System for simplified discontinuous-gradient polyacrylamide-gel electrophoresis. We describe a modified polyacrylamide gel electrophoresis method with which plasma proteins are excellently resolved in 4 h. We made a three-layer discontinuous gradient gel with concentrations of 80,60, and 45 g of polyacrylamide per liter of tris(hydroxymethyl)aminomethane/ethylenediaminetetraacetic acid/borate buffer, pH 9.2. Time and material needed in the preparation of the gel was greatly decreased, a well-resolved (22-25 bands) pattern was obtained in 1 h, and the gel slabs were easier to remove from the cell than were gels prepared according to the manufacturer's specifications. We saw a marked shift in the higher polymers of haptoglobin type 2-2 toward the cathode in this system, and also a clearly resolved double band of alpha2-macroglobulin in whichthe more anodal band appears to represent alpha2-macroglobulin/protease complex.


Non-fluorescent Y chromosome in a male infant with Turner's symptoms and XO/XY mosaicism. A 45, X/46,XY mosaicism was found in a male infant with stigmata of Turner's syndrome but normal male external genitalia. In contrast to the Y chromosome of his father, the Y chromosome of the patient does not display either the characteristic brilliant fluorescence or the typical dark heterochromatin staining of the distal long arm. Furthermore, DNA replication in the abnormal Y chromosome was shown to be premature. Mechanisms leading to the observed abnormalities are discussed.


Hydration of arene and alkene oxides by epoxide hydrase in human liver microsomes. The comparative hydration of styrene 7,8-oxide, octene 1,2-oxide, naphthalene 1,2-oxide, phenanthrene 9,10-oxide, benzo[a]anthracene 5,6-oxide, 3-methylcholanthrene 11,12-oxide, dibenzo[a,h]anthracene 5,6-oxide, and benzo[a, 7,8-, 9,10-, and 11,12-oxides to their respective dihydrodiols was investigated in microsomes from nine human autopsy livers. The substrate specificity of the epoxide hydrase in human liver microsomes was very similar to that of the epoxide hydrase in rat liver microsomes. Phenanthrene 9,10-oxide was the best substrate for the human and rat epoxide hydrases and dibenzo[a,h]anthracene 5,6-oxide and benzo[a-a)pyrene 11, 12-oxide were the poorest substrates. Plotting epoxide hydrase activity obtained with one substrate against epoxide hydrase activity for another substrate for each of the nine human livers revealed excellent correlations for all combinations of the 11 substrates studied (r = 0.87 to 0.99). The data suggest the presence in human liver of a single epoxide hydrase with broad substrate specificity. However, the results do not exclude the possible presence in human liver of several epoxide hydrases that are under similar regulatory control. These results suggest the need for further investigation to determine whether there is a safe epoxide of a drug whose in vivo metabolism is predictive of the capacity of different individuals to metabolize a wide variety of epoxides of drugs and environmental chemicals.


Herpes zoster: a combined therapy regimen. A combined method of therapy for the routine management of herpes zoster in most all age groups is presented. The uniformly gratifying results of this treatment prompted this report. A total of 105 known cases of acute herpes zoster in which this regimen was employed during the past 15 years was analyzed in this study. Clinical observations before and after treatment are presented with discussion of clinical factors, treatment and conclusions.


Ionophores. Chemistry, physiology and potential applications to bone biology. Ionophores are substances which facilitate the movement of ions into and/or through organic phases. The two major classes of compounds are the neutral ionophores, which form charged clathrate complexes with ions, and the carboxylic polyether ionophores which are neutral in the complexed state. Many of the ionophores exclusively bind organic monovalent cations. Selectivity for divalent metal cations or for anions is less common. One carboxylic polyether, X-537A, forms complexes with small organic cations as well as a variety of metals. Ionophores selective for monovalent cations have been used primarily for studies of mitochondrial metabolism. Compounds like X-537A and A23187, which show high affinity for biologically active divalent cations, are being widely used as tools to study the physiologic roles of these cations. These studies have confirmed the central role of calcium in many biological processes. The major problem encountered with X537A is a lack of selectivity. A23187 has produced variable results in studies of bone as well as other tissues. Despite these difficulties, the ionophores offer promise of being useful tools in studies of bone metabolism.


Radioactive isotope labeled diffusible components of a bone morphogenetic substratum. Bone matrix gelatin labeled with radioactive glucose, cysteine, proline, tyrosine or tryptophan was enclosed in one chamber while an adjoined second chamber was left empty to become filled with interstitial fluid. The paired diffusion chambers were implanted into the anterior abdominal wall of allogeneic rats for 14 days. Release of the matrix gelatin (BMG) from the loaded chamber was followed by a bone formation on the outer membrane of the unloaded chamber. 3H-cysteine labeled components of BMG were disseminated into the interstitial fluids and throughout the host tissues 5 times more rapidly than 3H-proline labeled BMG. The relative rates of loss of radioactivity from the residue was cysteine greater than tryptophan greater than tyrosine greater than proline. Electron microscopic autoradiography revealed random dissemination of collagenous and non-collagenous derivatives of BMG. 3H-glucose distribution was coordinated with ruthenium red staining coarse granules and may have originated from BMG. A component of interstitial fluid staining positively with Coomassie brilliant blue and PAS methods was identified by SDS polyacrylamide gel electrophoresis of interstitial fluid aspirated from empty chambers attached to chambers loaded with bone matrix gelatin. The character of the bone morphogenetic property in bone gelatin has not yet been defined in physico-chemical terms but present evidence points to the view that it is associated with a protein molecule.


Herpes zoster in the elderly.

Herpes zoster in the elderly. Herpes zoster is a self-limited disorder which in most cases resolves without complications. The specific defect in host immunity that permits activation of latent V-Z virus and the occurrence of herpes zoster in both healthy and debilitated individuals has not yet been identified. In some patients, particularly the aged, complications occur during the acute phase of the disease or there are sequelae that may incapacitate the patient later. The most important of these is postherpetic neuralgia. In the elderly the chance of developing neuralgia following herpes zoster is about 50%. Involvement of the eye may produce minimal scarring or permanent blindness. There is an increasing incidence and severity of herpes zoster in association with malignant disease and in particular with Hodgkin's disease. Treatment of herpes zoster in the elderly should be determined by presenting symptoms. Topical medication such as the basic shake lotion is helpful. Personal experience and published reports suggest that early systemic administration of corticosteroids to healthy patients with severe herpes zoster pain with lessen the occurrence of postherpetic neuralgia. Administration of herpes zoster immune globulin is only effective in reducing the morbidity or preventing varicella in high risk individuals. ZIG does not affect the clinical course of herpes zoster.


Localisation of non-replicating heterochromatin in polytene cells of Drosophila nasuta by fluorescence microscopy. A simple fluorescence technique is decribed to localise in situ the non-replicating alpha heterochromatin in the chromocentre region of Drosophila nasuta polytene nuclei. After incorporating 5-bromodeoxyuridine in larval salivary gland cells for one or two cycles of replication, the polytene nuclei are examined for Hoechst 33258 flourescence at pH 7.0. The non-replicating alpha heterochromatin remains brightly fluorescing as it does not incorporate any 5-bromodeoxyuridine while the rest of the replicating chromatin shows dull fluorescence due to the quenching of Hoechst 33258 fluorescence by the bromodeoxyuridine substituted DNA.


[Bleomycin lung (author's transl)].

[Bleomycin lung (author's transl)]. Histological and electron-microscopic study of the lungs of 15 patients who had been treated with bleomycin for advanced squamous cell carcinoma demonstrated marked histological changes in nine. They were typical of bleomycin effects: alveolitis, intra-alveolar and interstitial oedema, pulmonary hyaline membranes, disseminated intravascular coagulation, intraalveolar and interstitial fibrosis, atelectasis, metaplasia and dysplasia of the alveolar lining cells. These lesions had a focal distribution, preferentially in the subpleural and periseptal regions. Each of these lesions alone is a non-characteristic reaction, but their combination makes it a distinct entity (bleomycin lung). Three different clinical courses were noted: (1) cases with no or little abnormality; (2) acute form during or shortly after bleomycin treatment; (3) chronic, progressive form of bleomycin lung which may end fatally as late as 1 1/2 years after bleomycin treatment had been discontinued. Squamous cell metaplasia is the most characteristic sign of bleomycin lung. It should not be confused with pulmonary metastases. To prove the diagnosis of bleomycin lung often requires systematic histological investigation. A schema of the pathogenesis of the bleomycin lung is proposed in which the formation of microthrombi plays an important part.


Types of hypoxic and posthypoxic delta activity in animals and man. 1. Two types of hypoxic delta activity were observed in ECoG records of dogs subjected to exsanguination, sudden cardiac arrest resulting from ventricular fibrillation or mechanical asphyxiation, as well as during post-hypoxic recovery. They were polymorphous delta activity (PDA) and "standard slow complexes" (SSCs). 2. These two types of delta activity were found to differ from each other as to the shape of the potentials; their amplitude in monopolar and bipolar leads; the cortical areas in which the activity exhibited the highest amplitude; the degree of manifestation in the cortex and subcortical structures; their relation to activating influences; their dependence on the stage and character of hypoxia. 3. Analysis of EEG records of patients during and after hypoxia made it possible to distinguish both PDA and a type of delta activity similar to the SSCs observed in experimental animals. 4. It is suggested that the appearance of SSCs is determined by the activity of a pacemaker situated in the diencephalon and responding to pO2 changes, and that the mechanism of SSC generation is closer to that of monorhythmic, bilataerally synchronous delta waves observed in primary irritative or epileptogenic processes in the diencephalon.


