An adrenergic component of the nervous apparatus of the aortic reflexogenic zone. The structural organization of the adrenergic (sympathetic) component of the autonomic innervation of the depressor zone of the cat aortic arch was studied by luminescence microscopy of the catecholamines. A terminal adrenergic plexus, branching extensively in the connective-tissue basis of the depressor area of the aortic arch, was discovered. The participation of vessels supplying blood to the depressor area was established. Adrenergic neurons were found in the territory of the depressor zone of the aortic arch. It is postulated that the adrenergic component of the depressor zone of the aortic arch participates in the peripheral mechanism of the regulatory effects of the sympathetic nervous system on the baroreceptor apparatus.


Carcinoma of the pancreas and periampullary region: a clinical study in a district general hospital. One hundred patients with pancreatic and periampullary carcinoma have been reviewed. Diabetes mellitus may be an early presentation. Painless jaundice has a relatively good prognosis. Fifteen patients have had curative procedures with a 31 per cent operative mortality. Mortality is influenced by age and serum bilirubin levels. Where palliative procedures are undertaken, this should include biliary diversion and gastro-enterostomy.


Change in a myelin-associated glycoprotein in rat brain during development: metabolic aspects. The higher apparent molecular weight of the newly synthesized glycoprotein in immature rat myelin in comparison with that in mature myelin, which was originally demonstrated with radioactive fucose, was also shown by double labeling experiments with radioactive glucosamine or N-acetylmannosamine. In addition, periodic acid-Schiff staining of gels, on which the glycoproteins of mature and immature myelin were electrophoresed separately or mixed together, revealed that the stained glycoprotein in immature myelin had a higher apparent molecular weight. Age studies with radioactive fucose showed that the greatest difference in the apparent molecular weight of the newly synthesized glycoprotein occurred at 12 days. The magnitude of the difference decreased gradually with age until 25 days when the glycoprotein electrophoresed to the same position as that in more mature myelin. Long-term experiments, in which 12- or 13-day-old rats were injected with radioactive fucose and allowed to survive for varying periods of time before myelin isolation, also revealed a gradual decrease with age in the extent of the shift of the glycoprotein toward a higher molecular weight. The possible significance of the developmental change in the glycoprotein for the process of myelin formation is discussed.


Slow axonal transport or proteins; blockade by interruption of contact between cell body and axon. The influence of ligation and colchicine treatment on the axonal transport of slowly migrating [3H]leucine-labelled proteins was studied in the vagus nerve of the rabbit. Two days after [3H]leucine labelling of the dorsal motor nucleus of the vagus nerve, ligation or local application of 60 mM colchicine immediately blocked the further progression of slowly migrating proteins distal to the site of treatment. Application of 50-100 mug colchicine to the nerve cell bodies 2 days after labelling blocked the transport of slowly migrating proteins within the next 24 h. It is suggested that contact between nerve cell body and the axon is necessary for the maintenance of the slow transport of proteins in these nerves.


Transport of choline acetyltransferase and acetylcholinesterase in the central stump and isolated segments of a peripheral nerve. Axonal transport of choline acetyltransferase (ChAc, E.E.:2.3.1.6) and acetyl cholinesterase (AChE, E.C.:3.1.1.7) was studied in the peroneal fascicles of rabbit sciatic nerves. The accumulation of ChAc in the central nerve stump proceeded 5 times more slowly than that of AChE and occurred at a distanct of 2-4 mm proximally from the end, whereas AChE accumulated in the last 2 mm of the stump. In double-ligated segments of the nerve in situ the activity of ChAc decreased at the proximal and increased at the distal end; the activity of AChE rose at both ends, The increase of ChAc activity did not cease until 22 h, whereas that of AChE stopped before 10 h. The intensity of ChAc transport is considerably diminished in the part of axon separated from the nerve cell body. Differences between the behavior of ChAc and AChE are interpreted by the assumption that the axonal transport of ChAc is slow, unidirectional, concerns all of the enzyme in the nerve, and that most of the transported enzyme is not associated with intraaxonal organelles. In contrast to ChAc, the transport of AChE is fast, bidirectional, and concerns a minor proportion of enzyme in the nerve; the transported enzyme is associated with organelles. The rate of proximodistal transport of ChAc is estimated at 4 mm/day (based on the assumption that 100% of the enzyme moves proximo-distally) and that of AChE at 480 mm/day (based on the extimate that 5% of enzyme moved proximo-distally in the present experiments).


Environmental factors affecting the degradation of Dyfonate by soil fungi. The ability of selected fungi to degrade the soil insecticide Dyfonate (O-ethyl S-phenyl ethylphosphonodithioate) into water-soluble, noninsecticidal metabolites was found to be dependent on the supply of nutrients, incubation time, temperature, pH, as well as other factors. With yeast extract as the carbon source (5 g/liter) and ammonium nitrate (1 g/liter) as the nitrogen source, both Rhizopus arrhizus and Penicillium notatum degraded the insecticide to a larger extent than with any other combination of nutrients used. With glucose as the carbon source, concentrations of ammonium nitrate above 5 g/liter inhibited the degradation of Dyfonate by R. arrhizus. Time-course studies on the metabolism of the insecticide indicated that Dyfonate was first absorbed by the fungal mycelium, where it was metabolized followed by the release of water-soluble, noninsecticidal, breakdown products into the culture media. The degradation appeared to involve the breakdown of Dyfonate into ethyl acetate soluble metabolites, such as ethylethoxyphosphonothioic acid, ethylethoxyphosphonic acid, methyl phenyl sulfoxide, and methyl phenyl sulfone. These compounds were then further degraded into water-soluble products. The optimum conditions for the degradation of the insecticide by R. arrhizus were observed at pH 6.0 to 7.0 and at 15-25 degrees C. Aged fungal mycelia were as active as mycelia in the logarithmic growth phase.


Effect of carbon source during growth on sensitivity of Pseudomonas fluorescens to actinomycin D. Sensitivity to actinomycin D(AD) varies in Pseudomonas fluorescens cells grown in glucose or succinate minimal salts medium. Growth is inhibited in succinate minimal medium by much lower concentrations of AD than in glucose minimal medium. Uptake of selected radioactive metabolites is inhibited by AD in cells incubated for 2 h in succinate medium containing AD but glucose-grown cells were not sensitive. EDTA treatment promotes increased sensitivity to AD in succinate-grown cells but does not alter sensitivity in glucose-grown cells. Succinate-grown cells bound 2-3 times as much 3H-AD as glucose-grown cells. Glucose-grown cells had much higher lipopolysaccharide levels in the envelope than succinate-grown cells. It is proposed that the lipopolysaccharide masks the binding sites and, therefore, is responsible for the difference in binding of AD by the glucose- and succinate-grown cells. The availability of the binding sites is also reflected in the sensitivity of the cells to the antibiotic.


Fine structure and distribution of extracellular polymer surrounding selected aerobic bacteria. The structure and distribution of extracellular polymer surrounding Bacillus circulans, Diplococcus (Streptococcus) pneumoniae, Streptococcus salivarius, Staphylococcus aureus, Klebsiella pneumoniae, Pseudomonas aeruginosa, Herella vaginacola (Acinetobacter calcoaceticus), and Agrobacterium tumefaciens were studied by electron microscopy. A modified ruthenium red staining procedure was used to examine the fine structure of capsule and slime. Freeze-etching and critical-point drying were used to examine the quantity of unaltered exocellular material. Comparative data demonstrate that fibrillar extracellular polymer surrounding B. circulans, D. pneumoniae, and K. pneumoniae is capsule (cell wall attached) which is characteristic of the producing organism. Capsular polymer generally appeared fibrillar, although globular polymer consisted of capsular subunits bound to S. salivarius and H. vaginacola. Exocellular slime was present about S. aureus, P. aeruginosa, and A. tumefaciens.


Prophylaxis of varicella in children with neoplastic disease: comparative results with zoster immune plasma and gamma globulin. The incidence and severity of varicella following a close family contact were evaluated in children with neoplastic diseases who received prophylaxis either with commerical gamma globulin or with zoster immune plasma, as compared to patients who did not receive any prophylaxis. In the untreated group, all 14 patients developed varicella, complicated by 1 case of encephalitis and 2 cases of fatal pneumonia. In the group of 17 patients who received 0.6-1.2 ml/kg body weight of gamma globulin, 16 developed varicella, complicated by pneumonia in 2 cases, with 1 death. In the third group of 27 patients who received 10 ml/kg body weight of zoster immune plasma (ZIP), obtained from healthy adults convalescing from herpes zoster, there were only 8 cases of varicella, all very mild. Thus, prophylaxis with ZIP significantly reduced the incidence of clinical varicella (p less than 0.01) and attenuated the severity of its course.


A clinical evaluation of indium-111 bleomycin as a tumor-imaging agent. In an evaluation of indium-111-bleomycin as a tumor-imaging agent, 357 whole-body tumor scans were performed in 293 patients. Of 246 studies performed in patients with a variety of active solid tumors, 218 (89%) were true-positive studies and 28 (11%) were false-negative. Of 69 scans in patients thought to be free of tumor after therapy, 32 (46%) were false-positive studies and 37 (54%) were true-negative. The true-positive rates by major tumor type were: adenocarcinoma of gastrointestinal tract origin (95%), lymphoma (88%), melanoma (87%), sarcomas (82%), lung (77%), breast (77%), childhood tumors (71%), gynecologic tumors (70%), and genitourinary tumors (68%). Soft tissue and lymphatic sites of tumor, both above and below the diaphragm, were easily visualized, whereas hepatic and bone marrow sites of involvement were less easily discerned. False-positive uptake with 111In-bleomycin was noted in lungs (6%), gut (3%), mediastinum (2%), normal breast tissue (0.8%), and in occasional inflammatory lesions. In 19 patients with multiple myeloma or leukemia, a pattern of diminished bone marrow uptake associated with abnormal accumulation of 111In-bleomycin in extramedullary sites of involvement was the rule. In another 23 patients in whom scans were performed because an occult tumor was suspected, scanning did not lead to specific diagnosis of tumor in a single instance. We conclude that 111In-bleomycin is a safe, effective, and useful new tumor-imaging agent in the initial staging and followup of patients with a variety of solid tumors. Significant advantages of this agent over other currently available radiopharmaceuticals include: A) a broader spectrum of tumors taking up the radio-pharmaceutical, and B) generally better delineation of abdominal and pelvic disease due to lack of interference from gut uptake.


