calculated that each small dense cored vesicle could contain only 0.1-0.5 molecules of DbetaH and that a contamination of only 0.016% LDV can account for all of the DbetaH reported to occur in the light vesicle peak of normal rat vas deferens preparations.


Catecholamine-rich cells and varicosities in bovine splenic nerve, vesicle contents and evidence for exocytosis. The bovine splenic nerve trunk contains mast cells, ganglion cells, small intensely fluorescent (SIF) cells, and varicosities which exhibit a brilliant fluorescence characteristic for noradrenaline (NA) and dopamine (DA) after formaldehyde exposure. All these catecholamine-rich structures could contribute particles to isolated nerve vesicle fractions. Mast cells are recognized ultrastructurally by their large (300-800 nm) dense granules. SIF cells may be represented by cells and processes containing dense cored vesicles (120-140 nm) which are larger than the typical vesicles in axons and terminals. Terminal-like areas with typical large dense cored vesicles (LDV, 75 nm) and small dense cored vesicles (SDV, 45-55 nm) probably correspond to the fluorescent varicosities. The LDV constitute about 40% of all vesicles in terminal-like areas and terminals. Their staining properties indicate the presence of protein, phospholipids, and ATP. Tyramine depletes NA without loss of matrix density. The LDV can fuse with the terminal membrane, and released material outside omega profiles is interpreted to depict exocytosis. Large and small vesicles are easily distinguished from the very large mast cell granules and the moderately dense Schwann cell vesicles. Neither appear to contaminate the LDV fractions but the latter may contain a small population of SIF cell vesicles. Golgi vesicles from the Schwann cells mainly occur in the lighter zones of the gradient.


Duplication of a spontaneously active neuron in Aplysia: electrical coupling and effects of a phosphodiesterase inhibitor. An abdominal ganglion from an Aplysia californica is described, in which cell R15 has anomalously duplicated. The two cells exhibited a high degree of electrical coupling, assuring functional synchrony of output in the cells, which are characterized by a complex firing pattern. Exposure of this ganglion to the phosphodiesterase inhibitor IBMX caused a more altered firing rhythm in one of the cells, as well as an enhanced inhibitory component associated with the coupling potentials between cells, resulting in a loss of synchrony between the two cells.


Binding of N-methyl isatin beta-thiosemicarbazone-copper complexes to proteins and nucleic acids. N-Methyl isatin beta-thiosemicarbazone-copper complexes interact with nucleic acids and proteins as shown by ultraviolet (UV) and visible spectroscopy and Sephadex exclusion chromatography. The Cu++ ions are most effective; Co++ ions have less albeit significant activity. Chelating agents, such as Tris and histidine, high NaCl concentration, and dimethyl sulfoxide reduce the binding of the drug-metal complex. The binding constant of the drug-copper complex to calf-thymus DNA was calculated to range between 6.9 x 10(4) and 2.7 x 10(5) M-1.


Moya-moya disease: clinical and pathological report of a case with associated myopathy. A British born white man, age 51 years, presented with cerebrovascular insuffciency, and was found radiologically to have moya-moya disease. He also suffered from a let onset type of progressive myopathy. Five months after the first signs of cerebrovascular disease he died of massive cerebellar infarction. At necropsy the brain showed multiple old and recent infarctions. The anterior cerebral, middle cerebral, and posterior communicating arteries showed segments of narrowing and occlusion by delicate connective tissue, with marked thickening and folding of the elastic lamina. Many dilated thin-walled vessels without a muscle layer and with a poorly formed elastic lamina were present in the subarachnoid space, these probably being newly formed collateral vessels. The cause of the occlusive lesions affecting the cerebral arteries is not known but the morphological appearances do not fall within any known aetiology. It seems that moya-moya disease could be regarded as a distinct entity.


