[Evidence and meaning of monophasic potentials in the electromyogram of extraocular muscles of rabbits and cats (author's transl)]. The EMG of extraocular muscles of rabbits and cats was recorded by a concentric-needle electrode provided for clinical use. The EMG showed spike activity (at least biphasic, fast waves of 1 to 2 msec duration) and characteristic long-duration monophasic waves (decline for up to 7 msec). The monophasic potentials are interpreted as local depolarizations arising at the nerve-muscle junctions of multiply innervated muscle fibers. Spike- and monophasic potentials are discharged in independent rhythms whose frequency changes when the head position is altered. The frequency of spike- and monophasic potentials are changed in the same direction by labyrinthine reflexes. Spontaneous monophasic potentials recruit at a lower threshold.


In vitro estrone-estradiol-17beta interconversion in the cornea, lens, iris and retina of the rabbit eye. Estrone and estradiol-17beta were incubated with various eye tissues of female rabbits. The interconversion of both estrogenes by the action of 17beta-hydroxysteroid dehydrogenase decreased in the sequence cornea, iris, retina, and lens. In a smaller yield estradiol-17alpha was also formed from estrone. The cornea seems to be an active estrogen-metabolizing tissue. The role of estrogens and progestogens in the tissues of the anterior eye segment is briefly discussed.


Physical properties of sputum. VII. Rheologic properties and mucociliary transport. Sputums from bronchitic patients were measured for viscosity and elastic recoil and then placed on an isolated frog pharyngeal palate to study the velocity of transport by beating mucosal cilia. Purulent sputum, as a group, demonstrated a higher viscosity and a lower elastic recoil, and was transported at a slower rate than mucoid samples of low viscosity and high recoil. Indigenous frog mucus, collected and reapplied to the palate, had the lowest viscosity, the highest elastic recoil, and the fastest transport rate. Thus, it was concluded that high elastic recoil and low viscosity represent the best rheologic combination for maximal velocity by a mucociliary system.


Rapid detection of ampicillin-resistant Haemophilus influenzae and their susceptibility to sixteen antibiotics. Ampicillin-resistant and -susceptible strains of Haemophilus influenzae were tested for susceptibility to 16 antibiotics. Chloramphenicol and a new cephalosporin, cefamandole, were most active with minimal inhibitory concentrations (MICs) for all bacteria tested between 0.5 to 2.0 mug/ml. All but two organisms were susceptible to tetracycline. Ampicillin-resistant strains of H. influenzae were less susceptible (MIC, 4 to 32 mug/ml) to carbenicillin and ticarcillin than ampicillin-susceptible organisms (MIC, 0.25 to 1.0 mug/ml). A rapid assay for beta-lactamase, utilizing a chromogenic cephalosporin substrate, detected enzyme production in all 17 ampicillin-resistant strains of H. influenzae.


Modification of postirradiation hydroproteolytic activity of rat's pancreas by some drugs. Influence of Trasylol and morphine on hydroproteolytic activity of rat's pancreas was investigated. Trasylol, administered in vivo, caused normalization of activity of some pancreatic enzymes in the rats single and fractionated irradiated. In vitro, however, Trasylol did not affect the catheptic (pH 5.4) and amylolytic activity of the pancreas. After in vivo administration of morphine, the return to control values of amylolytic activity in the serum of single irradiated animals was observed.


A rehabilitation evaluation system which complements the problem-oriented medical record. The traditional medical history and physical examination format is disease rather than disability oriented. It has been shown to be incomplete for the total evaluation of rehabilitation patients. Direct applications of Weed's Problem-Oriented Medical Record have proven to be formidable and cumbersome due to the complexity and diversity of rehabilitation. Therefore, we have developed the Rehabilitation Evaluation System (RES) to document functional rehabilitation management and progress during inpatient hospitalization and outpatient follow-up. The system identifies 18 key rehabilitation areas, each with an individual and objective four-point scale. Utilization of this system in our department has been invaluable in formulating goals and continually evaluating the on-going rehabilitation process. We used the RES with equal facility on 46 rehabilitation inpatients including stroke, amputation, spinal cord injury, multiple sclerosis, orthopedic-trauma, rheumatoid arthritis and poliomyelitis. The mechanics of the RES are presented in detail with a specific patient-example of hemiplegia. Its complimentary use with the Problem-Oriented Medical Record is discussed. Practical advantages are seen in patient care, medical student and resident education, record keeping and research.


Unsuspected intravestibular schwannoma.

Unsuspected intravestibular schwannoma. The intravestibular schwannoma is physically minute and will often elude preoperative diagnosis. From time to time, such a tumor will be unexpectedly encountered during transcanal labyrinthectomy. A review of pathologic and surgical literature suggests that the lesion can be well managed by the transcanal route. A standard labyrinthectomy may be sufficient, but the translabyrinthine approach may be necessary for more extensive tumors.