Morphological basis for the cytolytic effect of vinblastine and vincristine on cultured human leukemic lymphoblasts. Exposure of human leukemic lymphoblasts in suspension cultures to low concentrations of vinblastine and vincristine results in alterations in cell shape and leads to the formation and release of a large number of membrane-lined vesicles from the cytoplasm. Separation of these vesicles from peripheral cytoplasm is effected through alignment and fusion of small vacoules. Similar vesicle formation is seen neither in fibroblasts exposed to vinblastine nor in lymphoblasts exposed to bleomycin or adriamycin. Possible relation of this phenomenon to vinblastine- and vincristine-induced cytotoxicity, spherocytosis, and thrombocytosis is discussed.


Identification of lymphoid cells in cultured of murine leukocytes and thymus. Several culture conditions and media were studied in an effort to establish long-term cultures of murine lymphoid cells from blood and thymus. Cultures vessels included small glass bottles and rubber-stoppered tubes. Media such as Roswell Park Memorial Institute 1640,  1700, 1701, 1715, GEM 1717, NCTC, fetal calf and horse serum supplements, and conditioned medium were tried. Lymphoid cells in mouse leukocyte cultures survived as long as eight months before dying out. However, lymphoid cells in thymus cell cultures, strarted and maintained with GEM 1717 medium with 20% fetal calf serum supplementation, gave rise to cell lines that continued to yield subcultures for more than 2 years. Mcroscopic examination of thymus cell subcultures revealed lymphoid and thymic epighelioid cells. Tumorigenicity studies of one cell line were negative. Chromosomal preparations of this cell line often contained near-normal karyotypes but were complicated by the presence of binucleated cells. Live cell fluorescent antibody assays for surface theta-antigen and immunoglobulin revealed immunoglobulin-negative cells possessing barely detectable theta determinants. Functional assays for thymus-derived lymphoid cell activity suggested that these cells were mitogen responsive and weakly reactive in one-way mixed lymphocyte culture. On the basis of this evidence it was sugguested that these cells represent a class of T-cells (thymus-derived lymphocytes) that have all but  lost theta antigen, possibly due to prolonged culture.


Pepstatin, an inhibitor of leukokinin formation and ascitic fluid accumulation. Ascites fluid accumulation accompanying a mastocytoma or L1210 murine tumor is significantly retarded following the i.p. or s.c. injection of moderate quantities of pepstatin, a known acid protease inhibitor. No effect on cell count was noted by pepstatin treatment. The probable mechanism by which pepstatin acts is by inhigiting the enzymatic formation of chemical mediators known as leukokinins. These are pharmoacologically active peptiedes having potent  permeability characteristics previously described by this laboratory. Leukokinins are formed by cathepsin D-like enzymes present in the invading cells and in the ascites fluid acting on a protein substrate, leukokininogen. present in the ascites fluid. Pestatin inhibits the action of these leukokinin-forming enzymes invitro but has no effect on kallikreins (bradykinin-forming enzymes) in vitro. Human ascites fluid from a patient with ovarian carcioma was found to have a paepstatin-inhibited, leukokinin-generating system, as does the mouse. A 'chemical mediator' theory is proposed for ascites fromation which broadens the previously held theory of lymphatic blockage (Holm-Nielsen) and may explain the recent findings of Hirabayashi and Graham of increased plasma-ascites exchange in peritoneal carcionmatosis. Pepstatin inhibition of chemical mediator formation may represent a new therapeutic approach to ascites fluid accumulation in neoplastic disease.


The morphology of murine oncornaviruses following different methods of preparation for electron microscopy. The effect of different preparative procedures for electron microscopy on the size and shape of murine oncornaviruses has been studied. With conventional negative staining procedures using neutral sodium phosphotungstate, both murine mammary tumor virus and murine leukemia virus appeared in head-and-tail forms, with a peak head diameter of 122 and 130 nm, respectively. Negative staining with uranyl accetate gave round virions with peak diameters of 148 and 130 nm. Prefixed virus was round with peak diameters of 141 and 130 nm, respectively, in phosphotungstate, and 148 and 117 nm, respectively, in uranyl acetate. With thin sections, the peak diameters were 143 and 123 nm. The preservation of  the spherical shape of the virus was obtained by glutaraldehyde fixation dehydration in alcholic solutions of uranyl acetate, and critical point drying. Under these conditions the viruses had peak diameters of 99 and 82 nm, respectively. The size of murine mammary tumor virus has always  been found to be larger than murine leukemia virus in all preparations except for negative staining with neutral sodium phosphotungstate. Shadowing of the virion preparations revealed considerable flattening of the particles in all cases except for critical point drying. Negatively stained preparations did not cast any shadow, and thus thethickness of the particles could not be evaluated. Virus can be reversibly converted from spherical to head-and-tail forms by altering osmotic strength. Under most of the conditions used, murine mammary tumor virus gave a bimodal size distribution with significant numbers of particles that were smaller than the major virus size.


Biochemical characterization of putative subviral particulates from human malignant breast tumors. Particulates with the properties of cores and/or ribonucleoproteins of RNA tumor viruses have been isolated from Sterox-SL-treated fractions of murine and human mammary adenocarcinomas. These particulates have an RNA-directed DNA polymerase, a 60 to 70 S RNA, and a density of 1.26 g/ml or greater in sucrose equilibrium density gradients. Their uniquely higher densities lead to banding in regions comparatively free of cellular contaminants. These circumstances minimize some of the technical complications of performing the simultaneous detection assay in the presence of cell debris.


Tissue-specific DNA-protein complexes during azo dye hepatocarcinogenesis. The immunological tissue specificity could be transferred from one chromatin preparation to another by reconstituting this protein fraction to the DNA and the remaining chromatin components."The immunological tissue specificity could be transferred from one chromatin preparation to another by reconstituting this protein fraction this protein fraction to the DNA and the remaining chromatin components.


Serum alpha-fetoprotein in patients with neoplasms of the gastrointestinal tract. Serum alpha-fetoprotein levels were measured by a sensitive double-antibody radioimmunoassay in 580 patients with a variety of malignant and nonmalignant gastrointestinal diseases to determine the incidence of levels elevated above 40 ng/ml. Over 200 normal control subjects have all had levels below 40 ng/ml. Fifteen % of 95 patients with gastric carcinoma, 3 percent of 191 patients with colorectal carcinoma, 24 percent of 45 patients with pancreatic carcinoma, 25 percent of 8 patients with biliary tract carcinoma, and 70 percent of 73 patients with hepatocellular carcinoma had elevated serum alpha-fetoprotein. None of 14 patients with esophageal or small bowel carcinoma had elevated levels. In contrast, 1 percent of 154 patients with nonmalignant, nonhepatic gastrointestinal disease had elevations of serum alpha-fetoprotein. Alpha-Fetoprotein appears to be a potential marker for tumor activity in some patients with certain gastrointestinal cancers.


Detection of latent function in acutely ischemic myocardium in the dog: comparison of pharmacologic inotropic stimulation and postextrasystolic potentiation. In poorly perfused myocardium with resultant ischemic dysfunction, augmentation of contractility can, under certain conditions, be used to detect viable but ordinarily noncontracting muscle. Two methods of inotropic augmentation, pharmacologic inotropic stimulation and postextrasystolic potentiation (PESP), were studied in acutely ischemic canine myocardium with controlled coronary blood flow. A caliper length gauge to record segmental shortening and left ventricle pressure was used to construct pressure-length loops. Acute regional ischemia depressed segmental function: early segmental shortening decreased (-20 plus or minus 0.02% [SE]) and frequent dyskinesia occurred. Restoring coronary blood flow corrected segmental shortening to control levels. During acute regional ischemia, PESP consistently augmented segmental function (+49 plus or minus 0.03%) and abolished dyskinesia. Pharmacologic inotropic stimulation with isoproterenol or calcium administered into the coronary arteries did not produce a comparable improvement in segmental function (+9 plus or minus 0.05%). Although early shortening markedly increased with pharmacologic stimulation, there was no consistent change in total shortening, and the area of the pressure-length loop decreased. Due to late dyskinesia, there was a decrease in injection shortening. Systemically administered pharmacologic agents accentuated early dyskinesia but caused no consistent change in total shortening. Unlike PESP, pharmacologic agents either worsened segmental function or caused responses that were minimum and inconsistent; such responses clearly cannot be used to identify viable ischemic myocardium.


The heart as a target organ in systemic allergic reactions: comparison of cardiac analphylaxis in vivo and in vitro. The purpose of this investigation was to define and quantitatively evaluate cardiac anaphylaxis in vivo. Guinea pigs, passively sensitized with graded amounts of rabbit antipenicilloyl antibody, were anesthetized, ventilated, and challenged intravenously with a constant amount of antigen (anaphylaxis in vivo). In other experiments, guinea pig hearts were excised, perfused in a Langendorff apparatus, and challenged (analphylaxis in vitro). During in vivo anaphylaxis, sinus rate increased 10-30 beats/min, conduction arrhythmias occurred in 15 of 22 experiments, and ventricular fibrillation was seen in 8 of 22 experiments. Tachycardia and arrhythmias began approximately 20 seconds after antigen administration and were accompanied, but not preceded, by respiratory and pressor changes. During in vitro anaphylaxis, sinus rate increased 70-110 beats/min, coronary flow rate decreased 2-22%, conduction arrhythmias occurred in 21 of 31 experiments, and ventricular ectopic activity was seen in 13 of 31 experiments. Tachycardia and arrhythmias began approximately 15 seconds after antigen administration. Sinus tachycardia, atrioventricular conduction block, increased ventricular automaticity, and histamine release were characteristic features of cardiac anaphylaxis in vivo and in vitro. Both in vivo and in vitro, the intensity of the cardiac reaction depended on the amount of antibody used in passive sensitization. Our results clearly indicate that the heart reacts as a target organ in systemic anaphylaxis of the guinea pig.


Three variants of concealed bigeminy.

Three variants of concealed bigeminy. Long electrocardiographic strips were analyzed from five patients who exhibited periods of typical "concealed bigeminy," i. e., recurrent unifocal extrasystoles which were separated from one another by odd numbers of normally conducted sinus beats.  However, in each of these patients, there were periods in which one of three different variants of concealed bigeminy was observed.  Three patients displayed an "even number" variant; i. e., there were large numbers of consecutive extrasystoles which were separated exclusively or preponderantly by even rather than by odd numbers of sinus beats.  One other patient exhibited an "interpolated extrasystole" variant: those interectopic intervals which were initiated by an interpolated extrasystole contained an even number of sinus beats, whereas all other interectopic intervals contained an odd number.  In the fifth patient, the distribution of the numbers of sinus beats separating extrasystoles was such as to suggest a periodic fluctuation between the classical forms of concealed bigeminy and concealed trigeminy; i. e., a "combined bigeminy and trigeminy" variant.