Behavioral and hippocampal electrical changes during operant learning in cats and effects of stimulating two hypothalamic--hippocampal systems. The electrical activity of the dorsal and ventral hippocampus and of the anterior and posterior neocortex is described and illustrated during the development of trained operant behavior in water-deprived cats learning to bar-press for water reward under three operant conditioning schedules: (a) continuous reinforcement; (b) alternating 10 sec periods of reinforcement and non-reinforcement; and (c) mixed reinforcement and non-reinforcement with special auditory and visual cues. In addition, the contrasting effects of 100 c/sec electrical stimulation of the medial and lateral hypothalamic systems upon hippocampal electrical activity and operantly trained behavior are reported. During the early stages of learning to bar-press for water reward, when close attention to the bar and water-well are required, hippocampal electrical activity manifests synchrony (theta rhythm) and neocortical electrical activity is desynchronized. Subsequently, as bar-pressing performance improves and requires little attention to the manipulanda, the pattern of hippocampal and neocortical electrical activity is one of irregular slow waves mixed with low voltage high frequency activity, characteristic of relative inattention and automatic performance. During alternating or mixed reinforcement and non-reinforcement schedules of operant training differential effects are observed during reinforcement and non-reinforcement periods, the latter being characterized generally by lower voltage, mixed low and high frequency activity, except when orienting and shifting of attention occurred with associated theta rhythm bursts. Stimulation of the medial hypothalamic system has a striking inhibitory effect upon bar-pressing for water reward. Bar-pressing ceases for many minutes but its eventually resumed at the pre-stimulation rate. In contrast, stimulation of the lateral hypothalamic system only interrupts bar-pressing for a matter of seconds. Possible causes of these differential effects, and especially the prolonged inhibition of bar-pressing induced by medial hypothalamic stimulation, are discussed.


Ocular accomodative changes in humans induced by positional changes with respect to gravity. Ocular accomodation was measured in human subjects while they were rotated at 1 degree/sec about their naso-occipital axes. Sixteen normal subjects were tested with 45 complete and 27 partial revolutions. Naso-occipital rotations ipsilateral to the eye being observed caused accommodative, lens-thickening changes. This effect begin at about 14 degrees from head upright position, tended to reach a maximum by 45 degrees and usually stayed at this level until about 90 degrees. The values tended to return to control level by 180 degrees. With naso-occipital roll in the direction opposite to the eye being observed, there was little change until about 135 degrees at which point further roll was typically clear accomodation. This continued to a maximum at about 270 degrees rotation, and at about 350 degrees returned to control values. Arguments are presented relating this response to the utricles, which are approximately parallel to earth horizontal. The threshold of this response, between 3.7 and 77 cm/sec2, is similar to the threshold of linear acceleration when measured by other means.


A cortical evoked potential that reflects the conditioned, positive incentive value of the stimulus. I. The evoked potential. As squirrel monkeys learned that a tone cue signalled the availability of a food reward there appeared in pre- and post-central cortex a surface negative waveform in response to the cue. This waveform disappeared, concomitantly with the animal's behavioral response, during extinction and pseudoconditioning. It was independent of changes in cue intensity sufficient to markedly alter earlier components of evoked activity and it disappeared when evoked activity was averaged with the behavioral response, rather than the cue, as reference point. It was concluded that this evoked potential reflects cortical activity somewhere in the stimulus-response sequence between the recognition of the cue and the decision to act.


A cortical evoked potential that reflects the conditioned, positive incentive value of the stimulus. II. Drive motivation (hunger). It was shown that the late negative wave evoked in squirrel monkey cortex by tone cues signalling the availability of reward was smaller when the animal did not respond to the cue and often disappeared if the animal ignored many cues in succession. It decreased in amplitude as a test session progressed, and the rate of this decrease increased if the rate of reinforcement was increased. In some animals the size of the evoked wave was smaller if the animal was not fasted before a test session. In some animals the average size of a significant positive correlation with the number of cues responded to and a negative correlation with the animal's average reaction time. It is concluded that the evoked wave is probably a reflection of the conditioned incentive value of the cue.


A cortical evoked potential that reflects the conditioned, positive incentive value of the stimulus. III. Manipulative motivation and the value of the reward. Changing animals from a schedule on which they had to bar-press after a tone cue to get a food pellet, to a schedule on which the food pellet was presented automatically after the tone cue and the manipulandum was not available, had no effect on the latency, amplitude, or shape of the M-wave evoked by the tone cue. The amplitude of the evoked wave did, however, decrease more rapidly, as a test session progressed, with the latter schedule than with the former schedule. Failure to fast animals before test sessions markedly reduced the amplitude of the M-wave. Variation in the type of food pellet used caused marked variations in the amplitude of the evoked response. In all cases, there was a high degree of correlation between M-wave amplitude and a behavioral indication of the animal's interest in the reward. It was concluded that the M-wave reflects the conditioned incentive value of the cue, or in other words, the level of interest of the animal in the reward.


Absence of thermoregulatory vasomotor responses during fast wave sleep in cats. The skin temperature of the pinna was recorded in unrestrained cats sleeping at different ambient temperatures. The results show that during fast wave sleep vasomotion is inconsistent with homeothermic regulation as ear skin temperature increases and decreases at low and high ambient temperatures, respectively. The effects of preoptic heating during slow wave and fast wave sleep reveal that a depression of the responsiveness of hypothalamic thermoceptive structures underlies the absence of thermoregulatory vasomotor responses during fast wave sleep.


Ethanol and marihuana effects on event-related potentials in a memory retrieval paradigm. Twelve men performed the Sternberg memory retrieval task under three experimental conditions: after oral doses of marihuana extract calibrated to contain 0.7 mg/kg delta9-tetrahydrocannabinol (THC), 1.0 ml/kg 95% ethanol, or placebo. Simultaneously, the EEG was recorded from Ca to linked ears and the EOG from leads above and below the right eye. In this task, subjects saw a set of 1 to 4 digits follwed by a warning tone that was followed 1.5 sec later by a test digit. Subjects indicated by pressing one of two buttons whether the test digit was in-set or out-of-the-set. There were no drug effects on N1 in the evoked potential to the warning tone, but P3 amplitude was smaller under THC and ethanol than under placebo. CNV amplitude in the interval between the warning tone and the test digit showed no drug effects, indicating that the subject was equally prepared for the test digit regardless of drug received. However, the latency of 50% resolution of the CNV was longer under THC than under placebo. THC also increased the reaction time for each set size by about 75 msec above the values for ethanol and placebo, the latter two not differing significantly. Set size affected N1 and P3 amplitudes and latencies and CNV amplitude, as well as 50% CNV resolution latency and reaction time, but there were no drug chi set size interactions.


Hippocampal RSA (theta), apnea, bradycardia and effects of atropine during underwater swimming in the rat. Hippocampal RSA (theta), neocortical EEG, and heart rate were recorded during surface and underwater swimming in the rat. RSA was present with slightly reduced frequency during the apnea, bradycardia, and vibrissae arrest associated with under water swimming. Atropine sulfate (50 mg/kg) blocked bradycardia did not affect RSA. Contrary to previous suggestions, no causal relation was found between RSA and respiration, heart rate, or vibrissae movement. The study supports the view that RSA is related to the neural control of voluntary movement. It is suggested that the slight reduction in RSA frequency during underwater swimming may be part of an oxygen conservation process.


Theta bursts: an EEG pattern in normal subjects practising the transcendental meditation technique. In a survey of the EEG characteristics of persons practising the Transcendental Meditation technique, 21 of 78 people demonstrated intermittent prominent bursts of frontally dominant theta activity. On the average across subjects, the theta bursts occurred about every 2 min, had an average duration of 1.8 sec, and an average maximal amplitude of 135 muV. Typically, the bursts were preceded and followed by alpha rhythm. Subject reports elicited during theta bursts indicated pleasant states with intact situational orientation and no subjective experiences related to sleep. Fifty-four non-meditating controls showed no theta bursts during relaxation and sleep onset. It is hypothesized that theta burst may be the manifestation of a state adjustment mechanism which comes into play during prolonged low-arousal states, and which may be related to EEG patterns of relaxation in certain behavioural conditions.


Applications of statistical equivalence to newborn EEG recordings. Epochs were selected from sleep EEGs of newborns during both quiet and REM phases. A first order parameter analysis was done in order to test the homogeneity (run test and one way variance analysis) of the data within the epochs, follwed by a study of spectral equivalence between epochs. Comparisons were made between average spectra in the frequency range 0.5--4 c/sec; and between their band-by-band components, to test the feasibility of automatic classification of sleep stages by means of frequency analysis. On the basis of the statistical parameters we used, spectral analysis appears to be of limited value for the purpose of automatic classification of newborn EEG records; due to a lack of complete homogeneity of the spectra within each sleep state, it is not possible to identify a characteristic spectrum for active sleep and quiet sleep.


Drug effects on EEG frequency spectra as a function of interstimulus interval. The frequency spectra of EEG background activity preceding the stimulus in a serial simple RT task were investigated as a function of interstimulus interval (ISI). The percentage of power in the alpha and theta bands increased with longer ISIs while the delta activity decreased. Additionally, the effects of an amphetamine and a barbiturate were investigated. The increase in alpha activity with longer ISI conditions were larger after barbiturate than after placebo, while it remained constant after amphetamine treatment. Contrary to expectation, no systemic changes in alpha activity were found during the 3-h session for any of the treatments. It is suggested that the background EEG during the foreperiod of an RT task mainly reflects phasic arousal. The characteristics of the background EEG in the present experimental situation may be quite different from that recorded under "relaxed" no-task conditions.


Pulmonary toxicity of bleomycin.

Pulmonary toxicity of bleomycin. Diffuse pulmonary fibrosis is associated with bleomycin administration to humans. The sequential reactions of lung cells to this drug have now been investigated in mice following injection of 20 mg/kg bleomycin twice per week for 4 to 8 weeks. Cytoplasmic and subendothelial edema was first observed in large vessels and by 4 weeks involved the capillaries. The reaction in many animals did not progress further than endothelial lesions with accumulation of interstitial edema. However, 30% of mice subsequently showed necrosis of type 1 epithelium with a fibrinous exudate in the alveoli. Fibroblastic organization of the fibrin resulted in the deposition of intraalveolar collagen as well as extensive septal fibrosis by 8 weeks. Epithelial repair, normally accomplished by type 2 cell proliferation and transformation to type 1 cells, is characterized in this case by division and metaplasia of type 2 cells. The metaplastic cells were, however, capable of DNA synthesis and probably of further cell division. The results indicate that the pulmonary endothelium is the initial site of injury. Extensive damage to these cells could allow the drug access to interstitial and epithelial cells. Focal necrosis of type 1 epithelium is the critical event that triggers the exudation of fibrin and the subsequent reparative processes.