A case of lymphoma with T-cell characteristics. This case report concerns a woman of 54 who died from a rapidly progressive lymphosarcoma-cell leukaemia with the terminal complication of toxoplasmosis.  The grossly atypical lymphosarcoma cells were shown to form rosettes with sheep erythrocytes but were incapable of responding to stimulation with phytohaemagglutinin.  They thus had the functional characteristics of abnormal T-cells.  The proportion of immunoglobulin-bearing lymphocytes was greatly reduced.


Some effects of alcohol on various aspects of oculomotor control. Recent studies have shown that alcohol interferes with visual control of vestibular nystagmus. The present study was designed to assess three partially independent systems of oculomotor control. Performance on three tasks was measured before and after mild alcohol dosage. One task involved visual suppression of vestibular nystagmus; a second involved smooth oculomotor tracking of a moving target; and a third required repetitive rapid voluntary shifts in gaze. Oculomotor control was degraded on the first two tasks with recovery toward the initial performance level 4 h after drinking. Performance on the third task was not obviously degraded, although it is possible that improvement with practice was retarded. Results are discussed in terms of neurological systems involved and kinds of flight tasks potentially affected.


Visualization of surface topology of human lymphoid cells by scanning electron microscopy. We have studied the surface characteristics of human lymphoid cells with the scanning electron microscope (SEM). We obtained thymic cells by mincing thymus and isolating peripheral blood lymphocytes from normal donors as well as patients with acute and chronic lymphocytic leukemia (ALL and CLL). Prior to SEM, we purified the lymphocytes in isoosmotic Ficoll-Hypaque gradients. In addition, we examined several lymphoid cell lines in permanent culture derived from normal persons, patients with infectious mononucleosis, as well as from patients with ALL. The surfaces of most thymic cells appear smooth with only a few short microvilli. A small number of thymic cells, on the other hand, are found to exhibit many more microvilli of uniform shape and length. We could further identify lymphocytes from normal donors as T- and B-types according to their rosetting spontaneously with sheep red blood cells (SRBC) resemble thymic cells in their surface topology. However, B-cells rosetting with amboceptor--and complement--treated SRBC show greater number of microvilli. The surface morphology of lymphocytes from leukemic patients appears more or less characteristic of the disease type. In contrast, the surfaces of cultured lymphoid cells show a diversified surface topology, often with large smooth areas, very long and very short microcilli, which may often cluster in groups. These data bear directly upon techniques of plasma membrane fractionation and we suspect that the topology differences correlate with their differentiation patterns.


Studies on the effector cell of anti-tumour immunity in a chemically induced mouse tumour system. Spleen cells from mice immunized against a methylcholanthrene induced rhabdomyosarcoma inhibited tumour cell colony formation in vitro and prevented tumour development in vivo in an adoptive transfer test. Treatment of the immune spleen cells with anti-mouse immunoglobulin serum or passage through a nylon wool column, both of which reduced the percentage of immunoglobulin bearing cells in the population to less than 3-4%, did not alter their anti-tumour effects. In contrast, treatment of the spleen cells with anti-BAomicron serum abolished their anti-tumour effects both in vitro and in vivo. These results indicate that T cells are the mediators of tumour immunity in this chemically induced tumour system.


The enhancing influence of proteolysis on E rosette forming lymphocytes (T cells) in vivo and in vitro. The T lymphocytes populations of 22 young healthy adults, 21 healthy middle aged and older blood donors, 35 non-pregnant women of child bearing age and 14 patients with advanced malignant disease were assessed and compared. It was found that the mean T cell counts in the middle aged and older controls were significantly lower than in the healthy young adults and were further reduced in the patients with malignant disease. The addition of the proteolytic agent brinase (protease 1 obtained from Aspergillus oryzae) to the rosetting test increased the T cell counts signficantly in all groups. This was mot marked in the older age groups and the patients with malignant disease. The proteolytic agent is shown to exert its effect on the lymphocytes in the test. Slow intravenous infusion of either brinase or streptokinase into patients with malignant disease is shown to result in increased T lymphocyte counts pari passu with a restoration of skin allergy. The significance of these findings and possible mode of action of the proteolytic agents in increasing T cell activity are discussed.


Effect of local injection of Corynebacterium parvum on the growth of a murine fibrosarcoma. Local injection (i.e. injection at the site of tumour inoculation) of strains of C. Parvum which have a significant anti-tumour effect when given systemically (i.e. intravenously or, in the case of subcutaneous tumour transplant, intraperitoneally) strongly inhibits the growth of isogeneic transplants of a fibrosarcoma in intact CBA mice but has little or no effect on subcutaneous transplants of the same tumour in T-cell deprived mice. The anti-tumour effect of local injection of C. parvum, unlike that of systemic injection in this particular tumour system, thus appears to be T-cell dependent.