De subitaneis mortibus. XI. Young girl with palpitations. A seventeen-year-old girl in apparent good health was found dead in bed one morning.  Two clinical features of note were multiple premature beats present in her electrocardiograms for several years, and a tall thin habitus without stigmata of Marfan's syndrome.  At necropsy examination the heart was grossly normal.  At its margin abutting the central fibrous body the atrioventricular (A-V) node exhibited numerous fronds and outcroppings, some forming loop connections from one part of A-V node to another.  Groups of A-V nodal cells detached from the A-V node were also found connected to the crest of the interventricular septum.  There were large connections directly from the His bundle to the crest of the interventricular septum (Mahaim fibers).  The A-V node artery was moderately narrowed but patent.  The possible significance of these findings in the pathogenesis of re-entrant arrhythmias or ectopic automaticity is discussed.


Diagnostic value of routine liver tests. In 131 patients on a medical service and 97 patients on a surgical service, in whom a diagnosis of hepatobiliary disease was verified in the hospital, the diagnostic value of routine liver tests performed soon after admission was evaluated by stepwise discriminant analysis. By measurements of alanine aminotransferase, alkaline phosphatases, gamma globulin, prothrombin time, bilirubin, and albumin, half of the medical patients were correctly classified into one of seven diagnostic categories. Aminotransferase contributed most to the classification, being twice as effective as random allocation. Decreasing the number of diagnostic categories to three (hepatitis, fatty liver, and chronic liver disease) increased the frequency of correct allocation to 80%. The allocation of all the patients to seven medical and four surgical diagnostic categories by means of four tests (aminotransferase, alkaline phosphatases, prothrombin time, and bilirubin) was significantly improved by each step with a misclassification rate of 55% when all tests were used. A reduction of the diagnostic groups to five (hepatitis, fatty liver, chronic liver disease, duct obstruction and tumor) increased the frequency of correct allocation to 63%. The analysis demonstrates the limited diagnostic effectiveness of routine liver tests when used alone. The absolute discrimination values depend on the a priori frequencies of the diagnostic groups investigated, and therefore may vary from time to time and from place to place.


Microanalytical system for determination of picogram quantities of metals in metalloenzymes, as illustrated with zinc-containing enzymes. Microwave-induced emission spectrometry combined with gel-exclusion chromatography provides a microanalytical system capable of precisely measuring 10 minus 10 to 10 minus 13 g of metal in microgram amounts of enzyme.  Such sensitivity greatly exceeds that of other, more conventional methods.  Metal quenching agents and low-molecular-weight protein contaminants were removed from the enzyme by Sephadex G-100 chromatography in microbore columns (0.03 times 25 cm).  Droplet fractions were analyzed for zinc by the present method, for enzyme activity, and for protein content.  With this analytical system we could demonstrate that stoichiometric amounts of zinc are present in the RNA-dependent DNA polymerase, the reverse transcriptase, from wooly monkey type C RNA tumor virus.  The precision of the method for zinc was demonstrated by the coefficient of variation of 4.4 percent for 10 mug of zinc per liter.  Validity and accuracy of the method were established by determining zinc in a series of zinc metalloenzymes of known metal content and stoichiometry.


Time course of development of supersensitivity to topical acetylcholine in partially isolated cortex. In alert monkeys the time course for development of supersensitivity to topical acetylcholine in partially isolated frontal cerebral cortex was determined. Thresholds for paroxysmal discharge fell progressively and markedly during 3 weeks, further in 5 and somewhat more after 6 months. ACh supersensitivity was demonstrated in chronic "isolated" occipital cortex. Epileptiform discharges were recorded selectively from chronic partially isolated frontal cortex on peripheral nerve stimulation and these spread, causing a clinical convulsive siezure when the open end of the isolation extended into the precentral gyrus. The basic mechanisms responsible for the supersensitivity are unknown but evidence presented and much in the pertinent literature is in keeping with the hypothesis that partial isolation of cortical cells, i.e., denervation, deafferentation, or disuse may be important. It is suggrested that peripheral nerve stimulation, like arousal, may cause an outflow of ACh on the normal brain surface and over the open end of a partially isolated area, which, especially, in the presence of a diminished cholinesterase activity (in partially isolated cortex), could act like topical ACh, cause a DC shift and an epileptiform discharge.


Effect of hippocampectomy on sleep patterns in cats. The study was planned to see if the hippocampus has an influence on fast wave sleep (FWS) as well as on slow wave sleep (SWS). From 8 male cats EEG, EMG and EOG were recorded for 24 h, first under normal conditions, secondly after cortical damage to the dorsal marginal portion of posterior ectosylvian gyrus, and thirdly following hippocampectomy done through the cortical damage. From the records, SWS, FWS and the sleep state (defined as a sequence of SWS or SWS-FWS phases between two successive waking states) were measured in terms of their occurrence, the mean duration and the total time they occupied in the day, night and 24 h. In addition, sleep sequences were classified according to the number of constituent sleep phases. Cortical damage did not affect SWS, FWS, or sleep state with regard to their occurrence, the mean duration, and the total time they occupied in 24 h. Nor did it affect the proportion of short and long sequences. The circadian variation of sleep was clearly retained. Hippocampectomy significantly reduced the total time occupied by sleep state, SWS and FWS, increased the occurrence of sleep state and SWS phase against decreased incidence of FWS phase, and reduced the mean duration of sleep state and SWS phase. Hippocampectomy also significantly increased the occurrence of sleep sequences with only one SWS phase at the cost of sequences with alternating SWS and FWS phases. Following hippocampectomy, the circadian variation of sleep was not only retained, but actually exagerated. The hippocampus in inferred to facilitate the FWS as well as the SWS phase of sleep.


Synchronization of human motor units: possible roles of exercise and supraspinal reflexes. Some normal human subjects show definite synchronization of the motor units in hand muscles (i.e., the impulses from two or more motor units coincide in time more frequently than expected for independent random processes). Subjects who show synchronization tend to use their hands to exert large, brief forces, either in their work (e.g., manual laborers) or recreational activities (e.g., weightlifters). In this study all seven weightlifters examined showed a significant degree of synchronization. Furthermore, after 6 weeks of using the first dorsal interosseus muscle of the hand to exert maximal, voluntary contractions, the level of synchronization increased substantially in four control subjects, and the average level became significantly different from zero. Weightlifters also showed greater late reflex responses than control subjects, but no significant difference in earlier spinal reflexes. Two late reflex waves are described which probably involve fast pathways to and from motor cortes. We suggest that supraspinal connections from motor cortex directly to spinal motoneurons may be enhanced as a result of training to the point where they produce a significant synchronization of motor units during steady, voluntary contractions.


The late positive component (P300) and information processing in sentences. Averaged visual evoked potentials to sequentially flashed words comprising a sentence were recorded from vertex and left and right temporoparietal electrodes in 8 right-handed subjects. In condition 1 the sentence took the form: The -eel is on the shoe, in which the first grapheme was omitted from the second word, so that the subject did not know the meaning of the second word until he viewed the last word. In condition 2, the sentence took the form: The heel is on the shoe, in which the second word was given and the last word provided no further information. P300 latency to words which delivered information (last word of condition 1, second word of condition 2) were significantly longer than P300 latency to any of the other words in the sentence, as well as to the same position word in the other condition. Comparisons of P300 latencies to redundant words (the, is, on) within and between conditions showed no significant differences. P300 amplitude to the last word was significantly larger than P300 amplitude to any of the other words within the sentence, even in condition 2 where the second word delivered information. The major effect of information delivery was on P300 latency, while "syntactic closure" had its major effect on P300 amplitude. The fact that evoked potentials to all words had P300 components was attributed to the engagement of the P300 system whenever task-related language stimuli are used.


Cholinergic mechanisms involved in cortical activation during arousal. Spontaneous and evoked cortical electrical activity of the post-central gyrus was studied in 5 unparalyzed, unanesthetized monkeys during slow-wave sleep (SWS) and wakefulness (W), before and after the administration of a muscarinic anticholinergic agents, hyoscine. It was found that hyoscine reproduces the effect of SWS: (1) on spontaneous EEG activity and single unit discharges of the somatosensory cortes without any concomitant behavioral sleep; and (2) on surface evoked potentials and reactivity of cortical neurons to peripheral electrical stimulation. Short episodes of EEG desynchronization were noted in the alert state after hyoscine suggesting the existence of a phasic activating system unblocked by hyoscine. These observations support the concept of an EEG-behavior dissociation produced by muscarinic anticholinergic agents and extend this concept to single neuron activity. These observations also lead to the conclusion that tonic EEG activation during arousal may be cholinergic at the cortical level even though other neurotransmitters may be involved.


Motor potentials and the timing of muscular activity. The purpose of this study was to determine whether dominant components of the motor potential recorded from the scalp, were coincident with the rise and fall of muscular activity in single movements and return movements in serial flexion and extension of the elbow. Electroencephalographic recordings obtained from two right-handed subjects through scalp electrodes were averaged by computer and related to EMG, force and displacement of the limb. In the single movements, coincident events in the motor potential indicated the rise and fall of EMG and force. Events in the motor potential of the return movement coincided with the rise of EMG and of force in the first phase, and the rise of EMG in the second phase. Other time relationships were not clear. Relationships between events were clearer in one subject than in the other. This could have been caused by different characteristics in the performance of the task or by problems related to the location of scalp electrodes. The general waveform of the motor potentials and the EMG traces seemed to be related in a few samples of the single movement task, suggesting that other factors than time may also be related between the motor potential and the EMG.


Intracortical transmission of specific evoked activity across the cerebral cortex. The intracortical transmission of abnormally generated activity, like spreading depression and the direct cortical response, across the cortex is well known. In the experiments described here, the tangential transmission of a more normal type of cortical activity, the short latency response evoked by contralateral forepaw stimulation, was investigated. No evidence for tangential transmission was found from surface recordings. Various methods, such as passing polarizing currents, intravenous injection and topical application of strychnine, were used to alter the state of cortical excitation in order to induce a change in tangential transmission; none was found. The boundaries of the contralateral forepaw receiving area (CRA) appeared to be rigid, since the responses could not be induced beyond the perimeter by these manipulations of the cortex. Evoked responses in the centre of the CRA were shown to be unaffected by the excitation state of the cortex 2 mm or more away. The data support the hypothesis that the neural activity represented by the short latency surface evoked response is not transmitted laterally across the cortex and does not appear to be influenced by activity, beyond a range of about 2 mm or possibly less. The effectiveness of the methods used to alter cortical excitation, and to detect a spread of the evoked activity are discussed.