Organization of sequences of avian globin mRNA. Formamide polyacrylamide gel electrophoresis shows that chicken globin mRNA contains about 6.50 nucleotides, and since only 435 of these code for globin, a further 215 are not translated, and their function and position are not known. This work has produced the following conclusions. 1. 45-50 of these untranslated nucleotides are present as poly (A) at the 3' terminus. 2. The 3' untranslated region of chicken globin mRNA is at least 90 nucleotides in length. This minimal estimate is based on data derived from hybridization of defined lenghts of chicken globin cDNA to rabbit globin mRNA. The percentage of avian globin cDNA sequences which hybridize to rabbit globin mRNA is directly proportional to the length of the cDNA in each case. This relationship holds for lengths of cDNA from 115 up to 620 nucleotides. The low percentage homology for short cDNA molecules is not due to their being short per se. In homologous mRNA excess hybridizations (chicken cDNA/chicken mRNA), all cDNA preparations were completely protected from S1 nuclease digestion. 3. It is probable that there is greater evolutionary divergence in the 3' untranslated region of chicken and rabbit globin mRNA when compared with the coding regions of these molecules; The combined data is sued to formulate a regional map of chicken globin mRNA,


Intravascular persistence of hydroxyethyl starch in man. In two groups, each consisting of five healthy volunteers, 7 ml blood/kg body weight were exchanged with equal amount of hydroxyethyl starch (HES) and dextran 60 solutions, respectively. Dextran 60 plasma levels, determined by the anthrone method, were undetectable after 4 weeks. The elimination of HES from the blood, determined by an immunological technique and by the anthrone method, had a very protracted course. Two weeks after infusion the HES plasma concentrations were 9% of the initial value and after 17 weeks they were still above the 1% level. The prolonged intravascular persistence of HES in its commercially available preparation, and the possibility of tissue accumulation after repeated HES infusions were considered undesirable. The hypothesis that HES infusion causes and augmentation of serum alpha-amylase concentrations in man was confirmed. This effect should be borne in mind when HES solutions are given to patients in whom the diagnosis of acute pancreatitis might be considered.


Method of presentation of meaningful prose to young and old adults. Thirty young (x = 27.5 years) and thirty old (x = 67.7 years) volunteers read a selection of prose and then were tested for retention with a fifteen item multiple-choice comprehension test. Half the subjects in each group were in the standard visual condition and read the passage silently while the rest read the material aloud to obtain a simultaneous auditory-visual input. There were no age-related differences in the time taken to read the materials and, for both age groups, reading aloud required significantly more time than silent reading. Although the young subjects had significantly higher scores than old on the comprehension test, neither age group exhibited any differences attributable to mode of presentation of materials.


Metabolism of antithrombin III (heparin cofactor) in man: effects of venous thrombosis and of heparin administration. The metabolism of human antithrombin III (heparin cofactor) was studied in four control subjects, in four subjects with peripheral obliterative arterial disease, in six patients with recent venous thrombosis and in one patient with clinically severe haemophilia A. The labelled antithrombin III has a high specific activity (5.75 units/mg) and displayed a single band on SDS-polyacrylamide gel electrophoresis. On Sephadex G-100 gel filtration the labelled material eluted in the same position as the antithrombin III activity in plasma. Crossed immunoelectrophoresis of a mixture of fresh plasma and labelled antithrombin III against a specific antiserum, revealed a single precipitin line in which radioactivity was concentrated. The changes in electrophoretic mobility of both the plasma antithrombin III and the labelled material following the addition of heparin to the mixture or following coagulation were identical. The purified antithrombin III behaved as a homogeneous protein in the turnover experiments. The plasma radioactivity data were approximated by a sum of two exponential terms and the metabolism of antithrombin III represented by a two compartment mammillary model. Results in the control subjects were as follows: plasma antithrombin III concentration 19.6 +/- 2.3 mg/100 ml; intravascular fraction 0.45 +/- 0.05; fractional catabolic rate 0.55 +/- 0.02 of the plasma pool per day; half-life of the plasma radioactivity 2.83 +/- 0.26 days. Circulating large molecular weight degradation products of labelled antithrombin III could not be detected by Sephadex G-100 gel filtration. No significant differences in these parameters were found in the patients with peripheral arterial insufficiency. The turnover rate of antithrombin III was normal in the patient with haemophilia A. In three patients with venous thrombosis not treated with heparin, the turnover of labelled antithrombin III was in the normal range. In three patients with venous thrombosis, treated with heparin, the plasma radioactivity half-life was significantly shortened (2.13 +/- 0.08 days) and the fractional catabolic rate increased (0.75 +/- 0.05) of the plasma pool per day). In one of these patients, the labelled antithrombin III had been incubated with an equimolar amount of heparin prior to injection. In this patient the plasma radioactivity half-life was in the same range as in the other two patients (2.15 days).


Functional organization of the vestibular input to the anterior and posterior cerebellar vermis of cat. 1. Responses evoked by electrical stimulation (auditory division of the VIIIth nerve sectioned chronically) and natural stimulation of the vestibular apparatus were recorded in the anterior and posterior cerebellar vermis of cats anesthetized with Ketamine or Nembutal. Under Ketamine the functional state of the cerebellar cortex was similar to that of the decerebrate or encéphale isolé preparation. 2. Vestibular-evoked responses were found bilaterally throughout the vermis (lob. I-X) and parts of pars intermedia and were, for the most part, mediated via the mossy fiber-granule cell pathway although natural stimulation occasionally evoked climbing fiber responses in Purkinje cells. 3. Lesion and stimulation experiments suggested that the polysynaptic potentials recorded in the dorsal folia of the anterior and parts of posterior vermis were mediated, at least in part, by the lateral reticular nucleus. Potentials recorded in the deeper folia often had shorter latencies and were probably mediated by primary and/or secondary vestibular fibers. Studies with horseradish peroxidase (injections in lob. V and VI) supported these notions. 4. An analysis of Purkinje cell responses to sinusoidal rotation and steps of angular acceleration or velocity indicated that P-cells in these regions signalled angular head velocity in the mid-frequency range. Single canal responses as well as multi-canal convergent P-cell responses were found. Purkinje cells also responded to static head displacement.


Towards a chemical definition of idiotypy. The idiotypic determinants have been precisely located to the variable regions of immunoglobulin polypeptide chains. Both chains are generally required to express the idiotype. The major idiotypic determinants are the result of the amino acid sequence of the hypervaiable regions, although some idiotypic determinants reside outside the antibody combining site and these so called "framework idiotypes" are important markers. In my view the hypervariable regions are spatially disposed so as to present adequate antigenic stimulation, and they display enough structural heterogeneity to account for the uniqueness of the idiotype in the general population of immunoglobulin molecules. Hypervariable regions, the antibody combining site, and the idiotypic determinants thus amalgamate three formerly diverse concepts into a unified theoretical construct.


The use of idiotypes as markers for antibody variable regions in the rabbit. This article reviews the use of idiotypes as variable region genetic markers in the rabbit. Topics discussed include reagents and assays for the detection of idiotypes, evidence concerning the association of idiotypes with specific antigen binding sites on antibodies, and the inheritance of idiotypes in the rabbit. Several points are emphasized in this review. First, interpretation of idiotypic phenomenons are strongly dependent on the reagents and assays employed. Second, while strong evidence exists that a given idiotype is a marker for a specific antigen binding site, exceptions to this association have been reported. Third, the inheritance of identical or similar idiotypes has been demonstrated in several instances, but it is not always demonstrable, perhaps because an idiotype is a complex phenotype. Several reasons for this complexity are pointed out. Fourth, idiotypes are linked to group a allotypes and VL subgroups but exceptions to these associations have been described for antibodies isolated from single individuals. The significance of these exceptions is discussed. Current areas of interest in rabbit idiotypy include the relationship of idiotypes to other V region markers, the genetics of idiotypes specific to H or L chains, and the relationships among the idiotypes of antibodies isolated from a single rabbit.


Complementary idiotypy in the regulation of the immune response. A new concept is presented for the interactions of two complementary antibodies in the immune response. These antibodies bind to each other by means of their variable sequence determinants and therefore are designated as complementary idiotypes. Under certain conditions, both complementary idiotypes are produced by the same animal at the same time. An idiotype can drastically affect the expression of the complementary idiotype in the animal, inducing a peripheral quench effect of antibody-binding activity and a central effect on the immunocompetent cell, which produces the complementary idiotype. It is proposed that complementary idiotypes might be induced during every immune response, thus playing an essential role in the regulation of the immune response.


A schematic approach to the work-up of amenorrhea. Amenorrhea is a symptom having many possible causes. Since amenorrhea can result from disturbed function anywhere in the hypothalamic-pituitary-ovarian-uterine axis, a specific etiologic diagnosis must be made if treatment is to be effective. For this purpose, a diagnostic scheme for the differential diagnosis of the etiology of primary and secondary amenorrhea  is proposed. This scheme includes a progestin test, a cyclic estrogen and progestin test, a luteinizing hormone-releasing hormone (LH-RH) loading test, and a gonadotropin (human menopausal gonadotropin and human chorionic gonadotropin) loading test. A specific pattern of responses to LH-RH and gonadotropins exists in patients with hypothalamic, pituitary, and ovarian amenorrheas, respectively, and the character of the response may facilitate the etiologic diagnosis of amenorrhea. The clinical usefulness and/or value of the scheme in the diagnosis and treatment of amenorrheas is discussed.


[A case report of Hashimoto's disease with anti-thyroxine autoantibody (author's transl)]. In one case of untreated Hashimoto's disease, serum thyroxine (T4) value by radioimmunoassay (RIA) was significantly lower than that by competitive protein binding analysis (CPBA). The discrepancy was found to be due to the presence of antithyroxine autoantibody in the serum. This phenomenon was considered to be of practical importance in interpreting the T4 value by RIA in cases with autoimmune thyroid diseases. The patient was 59-year-old woman with a 30-year history of goiter. A diagnosis of Hashimoto's thyroiditis had been established by open biopsy of the thyroid ten years ago. The patient was judged to be euthyroid on the basis of clinical and laboratory evaluation (mean serum T4 by CPBA (Tetrasorb and Tetratab kit), 5.0 mug/100 ml; serum T3, 165 ng/100 ml; T3 resin uptake, 31.8%; and serum TSH, 2.0 muU/ml). TBG binding capacity was 24 mug/100 ml. Anti-thyroglobulin antibodies (anti-Tg), once positive ten years before, was negative at this time. But the mean T4 in the serum measured by T4 RIA and RIA-Mat T4 kit were 1.7 and 2.9 mug/100 ml, respectively. Recovery of the T4 added to the patient's serum evaluated by RIA-Mat T4 kit, was 71.2%, although the recovery using a control serum was 108%. Binding of 125I-T4 to the serum or fractions of the serum was studied by using polyethylene glycol (PEG) method, column chromatography, and double antibody precipitation. The results were as follows: 1) The binding of 125I-T4 to the patient's serum was detected by using RIA kit system without addition of anti-T4 serum. 2) On Sephadex G-200 chromatography of 125I-T4 incubated with the serum or the rabbit anti-T4 antibody in the presence of ANS, an early radioactive peak was observed by using the patient's serum as in the case of the anti-T4 antibody. When the serum after thermal inactivation of TBG, was incubated with 125I-T4, and was applied to the Sephadex G-200 column, a radioactive peak was observed in the area where 7S fraction was detected by protein peak. 3) The binding of 125I-T4 to the patient's IgG was 9.0% by using double antibody method when the binding to a control IgG was 0.5%. 4) The binding of 125I-T4 to IgG fractions was also proved by PEG method. 5) The binding of 125I-T4 was competitively inhibited by the addition of unlabeled T4. The affinity constant was 1.9 X 10(8) L/mol and its binding capacity was 0.8 mug/100 ml serum. From these data this T4 binding IgG was considered to be anti-T4 autoantibody. The cross reaction with T3 was approximately 8.3%. MIT and DIT did not displace labeled T4 when tested in amounts varying from 0.1 to 100 ng/assay. By using the paper electrophoresis, the binding of 125IT4 to the serum or IgG was not detectable. Therefore this method was considered unsuitable for detecting such anti-T4 antibody. As we couldn't find any significant binding of 125I-T4 to sera in 37 other patients with Hashimoto's disease by using the PEG method, the incidence of this phenomenon was considered to be low...