Stimulation of granulocytic colony formation  in agar diffusion chambers implanted in cyclophosphamide pretreated mice. The growth of mouse bone marrow colonies in agar diffusion chambers (ADCs) was evaluated using host mice injected with saline or with cyclophosphamide (CY) before chamber insertion. The mice pretreated with cyclophosphamide proved more effective hosts than control (saline pretreated) mice, indicating that cyclophosphamide causes the elaboration of a stimulating factor acting on colony precursor cells. Assays of the factor for colony stimulating activity against mouse bone marrow cells in agar culture in vitro suggest that potentiation may be due to a slight temporary increase in the level of colony stimulating factor (CSF) in the chamber environment, although a parallel increase was not detected in the serum. Stem cell recovery from the ADCs, measured by spleen colony formation, suggests that the stimulus may act by increasing differentiation at the level of the pluripotential stem cell.


Storage and metabolism of poly(adenylic acid)-mRNA in germinating cotton seeds. When the RNA from both the nucleus and polysomes of germinated cotton seeds were fractionated on poly(U)-cellulose columns it was found that both contained poly(A)-RNA. In ungerminated seeds most of the poly(A)-RNA is located in the nucleus of the cell. 3'-Deoxyadenosine was found to inhibit the synthesis of both nuclear and polysomal poly(A) by 75% in germinated seeds, but had little or no effect on protein synthesis during the first 6 hr of germination. After this time 3'-deoxyadenosine had a pronounced inhibitory effect on protein synthesis presumably due to the inhibition of poly(A) addition to newly synthesized mRNA.


The functional unit of electrical events and phosphorylation in chromatophores from Rhodopseudomonas sphaeroides. 1. From electron micrographs of chromatophores from Rhodopseudomonas spaeroides and from the estimated bacteriochlorophyll per chromatophore was estimated. The mean diameter of the chromatophore vesicles was 600 A. 2. The decay of the flash-induced electric potential across the chromatophore membrane measured by the carotenoid band shift was 20% accelerated by about one valinomycin molecule per 4700 bacteriochlorophyll, i.e. by one ionophore molecule per chromatophore. 3. The inhibition of the flash-induced ATP formation by valinomycin followed a similar pattern to the accelerated decay of the electric potential. 4. The single turnover flash yield of ATP synthesis gave a mean value of one ATP per 1470 bacteriochlorophyll or about 3 ATP per vesicle. 5. With regard to the partitioning of the ionophore between the membrane (85%) and aqueous phase (15%) we conclude that one molecule of valinomycin per chromatophore is sufficient to begin to collapse the electrical potential and inhibit ATP synthesis. It is therefore suggested that the membrane potential is an essential component of the energized state which is used for phosphorylation. The results correspond to those obtained for the 100-fold larger vesicles in chloroplasts (thylakoids) where one molecule of ioophore is also sufficient to quench both events.


Treatment of life-threatening hemorrhage due to acquired factor VIII inhibitor. An otherwise healthy elderly man developed massive, life-threatening, sublingual bleeding associated with an idiopathic factor VIII inhibitor. The patient was treated wtih cyclophosphamide, steroids, factor VIII concentrates, and repeated plasmapheresis (including three times with NCI-IBM blood-cell separator). Rapid clinical and laboratory improvement occurred, with complete disappearance of the inhibitor. The patient has remained well, without evidence of an inhibitor, for 8 mo. The possible role of each of the therapeutic measures in the disappearance of the inhibitor and the possible pathogenetic mechanism of this disorder are discussed. A high mortality rate and a striking incidence of sublingual hematoma have been observed in cases in the literature.


The effect of heparin on platelet aggregation by common inductors and by ristocetin in congenital bleeding disorders due to factor VIII or fibrinogen defects. Platelet aggregation by various inductors was studied in citrated and heparinized plasma of the following groups of subjects: Normal, hemophilia A, combined factor V and factor VIII deficiency, v. Willeprand's disease and congenital afibrinognemia. The results may be summarized as follows: A-platelet aggregation in citrated plasm 1) platelet aggregation by common inductors ADP, adrenalin and collagen was normal in all groups of subjects but for the patients with congential afibrinogenemia in whom adrenalin induced aggregation was absent or markedly refuced whereas ADP and collagen gave slightly reduced or near normal aggregation curves. 2) platelet aggregation by ristocetin was normal in all groups of subjects but for v. Willebrand's disease in which it was absent. B-platelet aggregation in heparized plasma. 1) platelet aggregation by common inductors resulted to be normal in all groups of subjects except in congenital afibrinogenemia. In this latter case the pattern was still mildly defective but here was an increased aggregation as compared to citrated plasma. These findings have been interpretemmon inductors. 2) platelet aggregation by ristocetin resulted to be absent in all groups of subjects investigated. The possible mechanism of action of the inhibitory effect exercised py heparin with regard to restocetin is discussed.