Eye movement related activity in the visual cortex of dark-reared kittens. Kittens reared in total darkness from birth were found to be behaviorally blind, when tested at the age of 15-20 weeks. Visual cortex EEGs were recorded with transcortical electrodes. During waking, potentials related to saccadic eye movements (EMP's) were present in the dark, though they were depressed in amplitude in the light. During paradoxical sleep, rapid eye movements, isolated or in bursts, were present at a normal rate, as were occipital waves related to ponto-geniculo-occipital activity. It is concluded that EMPs during waking and phasic bursts during paradoxical sleep represent central "built in" events uninfluenced by visual experience.


[Polygraphic study of sleep in young chickens at hatching, evolution at third and fourth days]. A polygraphic study of the states of vigilance in the young chicken was performed in the first week after hatching. At hatching, three stages of vigilance were present and their polygraphic characteristics were similar to those of the adult. Quantitative data showed a relative immaturity of the young chicken since percentages of paradoxical sleep and slow wave sleep were high in the first 8 h after hatching (PS/TS equals 16.5%). These values diminished rapidly and reached the adult values observed on the third day (PS/TS equals 6.4%). Our experimental conditions showed the presence of a nycthemeral rhythm of the states of vigilance by the third day.


[A telemetric study of central rhythms in children]. Data concerning central (mu) rhythms in children are reported, these rhythms being studied by means of prolonged telemetric recording which makes it possible to study various experimental situations. Several different patterns of mu rhythm were observed in every subject, depending on the experimental situation. The level and the quality of attention on the one hand, immobility on the other, were found to be the two main factors which facilitated the occurrence or the maintenance of mu rhythms. The induction of movement, especially of the hand, was the major cause of blocking pre-existing mu rhythms, but attention alone was also able to provoke the same reaction. These data were compared with those obtained in animals by direct recording from the cortical somaesthetic areas. The respective roles of vigilance, attention and movement are discussed.


[Influence of the electrical properties of electrodes (platinum wire) on the myoelectric signal]. The use, in electromyography, of platinum wire electrodes required a knowledge of the electrical characteristics of these electrodes in the frequency range of the myoelectric signal. For this purpose the impedance of these electrodes was measured by the "cercle pointe" method (Gougerot 1951) in conditions similar to those in use. These electrodes behave like the majority of metal electrodes. Their impedance in particular can be represented by a polarization capacity shunted by an ohmic resistance, in series with a resistance representing the electrolyte resistance. It is shown that, depending on the electrode area and the way they are used, these electrodes can detect the myoelectric signals more or less selectively.


A chronic electrode implantation technique for sub-mammalian vertebrates. A new microelectrode implantation technique is described for recording electrical activity in the brains of awake, unrestrained small animals. The technique employs a miniature heating element and wax seal to enable movement of the microelectrode. Preliminary results in toads (Bufo americanus) are presented as an example of the technique.


[Conduction velocity of Ia sensory fibers in human measured by H-reflex method]. The conduction velocity in Ia sensory fibres of the peroneal nerve was measured in 20 normal adult subjects. The method consisted of stimulating the nerve at two different sites, separated by a known distance, and measuring the latency differences of the monosynaptic reflex responses.


Single unit activity in the lateral amygdala of the cat during sleep and waking. Neuronal activity of 58 cells in the lateral nucleus of the amygdala of freely moving cats was studied across stages of sleep-wakefulness. In general, these units were very slow firing in all states (mean discharge rate less than 1 spike/sec) with the highest rates typically observed during slow wave sleep and paradoxical sleep. Most units tended to show some bursting activity during all states. The bursting during paradoxical sleep was not temporally correlated with the phasic activity characteristic of that state. These results corroborate and extend previous studies of the basal amygdaloid nuclei.


An application of long-term frequency analysis in measuring drug-specific alterations in the EEG of the cat. A method is described for the quantitative measurement of "drug-specific" effects on the EEG of the cat. These effects are dose-related and are independent of the normal sources of EEG variation associated with the sleep-waking cycle. Drug-induced changes are expressed as characteristic alterations in frequency spectra and the time courses of these effects are followed for 5 h following administration of the test compounds. Atropine sulfate (0.5 and 2.0 mg/kg) and physostigmine salicylate (0.05 and 0.20 mg/kg) were administered to three unanesthetized and unrestrained cats and a broad-band frequency analysis was performed on the spontaneous brain electrical activity recorded from the prepyriform cortex, ventral hippocampus, lateral geniculate nucleus and the midbrain reticular formation. The resulting data were used as input to discriminant and canonical statistical analysis programs employed to abstract "drug-specific" patterns of frequency change. It was found that both atropine and physostigmine produce alterations in EEG frequency spectra which are clearly distinct from those patterns associated with the sleep-waking cycle and thus neither compound results in what has been characterized as an "EEG-behavioral dissociation".


Spinal cord potentials evoked by peripheral nerve stimulation. Lumbar, thoracic, and cervical spinal cord responses evoked by sciatic nerve stimulation were measured in 20 cats at the skin level and directly from the dorsal surface of the cord. Computer averaging techniques were used for both skin level recordings and cord surface recordings at all levels. Recordings made directly from the cord surface at lumbar levels were large and were characterized by a large, initial, negative transient, followed by a more complicated and variable waveform. As recording electrodes were moved rostrally, the initial large spike decreased in amplitude and the duration and latency of the response increased. At cervical levels were large and were characterized by a large, initial, negative transient, followed by a more complicated and variable waveform. As recording electrodes were moved rostrally, the initial large spike decreased in amplitude and the duration and latency of the response increased. At cervical levels the responses were polyphasic, of long duration and small amplitude. Deafferentation by posterior rhizotomy of all lumbar and sacral roots ipsilateral to sciatic nerve stimulation abolished the response at all levels. Thoracic cord section also abolished the response over the cervical cord. Skin level recordings were of shorter latency than direct recordings, especially at cervical levels. Response configurations were similar for both recording techniques at lumbar levels, but had different waveforms at both thoracic and cervical levels. Clinical implications of the techniques and possible explanations of the waveforms recorded are discussed.


Effects of procaine hydrochloride, diazepam, and diphenylhydantoin on seizure development in cortical and subcortical structures in rats. Procaine HCl and diphenylhydantoin (DPH) increased the duration and propagation of epileptiform afterdischarges (ADs) produced by electrical stimulation of the amygdala in rats. Procaine and DPH also increased the rate of seizure development (kindling) produced by repeated stimulation of the amygdala. Procaine and to a limited extentDPH would themselves act as convulsants in well kindled subjects. Diazepam, on the other hand, retarded or blocked amygdaloid kindling. Diazepam trigered a high frequency (20-30 c/sec) rhtthm in the amygdala, hippocampus and preoptic area. None of these drugs had any significant effect on potentials evoked in secondary limbic sites by single electrical pulses applied to the amygdala. Also, none of these drugs had any effect on recruiting or post-tetanic potentiation (PTP) in secondary sites produced by amygdala stimulation and none of the drugs had any effect on amygdaloid AD thresholds. The effects of these drugs on the responses evoked by anterior neocortex stimulation were quite different. Diazepam had no effect on any of the characteristics of the discharge or convulsion even at twice the dose levels used for the amygdala group. Procaine and DPH, however, blocked not only the eonvulsion but the AD as well. Eighty percent of the procaine- and DPH-treated rats failed to respond with neocortical AD even at current levels as high as 2000 muA. The few cortically stimulated subjects that did respond with an AD showed a subcortical rather than a neocortical seizure response. DPH had no effect on recruiting or PTP of the transcallosal response. Both procaine and DPH produced a weak but significant increase in the amplitude of the transcallosal evoked potential, while diazepam produce a weak decrement in that response.


An injury-induced diffuse slow potential from brain. Three different slow potential (SP) changes resulting from focal brain injury are described. The first is an immediate, high amplitude (in excess of 25 mV)negative shift at the site of injury. The second is a biphasic negative-positive SP wave which spreads throughout the cortex ipsilateral to injury and is similar to spreading depression (SD). The third SP change, called here the injury-induced diffuse slow potential (IDSP)is a prolonged (lasting approx. 2h) negative shift occurring simultaneously in many brain areas, also in those far removed from the injured focus. The SD can be separated from IDSP by the size of focal injury; a 20 mu pucture of the parenchyma will trigger IDSP but not SD. An injury resulting from a larger puncture triggers both, SD and IDSP. IDSPcan not be induced by a re-entry of a previously damaged tissue. The magnitude of IDSP has anatomical specificity in that the largest amplitude occurrs in white as compared to gray of the cortex or of the caudate nucleus. Aso, the magnitude of the hypothalamic IDSP is larger when ipsilateral corpus callosum-commissural regions are injured. Electrical stimulation of the cortex in rats sufficiently strong to result in tonic-clonic convulsions triggers SD and IDSP; these two slow potential changes are similar to those induced by mechanical injury. A transpinnate electrical stimulus strong enough to elicit a grand-mal type of discharge results in a diffuse negative slow potential change similar to IDSP elicited by mechanical damage or direct cortical stimulation.


Human frequency-following responses to monaural and binaural stimuli. Frequency-following responses, with latencies circa 6 msec, were recorded from five normal-hearing human subjects to brief 500 c/sec tone bursts presented monaurally. The frequency-following responses appear as peaks occurring at 2 msec intervals superimposed on a slow wave (pedestal-like) component. Comparisons were made between the frequency-following responses evoked by binaural and monaural stimuli. The results show that the binaural responses may be interpreted as the sum of two monaural responses. It is concluded, therefore, that there are two independent populations of neurons, each capable of generating a frequency-following response is not a microphonic-like response but rather that the individual waves in the frequency-following response are evoked by the collective activity of phase-locked single units. Finally, on the basis of the distinctness of the individual waves in the frequency-following response, it is concluded that the neural generators of the response must be spatially compact.