Relation between hypotensive effect in cats and reactions in children using normal human immunoglobulin. The paper deals with the rare adverse reactions following administration of normal human immunoglobulins in Bulgaria. Experimental work has shown that such lots cause hypotensive effect in cats and lead to the release of pharmacologically active substances, measured biologically and spectrofluorometrically. The same immunoglobulins give two peaks of IgG aggregates by gel-chromatography and possess high anti-complementary activity. The possible relation between hypotensive effect in cats and reactions in children is discussed.


Mean alpha-fetoprotein values of 1,333 males over 15 years by age groups. Sera of 1,333 African, Caucasian, Chinese and Malay males over 15 years from four countries were randomly assayed for alpha-fetoprotein (AFP) by the radioimmunoassay. The average AFP level was 2.97 IU/ml (+ 2 SD = 8.78, -2 SD = 1.00) which can be compared with previously published figures ranging from 2.3 to 1.2 ng/ml. Average values increased with age and the upper limit (mean + 2 SD) in the over 64 years age group was close to 17 IU/ml. The use of international units should facilitate the comparison of results between laboratories. The present data can help in determining whether an AFP value is "normal" and when AFP levels of certain follow-up patients revert to normal.


Detection of oncornavirus-like particles in normal and tumorigenic rat liver cell cultures. Six epithelial cell cultures established from adult Wistar rat liver were examined by electron microscopy for oncornavirus-like particles. Two heteroploid and tumorigenic cultures demonstrated many cells with virus-like particles of type-A and type-C morphology. The remaining four, composed of diploid nontumorigenic cells, demonstrated similar virus-like particles but in few cells and only after bromodeoxyuridine treatment. The virus-like particles in the tumorigenic cultures were identified as a rat type-C virus by serological assays.


Lymphocyte cytotoxicity in alcoholic hepatitis. Studies were undertaken to evaluate the cytotoxicity of peripheral lymphocytes obtained from patients with alcoholic hepatitis. Lymphocyte cytotoxicity to Chang liver cells was investigated by a microcytotoxicity test, and that to autologous liver cells obtained by percutaneous liver biopsy was studied using a 51Cr release assay. Lymphocytes from patients with alcoholic hepatitis were found to be highly cytotoxic to Chang liver cells and autologous liver cells when compared to those of healthy subjects (P is less than 0.001). Cell-free supernatant fluid of lymphocytes from patients with alcoholic hepatitis incubated with purified alcoholic hyalin for 5 days was significantly cytotoxic to Chang liver cells (P is less than 0.01), indicating that a cytotoxic factor is elaborated by sensitized lymphocytes. A significant reduction in cytotoxicity was noted with disappearance of clinical features or direct addition of a purified isolate of alcoholic hyalin or its preincubation with lymphocytes. Preincubation of sensitized lymphocytes with acetaldehyde increased cytotoxicity for autologous liver beyond that obtained by the combined effects of lymphocytes alone and acetaldehyde alone (P is less than 0.001), interpreted as evidence that ethanol toxicity and hyperactivity of lymphocytes independently and collectively contribute to development of cirrhosis in patients with alcoholic hepatitis who continue to imbibe alcohol.


Thyrotoxicosis and hypercalcaemia: response to antithyroid drugs and salmon calcitonin. Four patients with thyrotoxicosis, hypercalcaemia and metabolic bone disease are described. One of them had a 'hot nodule', T3 toxicosis and a parathyroid tumour and another had thin bones, subperiosteal cortical bone erosions and complete dysphagia. Hypercalcaemia persisted during treatment with antithyroid drugs in two patients, both of whom had hyperparathyroidism. The administration of salmon calcitonin to these two patients before starting antithyroid treatment produced an immediate and sustained fall in serum calcium and urinary hydroxyproline levels. Calcitonin administration should be of value in the early management of hypercalcaemic patients.


Oligopeptides as models for Tyr/Glu sequences in the multichain co-polymer poly(Lys)  poly(Tyr, Glu)-poly(DL-Ala). The syntheses of a number of oligopeptides by classical means are reported. The peptides are designed as models for possible antigenic sequences among the Tyr/Glu sequences of a macromolecule widely used as antigen in immunogenetic studies, the multichain co-polymer, poly(Lys)  poly(Tyr, Glu)-poly(DL Ala). Preliminary studies on the biological activity of the oligopeptides synthesized show that only peptides with the N-terminal sequence H-Tyr-Glu- do compete with the co-polymer in a double antibody radioimmune assay, whereas other sequences do not.


Influenza: the agent.

Influenza: the agent. Less perplexing than its epidemic behavior is the nature of the influenza virion as such, which is now one of the best understood viruses in terms of morphology and replication. Among the aspects of the virus discussed in this article are the composition of its genome, the components of its envelope, and the probable molecular structure of its two surface proteins as well as their relative roles in antigenicity.


Cell culture factors influencing in vitro expression of mouse mammary tumor virus. Several cell culture factors were found to influence in vitro expression of mouse mammary tumor virus (MMTV) in the mouse adenocarcinoma cell line Mm5mt/c1. Cells were propagated in a variety of commercially available cell culture media to which dexamethasone (DXM) was added as a stimulator of MMTV production. Culture seeding density, culture medium type, and glucose concentration each influenced MMTV production when expressed on a per cell basis. Maximum cell growth occurred in cultures grown in RPMI-1640 medium containing insulin. Those media which provided good cell growth were not necessarily optimal for virus expression. Addition of insulin did not stimulate MMTV synthesis although dexamethasone alone was stimulatory in all media used; however, maximum MMTV expression was obtained when dexamethasone and insulin were used in concert. Equivalent levels of MMTV-specific cell membrane antigen, MMTV-specific protein, and virus particles were produced at incubation temperatures of 32 degrees, 34 degrees or 37 degrees C; however, higher levels of virus-related RNA-dependent DNA polymerase (RDDP) activity were recovered from cultures incubated at 32 degrees and 34 degrees C than at 37 degrees C. Decreased levels of RDDP were attributed to enzyme thermolability at 37 degrees C incubation.


[Diagnostic stain of helminth eggs (author's transl)]. A description is given of a diagnostic method for the staining of eggs and larvae of intestinal helminth in smears of both fresh and fixed stool samples. The contents of the eggs and larvae stain red, the background various shades of blue. The most contrasting staining was obtained with thin-walled eggs.


[Side effects of aminoglycosides: nephrotoxicity (author's transl)]. Animal experiments showed that all aminoglycosides cause similar toxic tubular and glomerular damage when investigated by qualitative morphology. Quantitative differences in the tubular nephrotoxicity of gentamicin, tobramycin, sisomicin, kanamycin, kanendomycin, amikacin, and butirosin can be demonstrated experimentally by evaluation of the excretion rates of tubular cells and urinary enzymes in rats. By this means dose-effect relationships were established resulting in varying reproduceable toxic threshold doses for each antibiotic, and thus in a scale of increasing nephrotoxicity. The aminoglycosides differed in their affinity to kidney tissue as measured by determination of the accumulating renal concentrations of the drugs at different times during multiple-dose administration. This had a modifying influence on excretion rates of cells and enzymes affecting the scale of toxicity in long-term studies. Comparative investigations on nephrotoxicity in rats and guinea pigs gave similar results. In addition, a study in man suggested that the test results of nephrotoxicity are not species-specific. For human therapy it is concluded that even more caution should be practised with the new aminoglycerides than with gentamicin in order to avoid renal damage.


Penicillin allergy studies by a modified red-cell-linked antigen-antiblobulin reaction. A red-cell-linked antigen-radioactive antiglobulin reaction (RCLA AR) is reported which may be combined with the conventional red-cell-linked antigen-antiglobulin reaction (RCLAAR) to measure antibodies of all five immunoglobulin calsses against the same determinant at the same time. The method has been applied to sera from penicillin-allergic patients and controls. Human erythrocytes coated with rabbit antibodies carrying penicilloyl groups were incubated with the serum to be tested, then washed. 125I-labelled anti-human IgE was added and, after further incubation and washing, the radioactivity retained by the cells was counted. This gives a measure of the specific IgE in the test sample. Other aliquots of the coated cells were treated with specific antiglobulin reagents in the usual manner of the RCLAAR in order to determine antibodies of other classes. The sensitivity for IgE was similar to that of the radioallergosorbent test. Information was also obtained about specific IgG subclasses.


Fetuin. In vitro effect on sheep erythrocyte rosette formation with human T lymphocytes. Peripheral blood lymphocytes from six healthy control subjects were incubated in vitro with fetuin and enumerated for sheep erythrocyte rosette forming T cells. Significant enhancement of rosette-forming T lymphocytes was observed in a dose-related manner. The physical presence of fetuin is not required for this effect. Incubation of lymphocytes with human AB serum has no significant effect on the rosette formation. This enhancing effect of fetuin appears to be secondary to alteration in T lymphocyte surface receptors for sheep erythrocytes.