Correlation of CAT scan and visual field defects in vascular lesions of the posterior visual pathways. Thirty-nine patients with various types of isolated homonymous hemianopias resulting from ischaemic lesions in the posterior parts of the cerebral hemisphere was examined by CAT scanning. Most had localised low density lesions withing the distribution of the posterior cerebral artery. The location of the lesion (deduced from a separate anatomical study of postmortem brain cut in the plane of the CAT scanner) was correlated with visual field defects. Lesions giving rise to quadrantic defects were smaller than those causing total hemianopias; lower quadrantic defects tended to occur in superior cuts and vice versa. Macular sparing was associated with survival of the occipital pole in some instances. Bilateral cases had a higher prevalence of associated defects.


Restitution of visual function in patients with cerebral blindness. Patients with postchiasmatic visual field defects were trained at the border of their visual field. Using a psychophysical method, light-difference thresholds were determined repeatedly in this visual field area. Improvement in contrast sensitivity and increase in size of the visual field could be obtained by this training procedure. The improvement was confined to the trained visual field area and showed interocular transfer indicating its central nature. Althoughh only contrast sensitivity was trained, the observed improvement was not limited to this visual function. Visual acutity, critical flicker fusion, and colour perception also showed and improvement suggesting an association of these functions. The improvement was restricted to the training period-no spontaneous recovery was observed between or after the periods of training. It is suggested that a lesion in the central visual system does not always result in a complete and permanent loss of function. The critical level of function that normally has to be reached for sufficient neuronal sensitivity may be obtained by systematic visual stimulation in the area between the intact and blind parts of the visual field. This increase in neuronal sensitivity leads to an improvement in visual performance.


Visual and somatosensory evoked cortical potentials in multiple sclerosis. The diagnostic value of the pattern reversal evoked cortical potential (VEP) and the somatosensory evoked cortical potential (SEP) has been compared in 50 patients with established or suspected multiple sclerosis. A prolonged latency of VEP was found in 96% of definite cases of multiple sclerosis, 58% of probable cases, and 20% of possible cases. A prolonged latency of SEP by stimulation of median or peroneal nerves or both was found in 86% of definite cases of multiple sclerosis, 83% of probable cases, and 50% of possibe cases. When combining the results of all three tests the diagnostic yield increased to 100%, 92%, and 50%, respectively.


Stapedius reflex in multiple sclerosis.

Stapedius reflex in multiple sclerosis. An analysis was carried out of recordings of the crossed stapedial reflex response in a series of normal subjects and 30 patients with multiple sclerosis. Parameters measured included latency, contraction time, amplitude, and relaxation time. Criteria derived from the normal group and applied to the multiple sclerosis group showed clear evidence of abnormality in 10 patients. This can be attributed to lesions in the pathways of the second and third order neurones in the trapezoid body and superior olivary complex. Present evidence suggests that further refinements in the measurement technique might significantly increase the detection rate of abnormalities in multiple sclerosis.


Prolonged azathioprine treatment of non-remitting multiple sclerosis. Two groups of 85 and 42 ambulatory patients with moderately advanced non-remitting multiple sclerosis were treated for six to 14 and three to six years with daily azathioprine. Less than 10% of these patients became confined to a wheelchair. This far exceeds any possible result in a group of non-remitting multiple sclerosis patients not so treated.


Frequency resonance investigation of the H reflex. Results of a previous study demostrated transformation of the sustained activity of a muscle into rhythmic, tremor-like activity by stimulation of different motor centres. The optimum for this transformation was in the 4-7 Hz frequency band (resonance band). Sustained rhythmic stimulation with stimuli at 0.1-20 per second caused a normal frequency curve of the H reflex in subjects with intact motor systems. The resonance phenomenon of the frequency curve of the H reflex was in agreement with that found in previous studies stimulating the different motor centres. As this requires connections between spinal and higher motor centres, it is suggested that this investigation is appropriate for a detailed functional study of spinal cord lesions.


Benign recurrent vertigo.