Two varieties of long-latency positive waves evoked by unpredictable auditory stimuli in man. Two distinct late-positive components of the scalp-recorded auditory evoked potential were identified which differed in their latency, scalp topography and psychological correlates. The earlier component, called "P3a" (latency about 240 msec), was elicited by infrequent, unpredictable shifts of either intensity or frequency in a train of tone pips whether the subject was ignoring (reading a book) or attending to the tones (counting). The later component, called "P3a" (mean latency about 350 msec), occurred only when the subject was actively attending to the tones; it was evoked by the infrequent, unpredictable stimulus shifts, regardless of whether the subject was counting that stimulus or the more frequently occurring stimulus. Both of these distinct psychophysiological entities have previously been refered to as the "P3" or "P300" in the literature.


A syndrome of hypersomnia with automatic behavior. Five male patients, complaining of daytime sleep attacks, and hav ing a history of automatic behavior, were recorded continously by polygraph for 108 h under two different protocols: (1ents were given specific tests to perform during Phase 1. Six normal males were run as controls during Phase . At the end of the recordings, patients had measurements of homovanillic acid (HVA) and 5 Hydroxy-indol Acetic Acid (5HIAA) in CSF  before and after Probenecid testing. Patients appeared to present an abnormal sleep structure, with lack of Stage 3 and 4 NREM sleep, but had no significant increase in total sleep time during 24 h compared to normals. They also had repetitive very short periods of "micro-sleep" which account for their automatic behavior and were responsible for their great decrement in performance compared to normal controls. No CSF HVA and 5HIAA abnormality could be detected. This syndrome, which greatly impairs the social life and working ability of the patients, may be more related to an impairment of the "wakefulness" structures than to a dysfunction of the "sleep" structures.


EEG correlates of visual-motor practice in man. Special analysis of EEG signals was performed for 15 subjects engaged in three motor tasks of differing difficulty. A measure of average weighted coherence (C) was computed between the six possible combinations of four scalp areas: Oz, C3, C4 and Fz. In all subjects, regardless of task, scalp recordings over cortical areas known to have relatively dense fiber connections had significantly greater C values. However, the effects of task difficulty and practice were superimposed upon this basic pattern. Thus, the most difficult task (pursuit-rotor tracking) resulted in the highest coherence levels, while the least difficult task (visual tracking only of the pursuit-rotor disk) resulted in the lowest coherence levels. Practice, on the other hand, was associated with a significant decrease in overall level of coherence. This decrease is consistent with an interpretation of reduced task difficulty due to visual-motor learning. The results of the present study suggest that patterns of scalp EEG coherence may reflect some aspects of the underlying pattern of anatomical pathways, as well as the more dynamic properties of task difficulty and visual--motor practice.


Cortical seizures following cerebellar stimulation in primates. The effects of cerebellar stimulation were studied in monkeys with chronic alumina-cream epileptogenic foci in motor cortex. Low-frequency stimulation (5-15 c/sec) was ineffective in altering spontaneous cortical spiking. Clinical and electrographic seizures were elicited following high-frequency cerebellar stimulation (100 c/sec). This was a consistent finding following cerebellar stimulation, although no spontaneous seizures had been seen in these animals. These studies suggest the existence of facilitatory cerebellar mechanisms and indicate the need for further studies in chronic animal models.


The pattern of swallowing during sleep.

The pattern of swallowing during sleep. The pattern of swallowing during sleep was investigated by means of a study in 10 normal subjects. Sleep was staged by means of EEG, EOG and EMG, and swallowing was monitored by means of an external sensing device attached to the neck. It was found that during sleep, swallowing is episodic, with long swallow-free periods. Swallows occur almost exculusively in association with movement arousals which are most frequent during Stages REM, 1 and 2 of sleep.


Ontogeny of B-lymphocyte function with respect to the heterogeneity of antibody affinity-1,2. Neonatal liver or adult spleen was used as a source of B-lymphocytes in reconstituting lethally irradiated, syngeneic mice. Recipients were all given excess adult, syngeneic thymus cells and were immunized with dinitrophenylated bovine gamma globulin. The distribution of avidities of plaque-forming cells produced by immunized recipients of neonatal liver was highly restricted in comparison with animals reconstituted with adult spleen indicating a restriction of B-lymphocyte heterogeneity in the neonatal mouse.


Effects of aging on the differentiation and proliferation potentials of cells of the immune system. An attempt has been made here to show that the immune system can begin to decline in function shortly after an individual reaches maturity. The decline is due in part to changes in the environment of the cells but primarily to changes in the precursor cells of the system. This is reflected in their inability to proliferate and possibly differentiate efficiently. These findings show that the immune system can serve as an excellent model to study how aging can perturb the process of cells undergoing proliferation and differentiation.


Antigenic and enzymatic changes in infected and transformed human diploid cells. Human diploid fibroblasts, WI-38, were infected with various agents and the levels of lysosomal enzymes determined by immunochemical quantitation. Esterase levels were raised by mumps virus and Toxoplasma gondii infection. The concentration of beta-D-glucuronidase was reduced by these same agents. Beta-D-N-acetyl glucosaminidase was greatly increased in cultures infected with T. gondii and decreased by mycoplasma infection. Two cultures transformed by SV40 showed reduced levels of esterase compared with WI-38, as did one of two transformed amnion cultures. A second amnion culture and a culture of transformed Detroit 551 fibroblasts were unchanged. The levels of acid phosphatase were sharply reduced in three of the four SV40 transformed cultures tested.


The interaction of antibodies to two myelin proteins. Antibody has been prepared to a hydrophobic myelin protein in rabbits using several different adjuvants. This antibody was found to react with the myelin basic protein. This latter activity could be absorbed out with basic protein leaving the reactivity to the hydrophobic protein, suggesting that the two myelin proteins shared a common antigenic determinant. In addition, the hydrophobic protein appears to possess a unique antigenic determinant.


Localization of a protein antigen in the chicken spleen. Effect of various manipulative procedures on the morphogenesis of the germinal centre. The time course of the localization of a protein antigen human serum albumin (HSA) into the chicken spleen after intravenous injection is analysed. Localization within seconds to the region surrounding the Schweigger-Seidel sheaths is accomplished by HSA complexes with chicken anti-HSA or by heat aggregated HSA. The localization of soluble HSA has to await the synthesis of sufficient chicken anti-HSA to accomplish localization to the same white pulp sites in the spleen at 25-30 hours after injection. By the use of complexes of HSA-anti-HSA in ten times antigen excess, the time for localization of HSA withing germinal centres was accelerated as compared with soluble HSA, so that newly formed centres containing antigen-bearing dendritic ells were seen at 48 hours instead of 72 hours after use of soluble HSA. Neonatally bursectomized and irradiated (Bx+Irr.) birds fail to localize HSA into germinal centres or to dendritic cells within the white pulp. Heat-aggregated  human gamma-globulin (HGG) injected intravenously into Bx+Irr. birds rapidly localizes within seconds to the periphery of Schweigger-Seidel sheaths and at 24 hours can be seen attached to the surface of typical dendritic cells throughout the white pulp. Hence, heat-aggregated HGG can localize to dendritic cells in the absence of specific antibody. However, such localization to dendritic cells in Bx+ Irr. birds is not followed by segregation of the aggregated HGG-bearing dendritic cells within germinal centres--a further stage in the process which is presumed to require B cells and/or specific antibody. Localization of heat-aggregated HGG to white pulp dendritic ells was prevented by treatment with pepsin sufficient to destroy the ability of aggregated HGG to activate guinea-pig complement. Similary, in vivo decomplementation with a purified anticomplementary fraction (CoF) from the venom of Naja naja resulted in failure of intravenously injected HSA to localize to white pulp dendritic cells and failure of subsequent germinal centre formation. However, such decomplementation did not prevent the localization of aggregated HGG to white pulp dendritic cells. These facts are discussed in the light of hypotheses concerning germinal centre formation and the homeostasis of the antibody response in the bird.


Effects of complement depletion in experimental chagas disease: immune lysis of virulent blood forms of Trypanosoma cruzi. In mice infected with virulent blood (trypomastigote) forms of Trypanosoma cruzi, complement depletion with cobra venom factor caused a marked exacerbation of the disease evidenced by significantly increased parasitemia levels and early mortality as compared with those of untreated infected animals. The effect was greater in mice receiving cobra venom factor on day 7 postinfection, i.e., at the time when the parasites had had time to localize and multiply in the tissues and appeared in the circulation in appreciable numbers. The possibility that complement participates in host defense against T. cruzi infection through a mechanism involving immune lysis was explored in vitro. T. cruzi trypomastigotes were found to undergo immune lysis in sera of patients with chronic Chagas' disease, in sera of immunized mice, and in solutions containing both immune mouse gamma globulin and a source of active complement. This phenomenon failed to take place either in the absence of complement or after complement inactivation by heat or utilizing complement inactivators. The lytic capacity of heated sera was restored by the addition of active complement to the system. During the immune lysis of T. cruzi blood forms, complement was activated in human sera via both the classical and the alternate pathways. In mouse sera, activation followed at least the alternate pathway.


Quantification of mouse macrophage chemotaxis in vitro: role of C5 for the production of chemotactic activity. Delineation of the mechanisms of macrophage accumulation at local tissue sites will further our understanding of immunologically mediated host resistance to infectious and neoplastic diseases. Since mice are frequently used for the study of immune function, we developed a method for the quantification of mouse macrophage chemotaxis in vitro. By this method it was found that the fifth component of complement is necessary for the production of chemotactic activity in mouse serum by inflammatory agents such as endotoxin or aggregated gamma globulin. The majority of macrophage chemotactic activity produced by these agents in mouse serum can be attributed to a low-molecular-weight (ca. 15,000) chemotactic factor. The data suggest that this factor is the biologically active cleavage product of the fifth component of complement, C5a.


Attachment and ingestion of gonococci human neutrophils. Previous studies have indirectly shown that type 1 gonococci are more resistant to phagocytosis by human neutrophils (PMN) than type 3 gonococci. Using phase contrast, fluorescent, and light microscopy, we directly quantitated PMN-gonococcal interaction, with emphasis on separating ingestion from attachment. PMN monolayers were incubated on slides with type 1 or type 3 gonococcal fluorescent antibody (FA). After methanol fixation, the FA-stained gonococci associated with PMN were cointed. Since the live PMN excludes FA, the FA-stained gonococci represent only extracellular gonococci. Methylene blue was then added to the smae slide to stain both ingested and surface attached gonococci. Using these methods, intracellular and extracellular cell-associated gonococci were quantitated under varying conditions. The numbers of methylene blue-stained cell-associated gonococci that were ingested were: with normal serum, 3.7 plus or minus 4.1 per cent for type 1 and 56.2 plus or minus 3.7 percent for type 3 (P smaller than 0.001); with heat-inactivated serum, 1.0 plus or minus 3.0 per cent for type 1 and 52.6 plus or minus 3.7 per cent for type 3 (P smaller than 0.001); with higher-titer anti-gonococcal antibody serum, 4.8 plus or minus 4.3 percent for type 1 and 64.0 plus or minus 1.6 per cent for type 3 (P smaller than 0.001). Thus, most type 3 organisms were ingested, but most type 1 gonococci were bound on the PMN surface.