Antibodies against hydroxyethylstarch produced in rabbits by immunization with a protein-hydroxyetylstarch conjugate. Antibodies (ABS) against hydroxyethylstarch (HES) were raised in rabbits by immunization with HES-bovine serum albumin conjugate. ABS against HES were demonstrable by gel diffusion, passive hemagglutination (PH) and passive cutaneous anaphylaxis (PCA); specificity was confirmed by inhibition of PH and neutralization of PCA. No ABS against starch could be induced in comparative experiments. Non-immunogenicity of starch was attributed to its structural similarity with glycogen, widely distributed in animal species. Absence of cross-reactivity of anti-HES ABS with starch, amylopectin and glycogen and strong reactivity with 2-hydroxypropylstarch (DS = 0.65) and HES (DS = 0.7-1.2) indicate that hydroxyethyl substitution created antigenic determinants and conferred new immunochemical identity on the modified starch molecule. Anti-HES ABS represent specific analytical tools for the identification and quantitation of HES in biological material.


Immunological activities of rat lymphocytes. I. Mitogenic responses and surface markers of lymphocytes from normal, thymectomized and B rats. Lymphocytes from the peripheral blood, thoracic duct, spleen and lymph nodes of normal, thymectomized (TX) and thymectomized lethally irradiated marrow reconstituted (TXBM or B rats) Lewis rats were studied for their ability to proliferate in vitro in the presence of Con A or PHA. At the same time the lymphoid tissues of these animals were examined for the presence of B cells or T cells by immunofluorescence staining with antiserums to rat immunoglobulins or rat brain antigens (ARBS), respectively. The specificity of ARBS fort T cells was first established in both cytotoxicity and immunofluoresecnce studies with thymocytes as well as pruified T and B cells. In various lymphoid tissues from TX and B rats, cells carrying brain antigens were found which were unable to respond to Con A and PHA. Thymectomy has a lesser effect on the response to Con A that to PHA; this was particularly true in the thoracic duct where, following thymectomy, lymphocytes continued to respond to Con A but lost their ability to respond to PHA. This suggests that the response to PHA and Con A may belong to different cells' subsets but that the ability to respond to either one of these mitogens may be impaired in cells carrying known T cell markers.


Determination of alpha-fetoprotein by a new paper disc radioimmunoassay of the sandwich type. Determination of alpha-fetoprotein (AFP) in the 1- to 500-mug/litre range has been shown to be of diagnostic value in a variety of clinical settings, in particular, in prenatal diagnosis and the monitoring of pregnancies. A modified paper disc sandwich radioimmunoassay (sandwich RIA) is described which meets the essential criteria for large-scale assaying for AFP. The sandwich RIA is based upon anti-AFP antiserum, which is available commercially, and thus purified AFP is not essential. The sandwich RIA (1) is simple to run and gives results within 24 h; (2) has a comparatively broad range of sensitivity; (3) requires no apparatus other than a gamma-scintillation counter, and (4) has an acceptable level of accuracy. We have used the described sandwich RIA for AFP in routine obstetrical practice for over 2 years. Our experience, encompassing more than 15,000 samples, has been most satisfactory.


Flour allergy in bakers. I. Identification of allergenic fractions in flour and comparison of diagnostic methods. Extract of wheat flour obtained by extraction, centrifugation and dialysis was immunochemically characterized by quantitative immunoelectrophoresis using rabbit antibodies. The analysis revealed wheat flour to be composed of 40 antigens, some of which were immunologically partially identical with antigens of rye flour and of common grass pollen. Furthermore, antigens of the gliadin fraction of wheat flour were identified. 25 bakers with allergic complaints working in and around Copenhagen were clinically tested with wheat flour and related extracts. Among 13 bakers with respiratory complaints (asthma and/or rhinitis), 11 showed positive reactions to wheat flour extract when tested in histamine release from basophil leukocytes radioallergosorbent test and skin test, whereas further 2 were positive in the basophil test only. The ability of the IgE of individual sera to adsorb to the individual antigens of wheat flour was examined by means of crossed radioimmunoelectrophoresis. On the basis of these results, individual allergenic components of wheat flour were identified, three of these with comparatively high affinity and frequency.


Sequential release of histamine and 5-hydroxytryptamine during pinnal anaphylaxis in the mouse. Histamine and 5-HT have been demonstrated to be involved in pinnal anaphylaxis in mice. Each predominates at different stages during the 30-min development of the reaction. The main mediator of the early stage of the reaction (0-10 min) is histamine, that of the middle stage (10-20 min) is 5-HT, and that of the final 20- to 30-min stage is, again, histamine. The source and mechanism of release of the 5-HT and the late histamine are, as yet, unknown but under investigation.


The allergenic structure of allergen M from cod. I. Tryptic peptides of fragment TM 1. In attempts to locate the allergenic active sites of fragment TM 1 of allergen M from cod, the allergenicity of the tryptic hydrolysis peptides was examined both in vivo and in vitro. A duodecapeptide (residues No. 33-44) was tentatively suggested to incorporate at least one allergenic determinant. All the tryptic peptides were less active than allergen M as shown by the in vivo titration experiments. The activity of the peptides was not due to contamination of intact allergens.


Inhibition of reagin-mediated PCA reactions in monkeys and histamine release from human leukocytes by human IgG4 subclass. Human myeloma proteins of IgG4 subclass in contrast to myeloma proteins IgG1, IgG2 and IgG3, were capable of blocking PCA reactions in monkeys mediated by human reaginic antibodies of IgE class. In addition to IgE, IgG4 myeloma protein was also capable of sensitizing leukocytes from normal individuals and gave histamine release (HR) upon challenge with anti-human IgG4. Leukocytes from 11 allergic individuals and from 9 normal subjects sensitized with the serum of allergic patients, were capable of releasing histamine with anti-human IgG4, anti-human IgE, and the specific allergen. No response was obtained with anti-human IgG1 and IgG3 sera. Leukocytes from the normal individuals released histamine from 3 to 20% with anti-human IgG4 and from 6 to 30% with anti-human IgE. Moreover, normal leukocytes sensitized with IgG4 myeloma protein or a serum of an allergic patient heated at 56 degrees C for 2 h, released a significant amount of histamine on challenge with anti-human IgG4 whereas no response was obtained with anti-human IgE. The biological role of human IgG4 in immediate hypersensitivity reactions is discussed in relation to human IgE.


Development of radioimmunoassay for pregnancy-specific beta1-globulin and its measurement in serum of patients with trophoblastic and non-trophoblastic tumours. Levels of human pregnancy-specific or trophoblast-specific beta1-globulin (TBG) were measured by double-antibody radioimmunoassay (RIA) in 103 patients with trophoblastic tumours and in 114 patients with a variety of non-trophoblastic tumours. The sensitivity of RIA for TBG was about 1 ng per ml. Twenty healthy adult females and males had levels below 1 ng per ml. Using the RIA for measurement of TBG in sera which were selected from immunodiffusion-TBG-negative samples, we detected elevated TBG levels in 76.7% of cases with trophoblastic tumours and in 15% of different cases with non-trophoblastic malignancies. The diagnostic and prognostic significance of the measurement of TBG levels by RIA in patients with trophoblastic tumours is discussed.


Mouse mammary tumor virus production stimulated by hormones and polyamines in cells grown in semi-synthetic in vitro conditions. Hormonal regulation of mouse mammary tumor virus (MMTV) synthesis was studied in the CCL-51-SF cell subline derived from the Sykes' mammary tumor cell line CCL-51 and adapted to grow in semi-synthetic in vitro conditions. The virus was quantitated by measuring the supernatant reverse transcriptase activity in exogenous reaction using poly (rA)-oligo (dT) and poly (rC)-oligo (dG) as template/primers. The cells produced a low but significant amount of virus in the absence of any hormones and serum proteins. The synthetic glucocorticoid dexamethasone increased production considerably, up to 100-fold. Pretreatment of CCL-51-SF cells with serum or 5-bromodeoxyuridine, BrdUrd, partly reduced the stimulation by dexamethasone of MMTV production. Insulin and prolactin alone or in combination had no stimulating effect on spontaneous MMTV synthesis and cell growth. Prolactin, and more efficiently the prolactin-insulin combination, enhanced the MMTV production stumulated with dexamethasone. Insulin alone remained without effect. The polyamine spermidine, but not spermine, increased the MMTV production over the control by a factor of 2. Polyamines did not influence cell replication at the concentrations used.


Improvement of intractable idiopathic epilepsy with chlormadinone acetate. A case report. A 10-year-old boy with idiopathic epilepsy refractory to treatment from age 4 and with clinical signs of hyperandrogenism was treated with chlormadinone acetate (24 mg/day) during two alternate periods of 3 months. The administration of chlormadinone was associated with a marked clinical and electroencephalographic improvement as well as a concomitant decrease in the levels of plasma testosterone and delta4 androstenedione. No changes in plasma gonadotropins were observed. Placebo had no effect neither in seizures nor in the concentration of hormones in the plasma. Since chlormadinone acetate is not a very potent gonadotropin inhibitor in males, it is suggested that the clinical and electroencephalographic improvement associated with the use of this compound, might be the consequence of a decrease in the plasma levels of androgens, mainly testosterone.


Histochemical studies of mucosubstances and lipids in normal human oesophageal epithelium. The histochemistry of lipids and mucosubstances in normal human oesophageal epithelium were studied in biopsies obtained from 24 patients undergoing investigations of the upper gastro-intestinal tract. Neutral fat droplets, 1-2 mum in diameter, were present in all layers, the greatest number being in the functional layer cells. No unsaturated lipids or fatty acids were demonstrable. Much glycogen was present in the cytoplasm of the prickle and functional cell layers as demonstrated by PAS (and diastase digestion) techniques. The intercellular space of the most superficial of the functional cell layer contained neutral and sialic acid-rich acid mucopolysaccharides. These may be important in protecting the epithelium against physical and chemical trauma.