Benign recurrent vertigo. Patients with recurrent vertigo in the absence of cochlear signs remain a diagnostic problem. The absence of a standard system of nomenclature further hampers the understanding of these disorders. The term benign recurrent vertigo is suggested as a useful term to characterise many of these patients. The disorder shares some of the features of migraine, and seems likely to have a similar vasospastic aetiology.


Characterisation of the mobility on isoelectric focusing of individual proteins in CSF and serum by immunofixation. The mobility of 17 different proteins in CSF and serum on isoelectric focusing was investigated by subsequent immunofixation using monospecific antisera. Individual proteins yielded identical, often complex band patterns in normal CSF and serum, except transferrin which gave one to two additional bands between pH 5.8-6.4, and the low molecular wieght beta-trace protein and gamma-trace protein, which gave three bands at pH 7.4, 8.0, and 8.4, and a single band at pH 9.5, respectively, on investigation of CSF but not serum. Polyclonal IgC migrated as multiple bands between pH 4.7-8.6. Oligoclonal IgG in CSF in multiple sclerosis and neurosyphilis migrated between pH 8.6-9.5 and was easily discriminated from other proteins.


Five cases of intracranial tuberculoma followed by serial computerised tomography. Five cases of intracranial tuberculoma in Asian adults are described, all of whom presented with epilepsy. The computerised tomographic (CAT) scanner displayed the lesions particularly well, and their characteristics are described. Only two cases were explored surgically, but triple antituberculous chemotherapy has the scan abnormalities in all of them to resolve completely. It is concluded that Asian patients with lesions likely to be tuberculous should be given a trail of antituberculous drugs, and only explored if they should deteriorate clinically, or if the lesion should fail to respond on serial scans.


Tiapride in levodopa-induced involuntary movements. Tiapride, a substituted benzamide derivative closely related to metoclopramide, reduced levodopa-induced peak dose involuntary movements in 16 patients with idiopathic Parkinson's disease. However, an unacceptable increase in disability from Parkinsonism with aggravation of end-of-dose akinesia led to its cessation in 14 patients. Tiapride had no effect on levodopa-induced early morning of "off-period" segmental dystonia. These results fail to support the notion that levodopa-induced dyskinesias are caused by overstimulation of a separate group of dopamine receptors.


Parkinsonism after traumatic childbirth. Three cases of an extrapyramidal syndrome after traumatic childbirth are described.


Reduction of renal calcium reabsorption in man by consumption of dietary protein. This experiment was designed to test whether protein consumption reduces the amount of filtered calcium reabsorbed by the kidney. Nine subjects were each fed meals containing 18 g protein and 54 g protein. The intake of energy, sodium, calcium, phosphorus, magnesium and zinc was similar in the two meals. For 4 hours after the meal, measurements were made of serum calcium (total and filterable), serum creatinine, and urinary calcium, creatinine, zinc and nitrogen. Calcium reabsorption was calculated in five clearance periods, as (filterable calcium X GFR) minus urinary calcium. Urinary calcium, zinc and nitrogen were significantly higher between 2 and 4 hours after consumption of the high protein meal. Protein level did not affect urine pH or volume, serum total or filterable calcium & or GFR. The percentage reabsorption of filtered calcium was significantly lower 0.5 hours after the high protein meal, so that at 2.5 hours, reabsorption was 98.0% compared to 98.7% after the lower protein meal. We conclude that protein consumption reduces the amount of calcium reabsorbed by the kidney.


Assessment of the essentiality of methionine, threonine, tryptophan, histidine and isoleucine in immature dogs. A completely purified L-amino acid diet was used to evaluate the indispensability of methionine, threonine, tryptophan, histidine and isoleucine in the immature dog. Immature dogs fed diets devoid of methionine, threonine, tryptophan, histidine or isoleucine had severely depressed food intakes, accompanying weight loss, and a distinctly negative nitrogen balance. Partial or total removal of these amino acids resulted in a marked increase in blood and urinary urea. Estimates are made for the dietary requirements for these amino acids in the immature dog.