Superficial antigens of Campylobacter (Vibrio) fetus: characterization of antiphagocytic component. A glycoprotein responsible for the antiphagocytic properties of Campylobacter fetus has been identified by comparing cells of a wild-type strain with those of a mutant lacking this substance. The antiphagocytic component is demonstrable through electron microscopy as a discrete, negatively charged structure on the periphery of the cell. CLT is readily removed from the cell by mild extraction procedures and contributes to the inagglutinability in O antiserum normally displayed by C. fetus. Cells possessing this antigen are refractory to ingestion by macrophages except in the presence of specific antiserum. In its absence maximum phagocytosis occurs without a requirement for opsonins. It is concluded that the antiphagocytic component comprises a critical virulence factor of the bacterium.


Stimulation of phagocytic activity in the reticuloendothelial systems of mice by lipid A complexed with homologous or heterologous proteins. Heterologous proteins are more suitable carriers than homologous proteins for the expression of endotoxic activity of lipid A prepared from the lipopolysaccharides of Salmonella typhimurium SL1102 (Re form) to stimulate the reticuloendothelial system of mice.


Histamine release from human leukocytes when stimulated by Mycoplasma salivarium. Incubated mixtures of Mycoplasma salivarium and nonsensitized mixed leukocytes released histamine. Spectrophotofluorometric detection of histamine occurred at physiological temperature and pH independent of complement and specific antiserum.


Half-lives of two types of rat homocytotropic antibodies in circulation and in the skin. The half-lives of two classes of rat immunoglobulins with homocytotropic properties, i.e., IgE and IgG2a, in the circulating blood and in the skin were studied. The catabolism of both normal (reaginic) and pathological (myeloma) IgE proteins in circulation was found to be extremely rapid with a half-life of 12 h. In contrast, the half-life of IgE antibody in the homologous skin was calculated to be 7.4 days. On the other hand, the half-life of IgG2a in circulation was about 5 days regardless of normal or pathological origin. IgG2a protein was, however, rapidly cleared from the injected skin site with a half life of 2.4 days, a value not longer than that obtained with nonskin-sensitizing goat IgG. The results indicate that rat IgE has an extremely short half-life in circulation despite of its sensitization period in tissues, and that the affinity for the target cells, as well as the mode of sensitization, of the two classes of homocytotropic antibodies, IgE and IgG2a, is different.


Inhibition by derivatives of phloretin of anaphylactic histamine release from human lung tissue and of prostaglandin F2alpha-induced bronchoconstriction. Derivatives of phloretin (25-1,000 mug/ml) among them polyphloretin phosphate (PPP), inhibited in a dose-dependent manner anaphylactic (birch pollen or horse dander) histamine release from human lung tissue passively sensitized with reaginic serum. Pretreatment with PPP of lung tissue sensitized both to birch pollen and horse dander counteracted to a similar extent the release of histamine induced by either allergen administered in sequence. The phloretin derivatives also antagonized the constrictor action of prostaglandin F2alpha on isolated human bronchi at concentrations which did not impair the responses to histamine. The low and high molecular weight derivatives of phloretin were comparably active on a weight basis in both experimental systems.


Comparsion of the properties of two antigen-induced guinea pig lymphokines. The properties of macrophage migration inhibition factor (MIF) and mitogenic factor (MF) were compared using culture supernatants of antigen-stimulated lymph node cells from inbred guinea pigs. Gel filtration on Sephadex G-100 indicated molecular weights of about 60,000 and 25,000 for MIF and MF, respectively. The lymphokines also differed with respect to heat sensitivity, MIF being largely inactivated by 60 degrees C for 20 min, whereas MF was unaffected by this treatment. The time course of production, antigen specificity of induction, and susceptibility to neuraminidase were also examined.


Chemistry of allergens. XXII. Isolation and characterization of three new antigens from the dialysates of six successive pepsin hydrolyses of beta-lactoglobulin. Twelve new antigens previously have been demonstrated in the pepsin digests of milk proteins. The term "new antigen" is defined as an antigen with a specificity distinct from that of the protein from which it was generated. Three new antigens (alpha-, beta-, and D2i) polypeptides and one nonantigenic gamma-polypeptide have been isolated from the dialysates of six successive pepsin hydrolyses of beta-lactoglobulin. The alpha- and beta-polypeptides were 1/5 and D2i was 1/10-1/20 as potent immunogens as precursor beta-lactoglobulin as determined by the Schultz-Dale technique. The minimum observed amounts of new antigen eliciting maximum response of uterine strips in the Schultz-Dale tests were: alpa-D2, 50 ng; beta-D2, 15 ng; beta-D3; 10 ng; and D2i, 1,000 ng. Except for the alpha-polypeptide, the amino acid contents of the polypeptides differed markedly from that of beta-lactoglobulin. The beta-polypeptide has been tentatively identified as a 33 amino acid fragment of beta-lactoglobulin (3,910 daltons). The gamma-polypeptide has been tentatively identified as a 12 amino acid fragment of beta-lactoglobulin (1,372 daltons).


Determination of serum C9 level by immunodiffusion. Elevation in patients with infectious or allergic skin diseases. The measurement of the ninth component of complement (C9) in human serum can be easily carried out by the Mancini method using antiserum raised against purified C9. The average C9 level in 29 healthy adults was 44.5 plus or minus 10.6 mug/ml. The serum C9 level was often elevated in patients with infectious or allergic skin diseases. The C9 level remained normal in most patients with collagen diseases. The availability of the monospecific antiserum has permitted identifying C9 in human serum as alpha2-globulin.


Studies in immunodermatology. VI. IF studies of autoantibodies to the stratum corneum and of in vivo fixed IgG in stratum corneum of psoriatic lesions. Indirect immunofluorescent (IF) tests on sections of normal human skin reveal the presence of antibodies to the stratum corneum in most normal human sera. Sera absorbed with hyperkeratotic scales gave negative reactions. These reactions appeared to correspond to those of the stratum corneum antibodies which were first detected by Krough and Tonder by immune adherence, though this remains to be documented since the conjugate alone stained the stratum granulosum in a pattern comparable to that of the IF staining reported by Krough. The staining was caused by conjugates with high but not by those with low fluorescein to protein ratios. In indirect IF tests, comparable titers of stratum corneum antibodies were found in sera of psoriasis patients and control subjects. Since they reacted with the stratum corneum of the antibody producer they are referred to as autoantibodies. Direct IF tests of psoriatic lesions revealed the presence of in vivo bound IgG as well as other immunoglobulins and complement in the stratum corneum. To differentiate staining with conjugated protein from direct and indirect IF staining of the stratum corneum a four-compartment test was devised. Skin sections were treated with saline or with stratum corneum antibodies and then with conjugates without (compartments 1 or 2) or with (compartments 3 or 4) anti-IgG antibodies. Sections were read by the method of Kawamura and his associates with both UV and BV illumination. Compartment 3 is a direct IF test and compartment 4 is an indirect IF test. In four-compartment tests performed on both biopsies of psoriatic lesions or scales the difference between compartments 1 or 2 and 3 affords a measure of in vivo bound IgG and the difference between compartments 3 and 4 provides information on the presence of free stratum corneum antigen. With this four-compartment test system it was found that in vivo fixation of IgG occurs in the intercellular areas of the stratum corneum of psoriatic lesions or scales and that this corresponds to the site of binding of the stratum corneum autoantibodies. This in vivo binding of IgG in the stratum corneum of psoriatic lesions resembles somewhat the intercellular fixation of IgG in the stratum spinosum in pemphigus. In the materials examined in these preliminary studies most, if not all of the stratum corneum antigen appeared to be covered with in vivo bound IgG.


Studies in immunodermatology. VII. Four-compartment system studies of IgG in stratum corneum and of stratum corneum antigen in biopsies of psoriasis and control dermatoses. Biopsies of skin lesions from 50 cases of psoriasis and from 42 cases with other dermatoses were studied by the methods described in the foregoing report, that is by the four-compartment by the methods described in the foregoing report, that is by the four-compartment immunofluorescent (IF) system for detecting stratum corneum autoantibodies and in vivo binding of IgG in psoriatic scales at the site of the stratum corneum antigen. They were also studied histologically. Early and fully developed psoriatic lesions tended to have more widespread deposits of IgG in the stratum corneum than receding lesions. In biopsies with widespread IgG deposits the stratum corneum antigen usually could not be demonstrated because of blocking by in vivo bound IgG. Of the 42 control biopsies of skin lesions from patients with other dermatoses which were examined with this system two gave the IF staining patterns which were seen most commonly in psoriatic lesions. In these two cases strong widespread IgG deposits occurred in compartment 3, and 4 was very similar to compartment 3. One case was diagnosed as lichen planus and one as cornu cutaneum. Comparisons of the IF findings in psoriatic and control lesion biopsies indicate that significantly more of the former have: (1) IgG deposits at the combining sites of stratum corneum antibodies (92 vs. 50%) and (2) no demonstrable stratum corneum antigen because of 'blocking' by in vivo bound IgG, i.e. compartment 3 like 4 (56 vs. 7%). A highly significant inverse relation between the in vivo deposits of IgG and demonstrability of stratum corneum antigen appeared regardless of the clinical condition. This observation lends support to the view that the IgG which is bound to the sites of the stratum corneum antigen in ski; lesions may in fact be in vivo bound stratum corneum antoantibody.


Lymphoid cells in infectious mononucleosis classified according to T and B cell markers. It has been demonstrated that peripheral blood lymphocytes, particularly the "atypical" ones, are predominantly of the T type in infectious mononucleosis (IM). This is based on membrane marker studies (E rosettes, receptor for complement, receptor for Fc fragment of immunoglobulins (Ig), and membrane Ig) and by anti-T lymphocyte serum. On the other hand, lymphoblastoid cell lines derived from IM patients show the characteristics of B lymphocytes. This permits the suppostion that EBV infects B lymphocytes and stimulates them to proliferate. The cell proliferation is unlimited in vitro and is probably controlled in vivo by T cells. The reduction of cellular immunity in vivo, which contrasts with the high number of T cells in peripheral blood, could be explained by the fact that T cells are engaged in the regulation of B cell proliferation.