The Alcian blue and combined Alcian blue--Safranin O staining of glycosaminoglycans studied in a model system and in mast cells. Polyacrylamide films containing different glycosaminoglycans have been applied to the study of the Alcian Blue and combined Alcian Blue--Safranin O staining procedures. It was found that the polyacrylamide matrix can be interpreted as some kind of 'barrier' around the substrate molecules, a situation which can be compared to a certain extent with what occurs in situ, where complex protein molecules can likewise form a 'barrier'. The Alcian Blue staining of the model films was found to follow the Lambert-Beer law. The time to reach optimal dye binding depended on the concentration of the glycosaminoglycan enclosed in the model films and on the concentration of Alcian Blue in the dye solution. Lowering the pH of the dye solution appeared to increase the rate of staining. Optimal staining of model films in the presence of salt or urea was not possible, because under these conditions the pores of the polyacrylamide matrix became blocked. Alcian Blue was found to bind irreversibly to the glycosaminoglycan molecules enclosed in the polyacrylamide films. The results of the combined Alcian Blue-Safranin O staining applied to model films appeared to be highly dependent on the amount of Alcian Blue bound to the glycosaminoglycan in the first step of the double staining procedure. No specific differences were noticed between the behaviour of the different glycosaminoglycan-Alcian Blue complexes towards the Safranin O binding in the next step. As the theoretical basis for the application of the combined Alcian Blue-Safranin O staining was also found not to be completely valid, the conclusion was reached that this double staining cannot be used for the histochemical identification of glycosaminoglycans. The colour retained by a certain glycosaminoglycan-containing part of the specimen only delivers information about the accesibility of that part for Alcian Blue.


An investigation into the hypothesis of shifting neuronal relationships during development. The projection of ganglion cells from the centre of the eye to the tectum has been followed during development in Xenopus by an autoradiographic technique. By comparing the caudal limit of this projection with a fixed line of cells labelled with tritiated thymidine the projection has been shown to shift caudally during development. This indicates a less rigid form of neuronal specificity than has previously been accepted.


Argyrophilic cells in the larynx of the guinea-pig demonstrated by the method of Grimelius. Argyrophilic cells with branching processes are shown in both the surface epithelium and in the glands in the larynx of adult guinea-pigs using the Grimelius silver technique after GPA fixation. When Bouin's fixative or neutral formalin were used as fixatives argrophilic cells could not be identified.


Stability and subunit structure of human alpha2-macroglobulin. The molecular weights of alpha2-macroglobulin and its non-covalent subunits have been determined by equilibrium centrifugation. The secondary structure of the native and the thermally denatured molecules has been analyzed by circular dichroic measurements. In contrast to most proteins the thermally denatured form contains a slightly more highly organized polypeptide chain than the native form. The relaxation time of the native protein, as determined by fluorescence polarization measurements, indicates that alpha2-macroglobulin is composed of domains smaller than that of the two subunits. The transitions in acid, alkali, and at high temperatures have been explored in order to establish the pH and thermal range of stability of alpha-macroglobin.


Treatment of an encephalitogenic peptide from guinea pig myelin basic protein with alpha-protease and thermolysin. Isolation of fragments and determination of cleavage sites. Two peptic fragments (residues 37-88 and 43-88) of guinea pig myelin basic protein which are capable of inducing experimental allergic encephalomyelitis in Lewis rats were cleaved to shorter fragments with alpha-protease (Crotalus atrox proteinase, EC 3.4.24.1) and thermolysin (EC 3.4.24.4). The fragments were isolated, purified, and identified by amino acid composition and NH2- and COOH-terminal residues. The time courses of the reactions, monitored by thin layer electrophoresis of the digests, showed that alpha-protease cleaves peptide (43-88) initially at the Pro(71)-Gln(72) bond, and that the product peptides are subsequently attacked at the Arg(63) -Thr(64), Ser(74)-Gln(75), Arg(78)-Ser(79), and Ser(76)-Gln(80) bonds. No significant cleavages occurred at the -Leu, -Val, and -Ala bonds. These results are in striking contrast to those obtained previously by others workers with other peptide substrates, where selective cleavage at hydrophobic residues occurred. Thermolysin was found to attack peptide (37-88) at the Phe(42)-Phe(43) bond very rapidly; the product peptides were subsequently attacked at the His(60)-Ala(61), Ser(38)-Ile(39)-Tyr(67)-Gly(68), and Pro(84)-Val(85) bonds. These cleavages are compatible with the known specificity of this enzyme. Several of the fragments prepared with these two enzymes, peptides (43-71), (61-88), (75-88), and (72-84) have been used in other studies to locate the encephalitogenic site in the parent peptic peptide.


Cancer-associated serum galactosyltransferase activity. Demonstration in an animal model system. Two different lines of solid tumors were produced in outbred hamsters by subcutaneous injection of polyoma transformed BHK cells. Growth of the tumors correlated with the appearance in serum of an electrophoretically distinct peak of galactosyltransferase: NeuAc-, Gal-free fetuin acceptor activity on polyacrylamide gels. This slow moving peak of enzyme activity (GT-HH) was detected before solid tumors could be grossly observed and the amount of activity in this peak was also found to be linearly related with growth of the tumor. GT-IIH was not detectable in control animals and separated from a faster migrating major area of serum galactosyltransferase activity (GT-IH) found in sera of both control and tumor-bearing hamsters. These two activities were shown to maintain their respective mobilities on re-electrophoresis. Solubilized enzyme derived from excised tumors demonstrated an electrophoretic mobility on polyacrylamide gels identical to that for GT-IIH present in serum from tumor-bearing animals. In contrast, enzyme activity solubilized from livers of both control or tumor-bearing hamsters showed a mobility similar to that of the faster moving serum galactosyltransferase enzyme activity, i.e. GT-IH. In addition, medium derived from nonconfluent BHKpy cells in tissue culture contained galactosyltransferase activity which co-electrophoresed with the slower migrating characteristics of galactosyltransferase activities derived from serum (control and tumor-bearing), solid tumors, liver and BHKpy cells in tissue culture were compared. All kinetic properties were similar with the exception that the Km UDP-galactose of GT-IIH (1.0 X 10(-5) M) was half that of GT-IH (2.0 X 10(-5) M).


Arrangement of high molecular weight associated proteins on purified mammalian brain microtubules. The arrangement of the high molecular weight proteins associated with the walls of reconstituted mammalian brain microtubules has been investigated by electron microscopy of negatively stained preparations. The images are found to be consistent with an arrangement whereby the high molecular weight molecules are spaced 12 tubulin dimers apart, i.e., 960 A, along each protofilament of the microtubule, in agreement with the relative stoichiometry of tubulin and high molecular weight protein. Molecules on neighbouring protofilaments seem to be staggered so that they give rise to a helical superlattice, which can be superimposed on the underlying tubulin lattice. In micrographs of disintegrating tubules there is some indication of lateral interactions between neighbouring high molecular weight molecules. When the microtubules are depolymerized into a mixture of short spirals and rings, the high molecular weight proteins appear to remain attached to their respective protofilaments.


A serum factor requirement for the passage of cultured Vero cells through G2. When Vero cells, a line derived from and African Green Monkey kidney, are grown under conditions where the saturation density is limited by serum, they deplete the growth medium of a factor necessary for cell division. The factor is a component of serum. When Vero cells are plated at low density (2 X 10(4)/cm2) in this depleted growth medium (after dialysis against serum-free Dulbecco's Modified Eagle's Medium) they initiate an unbalanced program of growth. Protein synthesis proceeds at the same rate as parallel cells in fresh serum, and and the cells accumulate protein as a function of time. DNA synthesis is also initiated in these cells, and the amount of DNA per cell increases for the next four days plating. However the cells quickly stop dividing. Measurements of DNA per cell using microspectrofluorometry show that the cells are accumulating in the late S and G2 period during this time. Thus we conclude that these cells cannot pass through a transition point in G2. When fresh serum is added to cells after three days in depleted growth medium, they divide before they begin to synthesize DNA. This further confirms that they are in late S and G2. Cell division is promoted in Vero cells in depleted growth medium by bovine fetuin, and to a lesser extent by bovine albumin. Cell division is not promoted by insulin, hydrocortisone, dexamethasone, linolenic acid, calcium, and typsin inhibitor form ovomucoid. From these data we conclude that transit through G2 requires the prescence of an extracellular factor.


Alterations in differentiation and pyrimidine pathway enzymes in 5-azacytidine resistant variants of a myoblast line. 5-azacytidine at concentrations higher than 5 muM inhibited the differentiation of a rat myoblast line in vitro. It was also somewhat cytotoxic at this level. Variants resistant to the cytotoxic effect of 5-azacytidine were obtained which were simultaneously unable to differentiate into myotubes and exhibited altered morphology. These characteristics were retained by the variants when subcultured in the absence of the drug for over 700 generations. Several of the azacytidine resistant cells were more susceptible than the parental line to the lethal action of 5-bromodeoxyuridine and adenosine, but not that of cytosine arabinoside, ouabain or 8-azaguanine. The variants were capable of transporting uridine, thymidine and 5-azacytidine. The uridine kinase activity was one-half to one-third of than in the parental cells but it was not missing completely in any of the variants. Two independently isolated variants selected for detailed study showed a 2- to 3-fold increase in the activity of orotidylic acid decarboxylase. This enzyme in the variants in contrast to that of the parental cells was completely insensitive to the inhibitory effect of a nucleotide generated from ATP and 5-azacytidine in cell extracts (probably 5-azacytidine monophosphate). These observations point to the possibility the 5-azacytidine resistance arises in myoblasts due to an alteration of the components of two target pathways of this drug, viz., the de novo  pyrimidine pathway and an undefined sequence leading to the synthesis of membrane components.


Study of the pituitary-thyroid functions at high altitude in man. The alterations in serum levels of T3, T4, TSH and TBG, TSH response to 100 mug iv TRH, and urinary excretion of T3 and T4 were studied in 8 healthy men at sea level (SL), on days 1, 2, 4, 8 and 16 after arrival by air at high altitude (3,700 m, HA), and during days 5 to 7 after their return to SL. No significant alterations in serum levels of TSH and TBG or TSH response to TRH were observed during exposure to HA or on return to SL. There was, however, an acute elevation in both serum total T3 and T4. Serum total T3 from a mean basal+/-SE value of 128+/-13 ng/dl increased to 320+/-18 on day 1 and remained significantly elevated at 225+/-48 up to day 8 after arrival at high altitude. Similarly serum total T4 increased from basal level of 9+/-0.92 mug/dl to 15.2+/-1.2 and remained elevated till day 16 and it was 11+/-1.19 mug/dl during days 5 to 7 after return to SL. The urinary excretion of both T3 and T4 was decreased. These changes perhaps were the result of complex physiologic adjustments on acute exposure to high altitude, like shrinkage of the T3 and T4 distribution pools, altered binding capacities of thyroid hormones binding proteins, and a reduction in clearance of thyroid hormones from the plasma compartment; and were probably not suggestive of an enhanced thyroid activity. Their actual significance in high altitude adaptation in man is not clearly understood.