Toxicity of rac-1(3)-palmitoyl glycerol in weanling mice. Reinvestigation of the previously reported toxicity of saturated fat on weanling mice has shown rac-1(3)-palmitoyl glycerol to be a more potent toxic agent than free palmitic acid when fed as the sole source of dietary fat. As shown before, protection against this toxicity can be afforded by the addition of 2 to 4% safflower oil. We have now shown that if the rac-1(3)-palmitoyl glycerol is acetylated the toxicity is much less. The protective effect of diacetyl rac-1(3)-palmitoyl glycerol cannot be totally ascribed either to the presence of acetate itself or to the blocking of the free hydroxyls of palmitoyl glycerol by acylation. In vivo absorption studies coupled with in vitro experiments with pancreatic lipase suggest that the major protective effects result from increased lipolysis of the acylated palmitoyl glycerol, causing conversion to the less toxic free palmitic acid.


Lysine and arginine requirements of the domestic cat. Domestic kittens were used in four experiments to establish quantitative requirements for lysine and arginine. A purified L-amino acid diet (by calculation, 4,700 kcal metabolizable energy/kg diet) was employed throughout. Weight gain, gain:feed and nitrogen retention data of cats fed dietary lysine levels ranging from 0.48 to 1.92% suggested a requirement not exceeding 0.80%. The dietary arginine requirement for maximal gain was assayed at this level of lysine and found to be not greater than 0.83%. A dietary arginine level of 0.33% resulted in vomiting and extreme lethargy within 4 hours of ingestion.


Insulin dependent and independent actions of dietary protein on in vitro protein synthesis in skeletal muscle of rats. Insulin dependency of the effect of dietary protein on in vitro protein synthesis in skeletal muscle was studied in rats. Re-feeding fasted rats with an adequate protein diet caused an increase in protein synthetic activity of skeletal muscle within only 5 hours with a concomitant increase in concentration of polyribosomes. During re-feeding of a protein-free diet muscle protein synthesis increased significantly but it was somewhat lower than in rats fed an adequate protein diet. The concentration of insulin in plasma increased promptly and markedly when the rats were re-fed either of these diets. However, the degree of increase was slightly less in rats fed a protein-containing meal. Re-feeding normal rats with egg albumin alone, after fasting 1 day, produced a marked increase in protein synthetic activity of skeletal muscle with a parallel increase in proportion of polyribosomes in the tissue. The stimulatory action of dietary protein on muscle protein synthesis was not abolished even in streptozotocin-diabetic rats. However, there was no appreciable change in ribosomal profile in the diabetic rats. These results indicate that the function of dietary protein in regulating muscle protein synthesis is not mediated entirely by insulin.


Effect of level of protein intake on calcium metabolism and on parathyroid and renal function in the adult human male. Mechanisms involved in the hypercalciuria caused by high levels of protein intake were investigated. Six healthy males participated in a 20-day metabolic study. During the first 10-day period, all subjects were given a 47 g protein diet and during the second 10-day period, a 142 g protein diet. Calcium, magnesium and phosphorus intakes were kept constant at 515, 320 and 1,110 mg daily, respectively. Urinary calcium was elevated significantly when the protein intake was increased. Glomerular filtration rate and calcium clearance were increased significantly when the high protein diet was fed; the fractional tubular reabsorption of calcium was decreased from 98.4 to 97.4%. Thus, the increase in urinary calcium caused by the high protein diet appears to be due in part to an increase in the filtered load of calcium by the glomeruli and in part to a decrease in calcium reabsorption by the renal tubules. The level of protein intake had no effect on the fasting serum concentrations of parathyroid hormone, total calcium, magnesium or inorganic phosphorus or plasma ultrafiltrable calcium.