Observations on rabbit thymocytes and peripheral T cells. I. Anomalous results in the mixed antiglobulin reaction caused by non-specific adsorption of sheep immunoglobulin. The "single-stage" mixed antiglobulin reaction (MAR) was carried out with rabbit thymocytes. This test involved treating the cells with either sheep or goat anti-rabbit globulin sera, and subsequently reacting them with indicator erythrocytes coated with rabbit immunoglobulin (Ig) so as to form rosettes. An unexpectedly high number (up to 38%) of thymocytes reacted, although the rosettes were weaker than those given by peripheral B lymphocytes. When blood and lymph node lymphocytes or thymus cells which had already been treated with sheep anti-rabbit globulin serum were subsequently exposed to rabbit anti-sheep Ig serum and then rosetted with indicator cells coated with ox Ig (cross-reacts with sheep Ig) almost 100% reaction was obtained in each of the cell suspensions. This was designated the "two-stage" MAR. The anomalous results, both in the one-stage and two-stage MAR, were abolished by pepsin-treating the sheep anti-rabbit globulin serum; thus indicating that sheep Ig is adsorbed non-specifically via the Fc part of the molecules to the surface of rabbit thymocytes and peripheral T lymphocytes.


Immune response in the guinea pig to penicillin-autologous carrier proteins. The spontaneous fixation of penicillin G on guinea pig serum protein was studied. Serum albumin and Ig were shown to fix firmly over 95% of penicillin. Therefore, guinea pigs were immunized with penicillin alone or penicillin-autologous carrier protein conjugates. The production of hemagglutinating antibody, of MIF and the in vitro stimulation of lymphocytes were analyzed as immunological parameters. The results showed that in all cases the strongest immunological response was obtained with animals immunized with the protein-hapten conjugates, whereas no response could be obtained with the carrier protein alone. Finally it was shown that the conjugates significantly stimulated purified T cell population. These results suggest that the antigenic recognition of penicillin involves the participation of lymphocytes.


Ultrastructural localization of L-alpha-hydroxy acid oxidase in rat liver perioxisomes. The localization of L-alpha-hydroxy acid oxidase in rat liver peroxisomes was studied using slight modifications of the Shnitka and Talibi (1971) method. Best results were obtained with formaldehyde fixation and incubation with glycolate as substrate. Following incubation the copper ferrocyanide reaction product was amplified with 3,3'-diamino-benzidine according to Hanker et al. (1972a,b). Dense reaction product was visible in hepatocyte peroxisomes by light and electron microscopy. Some diffusion of enzyme and/or reaction product into the adjacent cytoplasm occurred around the peroxisomes. Apparent non-specific deposits occurred on the plasmalemma, in the nucleus, and occasionally over mitochondria. Glutaraldehyde fixation severely inhibited enzymatic activity, and the enzyme showed less activity toward L-lactate and DL-alpha-hydroxybutyrate.


The demonstration of arylsulfatases with 4-nitro-1,2-benzenediol mono(hydrogen sulfate) by the formation of osmium blacks at the sites of copper capture. A new method is described for the direct cytochemical demonstration of lysosomal arylsulfatases utilizing a synthetic substrate, 4-nitro-1,2-benzenediol mono(hydrogen sulfate), and a copper capture reaction. A small amount of Hatchett's brown (cupric ferrocyanide, Cu2Fe(CN)6-7 H2O) formed at the subcellular sites of copper capture is then utilized as a heterogeneous catalyst to effect the oxidative polymerization of 3,3'-diaminobenzidine which results in the formation of an insoluble, highly colored osmiophilic indamine polymer at the sites of enzymatic activity. The reaction product even at this stage prior to osmication is highly visible. It is readily seen with a light microscope in 50 mum sections of fixed tissues prepared with a mechanical chopper or in 10 micron cryostat sections treated for arylsulfatase activity. Upon osmication, an electron-opaque osmium black is formed which is much less soluble than the products of either the lead or barium capture reactions currently used for the demonstration of arylsulfatase with the electron microscope. The selection of areas of plastic-embedded tissues for ultrathin sectioning is facilitated by the ready visibility of these osmium black end products on 1-2 mum plastic sections which can be studied with the light microscope. This method gives permanent specimens demonstrating arylsulfatases A or B in lysosomes and autophagic vacuoles. In addition, enzyme activity is seen occasionally in the Golgi region or lamellae of certain cells believed to be elaborating sulfated products. In these instances, it may be demonstrating sulfotransferase activity.


Use of the analytical electron microscope (AEM) in cytochemical studies of the central nervous system. Central nervous tissues (median eminence and arcuate nucleus) were studied by means of energy dispersive x-ray analysis using electron optical systems (analytical electron microscopy). These studies were conducted after the tissue had been treated specifically for localized biogenic amines (BA). The results indicate that not only is the BA cytochemical reaction highly specific, and that BAs can be localized intraneuronally in areas not previously identified, but also that the analytical electron microscope is a very valuable and potentially powerful tool in the studies of inclusion bodies and organelles in the central nervous system and other tissues. Thus, the nonspecific density production by osmium tetroxide can be elucidated from the specific BA reaction plus other areas of the nervous system containing density producing heavy metals, i.e., iron are readily identifiable.


Optical versus radiographic magnification for fine-detail skeletal radiography. Fine-detail radiographic techniques for peripheral skeletal imaging have gained wide clinical acceptance. In this study, the imaging properties and clinical applications of the optical magnification technique, which employs fine-grain industrial film and a large focal spot, are compared quantitatively and qualitatively with those of three slow screen-film techniques, namely, contact exposure with a large focal spot, 2 times radiographic magnification with a 0.3 mm focal spot, and 4 times radiographic magnification with a 50 mu focal spot. The modulation transfer functions (MTF's) of the recording systems and focal spots are obtained and film sensitometry performed. Clinical comparisons are made for patients with metabolic, arthritic, and neoplastic skeletal disorders. The results illustrate the superiority of the optical magnification technique over contact or 2 times magnification techniques using slow screen-film systems. If a microfocus tube is used, however, direct radiographic magnification may provide images comparable in resolution, noise and contrast to those made with the optical magnification technique, and at lower radiation exposure to the patient.


Observations on the selective demonstration of nucleolar material by protein staining techniques in epon thick sections. The effect of different protein stainings on thick sections of Epon embedded salivary glands of Chironomus pallidivittatus is described. Pretreatment of sections with several metallic salts affected the affinity for aniline blue black in different structures. The uranyl and aluminium ions followed by aniline blue black resulted in a highly selective nucleolar staining which proved also efficient in Allium cepa meristematic cells and which seems useful for the preferential demonstration of nucleolar material in thick sections.


The influence of fixation on staining of glycosaminoglycans in glial cells. Cytoplasmic staining of glial cells by Alcian blue in 0.40 M magnesium chloride can be demonstrated in sections of brain tissue fixed in formol-acetic acid and in Bouin's solution. Phosphate-buffered aldehyde fixatives, whether at neutral or low pH, fail to preserve stainable material.


Fine structure of Methanospirillum hungatii. The fine structure of Methanospirillum hungatii was studied by electron microscopy. The topography of the cell wall and the mechanism of cell division are not typical of gram-positive or gram-negative bacteria. A novel architectural arrangement of cells in continuous spiral filaments is described. Filamentous cells are connected by spacers and enclosed within a rigid outer envelope. The unique ultrastructural features of cells and cell spacers were examined.


Precursor relationship of phenylalanine transfer ribonucleic acid from Escherichia coli treated with chloramphenicol or starved for iron, methionine, or cysteine. When treated with chloramphenicol, Escherichia coli 15T minus produces two new species (IV and V) of transfer ribonucleic acid specific for phenylalanine in addition to the major normal species (II) and two minor normal species (I and III), which are seen as distinct components upon fractionation by chromatography on columns of benzoylated diethylaminoethyl-cellulose. Species IV is produced when cells are grown in iron-deficient medium and is, therefore, probably deficient in the 2-methylthio modification of N-6-(delta-2-isopentenyl) adenosine. A new minor species (Va) also appears under those conditions. All of the new components elute earlier than the major normal species. Addition of chloramphenicol to iron-deficient cells leads to the production of species V, and that production is blocked by rifampin, as is the production of species IV. Thus, species IV and V appear to be transcriptional products. Although E. coli 15T minus appears to be rel plus, starvation for methionine or cysteine leads to the accumulation of species IV (without addition of chloramphenicol); rifampin blocks the accumulation. Species V is still produced on addition of chloramphenicol to starved cultures. Starvation for arginine or tryptophan does not alter the chromatographic profile from the normal case. Treatment with permanganate indicates that species II and IV contain isopentenyladenosine but that species V does not. Species V appears to be deficient in both isopentenyl and methylthio modifications of adenosine and perhaps at least one other modification, because removing the isopentenyl moiety from adenosine does not convert species IV into species V, but converts it into species Va. A precursor relationship among species V, VI, and II is suggested by following the chromatographic profile of phenylalanine transfer ribonucleic acid during recovery of E. coli from treatment with chloramphenicol; the various species increase and decrease in a sequential manner.


Regulation of acetohydroxy acid synthase in streptomycin-dependent Escherichia coli. Growth of streptomycin-dependent mutants of Escherichia coli K-12 was insensitive to valine when dihydrostreptomycin was present in a nonlimiting concentration in glucose-salts medium. Acetohydroxy acid synthase was derepressed under these conditions, owing to relaxation of catabolite repression. Valine sensitivity and catabolite repression were restored when streptomycin-dependent E. coli K-12 mutants were grown with limiting dihydrostreptomycin. End product repression of acetohydroxy acid synthase under conditions of relaxed catabolite repression was effected by any two (or more) end products except the combination valine plus isoleucine, which caused derepression. Single end products had no detectable effect on acetohydroxy acid synthase formation.