Hypothyroidism in an area of endemic goiter and cretinism in Central Java, Indonesia. In an area of severe endemic goiter in Central Java, Indonesia, clinical overt or mild hypothyroidism appeared to be present in 7 out of 20 cretins and also in 12 out of 94 non-cretinous subjects, all 5-20 years of age, living in the village of Sengi. Hypothyroidism was not found in a control group of 70 subjects of the same age living in Londjong just outside the edemia. In hypothyroid subjects the plasma PBI-concentration was 0.98+/-0.32 mug/100 ml (mean+/-SD) vs. 2.72+/-1.24 mug/100 ml in euthyroid subjects from Sengi and 4.86+/-0.80 mug/100 ml in controls from Londjong. Values for T3 were 56.3+/-3.17 ng/100 ml in hypothyroids, 140.5+/-38.5 ng/100 ml in euthyroids from Sengi and 121.6+/-27.4 ng/100 ml in controls. The TSH levels (geometric mean and range) in these 3 groups were, respectively, 210.1 (108.0-342), 15.6 (3.0-372) and 4.1 (0.8-7.0) muU/ml. The differences between the mean concentration of PBI, T3 and TSH in the hypothyroid and euthyroid groups were highly significant (P less than 0.001). These data strengthen the clinical diagnosis of hypothyroidism in cretins as well as in non-cretinous subjects. All hypothyroid subjects had a PBI less than 1.8 mug/100 ml and T3 less than 120 ng/100 ml and TSH greater than 100 muU/ml. In 8 hypothyroid subjects, restudied 18 months after iodized oil injection, hypothyroidism was either corrected or markedly improved. It therefore appears that iodine deficiency per se in postnatal life may lead to (juvenile) hypothyroidism, which can be corrected by iodine therapy. Our findings have implications for the definition and diagnosis of endemic cretinism. Not all hypothyroid subjects in an area of endemic iodine deficiency should be classified as cretins.


Efferent projections of the main and the accessory olfactory bulb in the tree shrew (Tupaia glis). The projections of the main and the accessory olfactory bulb in the tree shrew (Tupaia glis) have been analyzed with anterograde degeneration and autoradiographic methods for identifying axonal projections, and with the horseradish peroxidase method for identifying the distribution of neurons from which these projections originate. The cytoarchitectonic features of the paleocortical areas which receive projections from the main and the accessory olfactory bulb have also been described. The efferent projections of the accessory olfactory bulb are distributed to the bed nucleus of the accessory olfactory tract, the medial amygdaloid area, the posteromedial cortical amygdaloid area, and to the caudal portion of the bed nucleus of the stria terminalis. In contrast, the efferent projections of the main olfactory bulb are distributed to the anterior olfactory nucleus, the tenia tecta, the olfactory tubercle, the pyriform cortex, the anterior cortical amygdaloid area, the posterolateral cortical amygdaloid area, and to the lateral entorhinal cortex. These observations are consistent with the notion that the olfactory system can be divided into at least two major subsystems: one related to the vomeronasal organ and accessory olfactory bulb, and another related to the main olfactory organ and main olfactory bulb. The paleocortical areas receiving olfactory projections have three basic layers: a superficially positioned plexiform layer (layer I), a pyramidal cell layer (layer II), and a polymorphic cell layer (layer III). The projections of both the main and the accessory olfactory bulb terminate in the outer portion of the plexiform layer (sublamina Ia). Sublamina Ia contains the distal segments of dendrites which originate from a heterogeneous population of neurons located in layer II and, to a lesser extent, layer III. Although the efferent projections of the main and the accessory olfactory bulb are segregated, evidence for a more refined topographical organization within these projections was not obtained. However, the distribution of retrogradely labeled neurons in the main olfactory bulb, following injections of horseradish peroxidase into its various paleocortical targets, indicates that the olfactory projections to these areas may not all originate from the same population of cells.


Bilateral ganglion cell branches in the normal rat: a demonstration with electrophysiological collision and cobalt tracing methods. The application of microelectrode recording and anatomical tracing methods to the subcortical optic projections of the normal rat shows that the uncrossed pathway from the retina of this mammal is substantially composed of branches of crossed axons. In the recording experiments, paired stimuli to the optic tracts produces an attenuated antidromic response in the optic nerve which is best explained by the collision of impulses which travel in branches of the same parent axon. Cobalt injection of one optic tract results in retrograde filling of axons in the entire contralateral optic nerve and filling of axons in restricted regions of the ipsilateral optic nerve. This procedure also results in the filling of axons in the opposite optic tract. The locations of the filled axons in the opposite tract correspond to the positions of crossed and uncrossed projections from the temporal retinae: ventrolateral and dorsomedial, respectively. The position of the temporal retinal projections in both optic tracts was determined by applying silver degeneration methods after small lesions of the retina.


The organization of the facial nucleus of the brush-tailed possum (Trichosurus vulpecula). The facial nucleus of the brush-tailed possum has been studied using Nissl staining and the horseradish peroxidase (HRP) retrograde tracing technique. In Nissl stained sections the nucleus is seen to comprise five distinct subnuclei. Injections of HRP into individual facial muscle groups have shown that these subnuclei reflect the peripheral innervation pattern of efferents from this nucleus. Although in most cases, injection of HRP into a single facial muscle group resulted in the labelling of neurons in more than one facial subnucleus, the following subnuclei were most completely labelled subsequent to intramuscular injection of HRP: the dorsal intermediate subnucleus was labelled with HRP reaction product following injection of m. auricularis anterior; the middle intermediate subnucleus  was labelled following injection of the muscle underlying the malar vibrissae; the ventral intermediate subnucleus was labelled following injection of the m. mentalis; the medial subnucleus was labelled following injection of the m. auricularis posterior; the lateral subnucleus was labelled following injection of the m. nasolabialis with HRP. In general there is a mediolateral representation in the facial nucleus of neurons innervating facial muscle groups which are found in anteroposterior succession along the head of the animal. Muscle groups found in dorsoventral succession on the animal are represented dorsoventrally in the facial nucleus.


An autoradiographic study of the organization of the efferent connections of the hippocampal formation in the rat. The efferent connections of the hippocampal formation of the rat have been re-examined autoradiographically following the injection of small quantities of 3H-amino acids (usually 3H-proline) into different parts of Ammon's horn and the adjoining structures. The findings indicate quite clearly that each component of the hippocampal formation has a distinctive pattern of efferent connections and that each component of the fornix system arises from a specific subdivision of the hippocampus or the adjoining cortical fields. Thus, the precommissural fornix has been found to originate solely in fields CA1-3 of the hippocampus proper and from the subiculum; the projection to the anterior nuclear complex of the thalamus arises more posteriorly in the pre- and/or parasubiculum and the postsubicular area; the projection to the mammillary complex which comprises a major part of the descending columns of the fornix has its origin in the dorsal subiculum and the pre- and/or parasubiculum; and finally, the medial cortico-hypothalamic tract arises from the ventral subiculum. The lateral septal nuclei (and the adjoining parts of the posterior septal complex) constitute the only subcortical projection field of the pyramidal cells in fields CA1-3 of Ammon's horn. There is a rostral extension of the pre-commissural fornix to the bed nucleus of the stria terminalis, the nucleus accumbens, the medial and posterior parts of the anterior olfactory nucleus, the taenia tecta, and the infralimbic area, which appears to arise from the temporal part of field CA1 or the adjacent part of the ventral subiculum. The projection of Ammon's horn upon the lateral septal complex shows a high degree of topographic organization (such that different parts of fields CA1 and CA3 project in an ordered manner to different zones within the lateral septal nucleus). The septal projection of "CA2" and field CA3 is bilateral, while that of field CA1 is strictly unilateral. In addition to its subcortical projections, the hippocampus has been found to give rise to a surprisingly extensive series of intracortical association connections. For example, all parts of fields CA1, CA2 and CA3 project to the subiculum, and at least some parts of these fields send fibers to the pre- and parasubiculum, and to the entorhinal perirhinal, retrosplenial and cingulate areas. From the region of the pre- and parasubiculum there is a projection to the entorhinal cortex and the parasubiculum of both sides. That part of the postsubiculum (= dorsal part of the presubiculum) which we have examined has been found to project to the cingulate and retrosplenial areas ipsilaterally, and to the entorhinal cortex and parasubiculum bilaterally.


The olivocerebellar projection in the cat studied with the method of retrograde axonal transport of horseradish peroxidase. IV. The projection to the anterior lobe. Following injections of horseradish peroxidase (HRP) in the cerebellar cortex of the anterior lobe of the cat, the distribution of labeled cells in the inferior olive was mapped. The findings largely confirm those made previously in studies of olivary retrograde cell loss following cerebellar ablations (Brodal, '40b). In addition, they reveal further olivary areas projecting onto the anterior lobe, and permit a more detailed analysis of the pattern in this projection. Concerning major points the results are in agreement with physiological studies by Armstrong et al. ('74). They bring supporting evidence for a longitudinal zonal pattern in the anterior lobe (fig. 6C). The middle zone of the vermis receives its fibers from a large central area in the caudal half of the medial accessory olive, a lateral zone of the vermis from the lateral half of the dorsal accessory olive. Both olivary areas project to the corresponding cerebellar zone throughout lobules V-I. The lateralmost part of the anterior lobe (lobules IV-V) receives afferents from an area in the dorsal lamella of the principal olive. The intermediate part of lobules IV-V receives afferents from the medial half of the dorsal accessory olive and from an area in the rostral half of the medial accessory olive. There is suggestive evidence that the latter projects to a middle zone, the former to a medial and a lateral zone within the intermediate part as found physiologically. Conclusions concerning projections to the intermediate part of lobules III-II could not be made. The findings in this and preceding studies with the HRP-method show that the concept of a longitudinal pattern in the cerebellum is scarcely generally valid of the entire olivocerebellar projection. Within the projections of the lateral half of the dorsal accessory olive and the area in the rostral part of the medial accessory olive there appears to be a topical relation with the folial pattern in the anterior lobe. An analysis of the findings with reference to the afferents traced anatomically to the various olivary areas permits some conclusions as to the functional role of the olivary areas. Comparison with Oscarsson's ('73) diagram of the sites of termination of two of the spinal-olivary pathways (his DF-SOCP) and VF-SOCP) permits an anatomical explanation as concerns the projections to the vermis, while correlations as concerns the intermediate part are less satisfactory.


Brain stem afferents to the fastigial nucleus in the cat demonstrated by transport of horseradish peroxidase. Although retrograde and anterograde degeneration studies have provided important information concerning brain stem afferents to the fastigal nucleus (FN), these data may be incomplete and should be confirmed by axonal transport methods. Attempts were made to inject horseradish peroxidase (HRP) unilaterally into the FN in a series of adult cats. Animals were perfused with dextran and a fixative solution of paraformaldehyde and glutaraldehyde in 0.1 M phospate buffer. Representative sections were treated by the Graham and Karnovsky ('66) method. Selective HRP injections in one FN resulted in retrograde transport of the marker to Purkinje cells of the ipsilateral vermis and distinctive appendages of the contralateral medial accessory olivary (MAO) nucleus (nucleus beta and the dorso-medial cell column). Retrograde transport of the label was found bilaterally in cells of the medial (MVN) and inferior (IVN) vestibular nuclei, in cell group x and in the nucleus prepositus (PP). Labeled vestibular neurons, most numerous in MVN, were identified in dorsal, caudal and lateral regions, with a slight ipsilateral preponderance. Only a few neurons in caudal, dorsal and lateral regions of the IVN were labeled and none of these included cells of group f. Labeled cells in the caudal third of PP were greatest ipsilaterally. Rostral and caudal injections of FN labeled smaller numbers of cells in MVN, IVN, cell group x and PP. HRP injections of FN and portions of lobules VIII and IX resulted in bilateral retrograde labeling of larger numbers of cells in MVN, IVN and cell group x, and ipsilateral labeling of cells in group y and the interstitial nucleus of the vestibular nerve. Injections of HRP into basal folia of lobules V and VI resulted in retrograde transport of the marker to cells of the medial and dorsal accessory olivary nuclei contralaterally, and to cells of the ipsilateral accessory cuneate nucleus. Transport of label injected into portions of the pyramis was detected in parts of the contralateral MAO and bilaterally in parts of the pontine and reticulotegmental nuclei. This study suggests that the principal afferents of the fastigial nucleus arise from: (1) Purkinje cells of the ipsilateral vermis, (2) restricted portions of the contralateral MAO (nucleus beta and dorsomedial cell column), (3) portions of the MVN and IVN (bilaterally) and (4) caudal parts of the PP. Secondary vestibular inputs to the fastigial nucleus probably are relayed mainly by Purkinje cells in the cerebellar cortex.


The vestibular complex of the American opossum didelphis virginiana. II. Afferent and efferent connections. We have demonstrated the connectivity of the opossum's vestibular nuclei using degeneration, autoradiographic and horseradish peroxidase techniques and have found it to be generally comparable to that reported for the cat. Apart from the primary input described in Part I of our study, the cerebellum provides the major source of afferent connections to the vestibular complex. Axons from the cerebellar cortex distribute mainly to vestibular areas which receive no primary afferent projections, e.g., the dorsal part of the lateral vestibular nucleus, the dorsolateral margin of the inferior vestibular nucleus as well as cell groups comparable to "f" and "x." In contrast, fastigial fibers show considerable overlap with primary vestibular input, particularly in the ventral part of the lateral nucleus, the central part of the inferior nucleus and the medial nucleus. Axons of fastigial origin also distribute to the superior vestibular nucleus, to subnuclei "f" and "x" and to the parasolitary region. Although spinal fibers are diffuse within the main vestibular nuclei, they ramify quite densely within subnucleus "x." Most of the spinovestibular projection appears to arise in the cervical spinal cord of the opossum. Ipsilateral connections from the interstitial nucleus of Cajal and surrounding areas end predominantly, but not exclusively, in the medial vestibular nucleus. A crossed midbrain projection has been verified from the red nucleus to cell group "x" and the lateral part of the inferior nucleus, as well as to an area possibly comparable to cell group "z," as described for the cat. In Part I of our study we have shown that the major targets of primary vestibular fibers are the central part of the superior nucleus, a portion of the parabrachial complex possibly comparable to subnucleus "y"," the ventral part of the lateral nucleus and the medial nucleus. All of these primary zones give rise to fibers supplying extraocular nuclei and surrounding areas (present study). The ascending midbrain fibers from the superior nucleus end mainly ipsilaterally, whereas those from the putative subnucleus "y" and the medial vestibular nucleus distribute contralaterally for the most part. Although the dorsal part of the lateral vestibular nucleus has no primary vestibular input, it does receive a major projection from the cerebellar cortex. This same region of the lateral nucleus projects to the spinal cord, but not to extraocular nuclei. The ventral part of the lateral nucleus, and perhaps the medial nucleus, also relay to the spinal cord. Additional projections from all vestibular nuclei to the reticular formation provide indirect routes through which the vestibular nuclei can potentially effect multiple systems, including those innervating the spinal cord. Finally, commissural vestibular connections of the opossum are shown to arise within all four major nuclei.


An autoradiographic investigation of the subcortical visual system in chimpanzee. Based on one adult chimpanzee monocularly injected with radioactive proline, retinofugal fibers were found to terminate bilaterally in the suprachiasmatic, pregeniculate, lateral geniculate, olivary, pretectal and lateral terminal nuclei, and the superior colliculi; the existence of a dorsal terminal nucleus of the accessory optic system is in doubt. In the ipsilateral geniculate nucleus, the fibers terminate in layers 2, 3 and 5; in the contralateral nucleus, they end in layers 1, 4 and 6. Midway through the geniculate nucleus, layers 3 and 4 split medially into two daughter layers each. In the superior colliculi, most of the retinal terminals are aggregated superficially in a band located in the stratum griseum superficiale. The contralateral band is interrupted by gaps; the ipsilateral band has fewer gaps, is slightly thicker and located more deeply. There is a limited second tier of terminals in the contralateral superficial gray.


The substantia nigra of the rat: a Golgi study. Three variants of the Golgi method were employed to examine the cell types, their dendritic fields and organization and axonal trajectories within the substantia nigra of albino and hooded rats. In both sagittal and coronal sections, large, medium and small neurons were classified on the basis of soma size, extent of dendritic fileds and dendritic caliber. In general nigral cells have three to five primary dendrites that branch relatively infrequently. Some dendrites of all cell types have thinly scattered spines or varicosities. Small cells, found in all areas of the nucleus, have thin dendrites and small, nondirectional dendritic fields. These are considered to be interneurons. The medium cells found in pars compacta, presumed to be the dopaminergic cells of the nigroneostriatal pathway, send long dendrites into pars reticulata perpendicular to the course of pars compacta. In addition, these cells have a number of dendrites which remain in pars compacta. These cells have axons that run medio-dorsally. No axon collaterals were detected. Both large and medium cells are found in pars reticulata. Cells in the dorso-medial aspect of pars reticulata orient rostro-caudally and roughly perpendicular to the course of pars compacta, while cells in the peripeduncular area show a strict orientation which is parallel to the crus cerebri. Some pars reticulata cells emit axon collaterals while others remain unbranced for their observable lenght. Both large and medium cells are also seen in pars lateralis. These cells send long dendrites ventrally into pars reticulata where they run parallel to the crus cerebri, while some shorter dendrites remain in pars lateralis. In total, the substantia nigra appears to have a layered organization: the superior layer is the cellular pars compacta, the second is the dorso-medial area of pars reticulata where both pars compacta and pars reticulata dendrites run rostro-caudally and dorso-ventrally and the third layer is the peripeduncular region where dendrites from all areas run parallel to the crus cerebri.


Microcapsules on Streptococcus mutans serotypes by electron microscopy. Extracellular microcapsules have been demonstrated on cells of most serotypes of Streptococcus mutans by electron microscopy, using bacterial strains of the various serotypes and peroxidase labeled or unlabeled immune serum. A correlation was noted between the amount of capsular substance on the strains of S mutans examined and degree of antigenicity as expressed by the indirect fluorescent antibody (FA) title. A serotype d strain was shown to lose both antigenicity as determined by the FA reaction and capsular material as seen by electron microscopy with repeated in vitro passage. When 10% unheated rabbit serum was added to the medium, antigenicity and capsular material were restored.


Studies of perennial ragweed immunotherapy. Perennial ragweed immunotherapy was studied in 24 patients with ragweed pollenosis. Cellular responsiveness was determined by measuring the cellular reactivity and sensitivity to ragweed antigen E (RW-AgE) by RW-AgE-induced leukocyte histamine release. Serum blocking antibody content was determined by measuring the serum RW-AgE binding capacity by ammonium sulfate coprecipitation of bound RW-AgE. Specific IgE (anti-RW-AgE) concentration was determined by polystyrene tube radioimmunoassay. Cellular responsiveness decreased with continuing immunotherapy, as did the specific IgE concentrations. The serum RW-AgE binding capacity, in contrast, increased as treatment continued. The absence of demonstrable correlations between RW-AgE binding capacity and cellular reactivity, cellular sensitivity, and specific IgE concentrations contrasted impressively with the demonstration of multiple significant correlations between the change in the RW-AgE binding capacity and the other parameters studied. The degree of increase in RW-AgE binding capacity correlated significantly with the degree of decrease in both the specific IgE concentration (p is less than 0.04) and the cellular sensitivity to RW-AgE (p is less than 0.003). these findings suggest that the active process of blocking antibody production, rather than the passive presence of blocking antibody, is related to the process which decreases the specific IgE concentration and the degree of cellular responsiveness and therefore results in clinical improvement.


Automated histamine analysis for in vitro allergy testing. II. Correlation of skin test results with in vitro whole blood histamine release in 82 patients. The recently developed sensitive, automated histamine assay system was applied for in vitro allergy testing. The simplified method for histamine release from whole heparinized blood was used. Aliquots of blood and allergen were incubated for one hour at 37 degrees C, and each supernatant was then analyzed for histamine release. Nine common pollen and environmental allergens were used at three 10-fold dilutions for in vitro testing with the use of 20 ml of blood. Intradermal skin tests were correlated with the whole blood histamine release in 82 patients who had received no immunotherapy. A scoring system for the histamine results was developed to take into consideration the results with multiple allergen concentrations. When the skin test was strongly positive (greater than or equal to 3 + at 100 protein nitrogen units [PNU]/ml), the whole blood histamine release was positive in 89% of the tests. In contrast, when the skin test was negative ( less than 1 + at 100 PNU/ml), the histamine release was also negative in 99.8% of the cases. When the skin test was 1 +, the histamine release from whole blood was positive in 6% of the tests; and when the skin test was 2+, the whole blood results were positive in 32%. The accuracy, precision, and sensitivity of the automated histamine assay allow its application for the clinical study of allergic patients.