Results of treatment of spinal tuberculosis by "middle-path" regime. The efficacy of modern drugs in the treatment of tuberculosis of the spine has been evaluated by a personal follow-up for three to ten years. Operation on the vertebral lesion was done only for those patients with or without neural complications who failed to respond favourably to drug therapy and rest. Thus absolute indications for operation were present in only 6 per cent of cases without neural involvement and in 60 per cent of patients with neural deficit. Of patients who responded to drug therapy alone, only 19 per cent revealed increase of kyphosis by more than 10 degrees. The diseased area showed radiological evidence of osseous replacement of 29-6 per cent of cases, of fibro-osseous union in 50 per cent and of fibrous replacement in 20-2 per cent. The overall results of this regime compare favourably with those of radical operation. It is suggested that treatment should in the first place be by modern antitubercular drugs.


Protein migration into nuclei. I. Frog oocyte nuclei in vivo accumulate microinjected histones, allow entry to small proteins, and exclude large proteins. A technique is presented which enables one to measure the extent to which a protein enters and accumulates in the nucleus of the frog oocyte. In this method, the protein, labeled with 125-I, is microinjected into the oocyte. After incubation, the oocyte is manually enucleated and the radioactivity in the nucleus and cytoplasm is determined. Using this technique, proteins lighter than 20,000 daltons were found to enter the nucleus and completely equilibrate between the nucleus and cytoplasm within 24 h. The entry of proteins heavier than 69,000 daltons was severely hindered. Histones and histone fractions entered as quickly as other small proteins, but, in contrast to these proteins, they accumulated in the nucleus to different extents, depending on the total amount of histone injected into the oocyte and the identity of the histone. Evidence is presented that histone fractions compete with each other for accumulation in the nucleus.


Development and validation of a radioimmunoassay for peptides related to beta-melanocyte-stimulating hormone in human plasma: the lipotropins. A radioimmunoassay is described for the measurement of human "beta-melanocyte-stimulating hormone" ("betah-MSH"). Two antisera have been used, one of which cross-reacts with synthetic betah-MSH as well as with the two larger pituitary peptides betah- and gammah-lipotropin (betah- and gammah-LPH) and the other mainly with betah-MSH and gammah-LPH. The sensitivity and reliability of the assay have been improved by employing a simple plasma extraction procedure, and the shelf-life of the iodinated betah-MSH tracer has been increased more than five-fold by storage in a concentrated human serum albumin solution. Using a 5 ml plasma sample the detection limit is 6 pg/ml. The mean resting "betah-MSH" level in normal subjects is 21 pg/ml (range 13-38 pg/ml) at 9 AM and 12 pg/ml (range 6-20 pg/ml) at 9 PM. Levels are considerably elevated (51-12,000 pg/ml) in patients with Addison's disease. Nelson's syndrome, Cushing's disease and the "ectopic" ACTH syndrome. After administration of insulin or pyrogen, the concentration of plasma "betah-MSH" increases in parallel with that of ACTH and they are approximately equivalent on a molar basis. The stability of purified betah- and gammah-LPH and endogenous "betah-MSH" when incubated in vitro in fresh blood or plasma are similar, in contrast to the less stable peptide synthetic betah-MSH. It is suggested that "betah-MSH" immunoreactivity in human plasma is due to betah- and gammah-LPH rather than betah-MSH.


An evaluation of programmed text, slide-tape, and lecture at six dental schools. The results of previous studies reported in the literature have been inconclusive as regards the effectiveness of programmed instruction as compared to the lecture method. The results of this study demonstrate that the formats are, in the end, of similar effectiveness. Therefore, the choice of teaching format in a dental school can safely be based on considerations other than the amount of learning that will occur.


Effects of aging on granulopoietic activity (colony-stimulating factor). This investigation was undertaken to determine whether there is a decrease of granulopoietic activity in subjects of the geriatric age group. Colony-stimulating factor (CSF) is an alpha glycoprotein which causes proliferation of granulocytes in vitro. A study was made of the variation in the level of CSF with age and the state of health in 78 subjects. They were classified into two groups-young (ages 22-60) and old (ages 70-94). The state of health was classified as normal, acute disease, or chronic disease. The results showed that serum CSF does vary with age and health. The CSF levels were higher in the young than in the old. For both age groups, the CSF levels were elevated in acute disease, and subnormal in chronic disease.


Effects of repeated oral doses of demethylchlortetracycline on bones and dentin of young rhesus monkeys. Seven oral doses of demethylchlortetracycline were administered to monkeys that received serial injections of lead acetate to intravitally stain calcification sites. Bone growth was greatly inhibited, whereas dentin apposition was spared from the cumulative toxicity of demethylchlortetracycline. Cessation of bone growth and its duration could be correlated with serum levels.


Comparison of Sirius red and Congo red as stains for amyloid in animal tissues. Sirius red and Congo red were compared for specificity and sensitivity of amyloid staining in animal and human material. Previously described advantages of Sirius red as an amyloid dye were confirmed, as well as its disadvantage of lack of ultraviolet fluorescence. Two further disadvantages of Sirius red were discovered, both relating to animal material: (a) its unexpectedly weak staining of early experimentally induced amyloid deposits and (b) frequent uncontrollable nonspecific staining of fibrous tissues. It is therefore concluded that, overall, Congo red used by the improved alkaline technique of Puchtler, Sweat and Levine (1962) remains the best single method for demonstration of amyloid in both human and animal tissues.


Abnormal mast cell granules in the beige (Chédiak-Higashi syndrome) mouse. Mast cells of beige (C57BL/6J) (bg-j/bg-j) mice were examined histochemically and ultrastructurally. Mast cell granules in the beige mice were markedly enlarged and irregular in shape. Granule contents stained uniformly with acidified toluidine blue, but with ruthenium red and Alcian Blue-safranin, two components were evident. The rims of the abnormal granules stained with ruthenium red and with Alcian Blue; the centers of the granules were clear with ruthenium red and stained with safranin. Mast cell granules thus represent another abnormal organelle in the Chédiak-Higashi syndrome.


Adoptive transfer of experimental allergic encephalomyelitis (EAE): prevention of successful transfer by treatment of donors with myelin basic protein. After onset of experimental allergic encephalomyelitis (EAE), guinea pigs can be effectively treated by injection with myelin basic protein (BP). In order to localize the site of action of BP, cells from sensitized donors treated with BP one, two, three, or four times after disease onset have been transferred to normal recipients. One injection of BP has no effect on ability of cells to transfer EAE. Two injections partially inhibit transfer. After the third and fourth injections the sensitized cells lose their capacity to transfer EAE. The therapeutic effect of BP previously demonstrated in actively sensitized guinea pigs must involve the specifically sensitized cells rather than the target organ.


Immunofluorescent studies on antibodies directed to a buried membrane structure present in lymphocytes and erythrocytes. Brief digestion of human peripheral blood lymphocytes by vibrio cholera neuraminidase (VCN) revealed hidden components of the membrane. Autologous human serums contained antibodies directed to these components that were readily demonstrated by immunofluorescence. Antibodies of similar specificity were found in all normal serums. The antibodies were principally of the IgM variety with lesser amounts of the IgG class present. They were equally active at 4 degrees C and 37 degrees C. The VCN revealed membrane determinants were present in normal B and T lymphocytes, monocytes, lymphocytes of patients with chronic lymphatic leukemia and cells of lymphoid lines. The newly revealed determinants slowly disappeared upon culture of the lymphocytes. These hidden components were similarly demonstrated in erythrocyte membranes and represent the T antigen long known for the red blood cells. Absorption by either VCN treated autologous lymphocytes or erythroyctes removed all of the antibodies capable of reacting with both cell types. Absorption by VCN digested isologous lymphocytes removed all reactivity with autologous lymphocytes.


Immune specific production of interferon by human T cells in combined macrophage-lymphocyte cultures in response to Herpes simplex antigen. Human peripheral blood lymphocytes, highly enriched for T cells, were obtained by passing gravity-sedimented leukocytes through nylon wool columns. The eluted cells were cultured with autologous macrophages and the mixture was studied for its capacity to produce interferon in vitro in response to stimulation with herpes simplex virus antigen. The interferon produced by the combined macrophage-lymphocyte cultures was shown to depend upon the presence of T cells; elimination of these cells by treatment with an anti-T cell serum plus complement greatly diminished the amount of interferon produced. The memory for the immune-specific release of interferon also appeared to be carried by the T lymphocytes rather than the glass-adherent macrophages. Furthermore, the results suggest that under our conditions of culture immune-specific interferon originates from T cells.


Characterization of human B lymphocyte specific antigens. Anti-human B cell serum (ABS) was developed by sequentially absorbing a rabbit anti-human tonsil serum (AHTS) with human red cells, liver, serum, and thymocytes. AHTS was also absorbed quantitatively with tissues and cells from different sources. ABS was nontoxic for thymocytes but lysed the majority of chronic lymphocytic leukemia (CLL) cells. This also killed a number of tonsil and blood lymphocytes which bound erythrocyte-antibody-complement complexes but not sheep erythrocytes. In further studies, it was shown that phytohemagglutin-responsibility of peripheral blood lymphocytes was not affected by treating them with ABS and complement. Anti-B cell activity of AHTS was not changed by absorption with thymus, liver, kidney, and brain tissues but absorbed with tonsil and CLL cells. These data confirm the B cell specificity of ABS. Indirect immunofluorescence was carried out on tissue sections of human tonsils and lymph nodes, which indicated that ABS-reactive cells were concentrated in lymphoid follicles including germinal centers, while plasma cells and cells located in the thymus-dependent area were essentially devoid of immunofluorescence.


Strain differences in ease of tolerance induction to bovine gamma-globulin: dependence on macrophage function. DBA/2 mice are highly susceptible while BALB/c mice are resistant to the induction of tolerance with ultracentrifuged BGG. As measured by the clearance of a test dose of 125-I-BGG, 2 mg ultracentrifuged BGG is sufficient to induce complete tolerance in DBA/2 mice, while BALB/c mice exhibit a clearance rate similar to immune controls. BALB/c mice pretreated with carrageenan (a macrophage toxic agent) lose their resistance to tolerance induction with 2 mg ultracentrifuged BGG, while treatment with cobra venom factor does not alter their resistance to tolerance induction. Stimulation of the reticuloendothelial system with BCG produces a reduction in the susceptibility to tolerance of DBA/2 mice. This effect is transitory, with the maximum effect 3 weeks after BCG infection. Biologic filtration of BGG through BALB/c mice results in a preparation which is highly tolerogenic for recipient mice and this ability to filter the immunogenic portion of BGG is radioresistant. The results provide evidence for the concept that the differential susceptibility to the induction of tolerance is related to a macrophage function in the inductive phase of immunity.