Acute segmental ischaemic enteritis in Thialand. A disease characterized by segments of ischaemic small intestine has been recognized in norther Thailand over the past decade. The clinical features and appearance of the diseased intestime are described. Most of the patients were treated by surgical resection of the affected bowel. The overall mortality was 14 per cent. Recently, some patients have been successfully treated by non-surgical means. The aetiology of the disease is still unknown.


Locomotion in the cat: basic programmes of movement. Observations in cats of flexion and extension movements of the 4 limbs have led to the conclusion that the different forms of alternative locomotion (e.g. walking, trotting, swimming) and in-phase locomotion (galloping, jumping) result from the interaction of 'programmes' for the coordination of (1) the homologous limbs (pair of hindlimbs or pair of forelimbs) and (2) the homolateral limbs (hind- and forelimb of the same side of the body). The movements of the homologous pairs of limbs are coupled out of phase in alternate locomotion and approximately in phase in the phase form of locomotion. The movements of the homolateral pairs of limbs occur approximately out of phase in the trotting type of coupling and approximately in phase in the pacing type of coupling. Transitions between the different forms of coupling occur abruptly over 1 or 2 steps. Therefore, for each type of coupling (homologous or homolateral) there are two distinct forms or 'programmes' of movement. The hypothesis is advanced that (a) all the characteristic patterns of locomotion in the cat result from different combinations of these 'programmes' of homologous and homolateral limb coupling; (b) the 'programmes' are mutually self reinforcing in the gaits in which the coordination of the movements of the 4 limbs is bilaterally symmetrical; (c) the 'programmes' act in competition in certain gaits which are not bilaterally symmetrical giving rise at times to a changing gait pattern, and (d) the temporary dominance of one 'programme' or another can determine the gait of the particular step.


Different sites of membrane action for tetrodotoxin and lipid-soluble anesthetics. 1. To examine whether different local anesthetics act on different or similar sites in the nerve membrane, the effects of combinations of lipid-soluble anesthetics and tetrodotoxin (TTX) were studied on the compound action potential of the frog sciatic nerve. 2. The nerve-blocking potency of TTX was potentiated 5-fold by benzyl alcohol and 10-fold by lidocaine, at concentrations of these latter two drugs which were just barely anesthetic; no such synergism occurred between saxitoxin (STX) and TTX, their combined effect being merely additive. 3. Simple additive effects also occurred between benzyl alcohol and phenol, benzyl alcohol and urethane, as well as lidocaine with a spirosuccinimide, RAC 109 II. 4. Synergism occurred between STX and 2-aminobenzimidazole. 5. A small but consistent synergism occurred between lidocaine and benzyl alcohol. 6. It is concluded that uncharged anesthetics, tertiary amines, and TTX act via different membrane binding sites which are all closely associated with the Na+ conductance channels.


Evidence for origin of certain childhood acute lymphoblastic leukemias and lymphomas in thymus-derived lymphocytes. Lymphoblasts from children with acute lymphoblastic leukemia (ALL) or malignant lymphoblastic lymphoma were studied using surface markers characteristic of T and B lymphocytes. A B-cell marker, i.e. surface immunoglobulin, was absent in all cases studied. Fouteen of 22 children (64%) had lymphoblasts with one or both markers of T lymphocytes, i.e. receptors for sheep erythrocytes (E) and/or human T-lymphocyte antigen (HTLA) detectable using heterologous antithymocyte sera absorbed with B lymphocytes. In all instances, lymphoblasts which carried E receptors also carried HTLA. However, lymphoblasts in 6 cases carried HTLA but not E receptors. It is possible that ALL may often involve T lymphocytes which are early in differentiation (i.e. prior to development of E receptors) or, alternatively, that E receptors may be lost from T cells following malignant transformation. Thymus enlargement was found only in cases of ALL or lymphoma where T markers were present. Lymphoblasts carried the same markers when examined in various sites and at various times from the same patient.


Electron microscopic observations on the retino-preoptic pathway of Rana temporaria L. Electron microscopic observations confirm the presence of optic terminals of retinal origin in the preoptic area of Rana temporaria. After unilateral section of the optic nerve, degenerating axon terminals were observed among the neurosecretory cells of the preoptic nucleus, both ipsilaterally and contralaterally. The retino-preoptic terminals apparently establish axo-dendritic synapses with non-neurosecretory neurons. Many more degenerating fibres were seen in the ipsilateral preoptic nucleus than in the contralateral nucleus. These are presumed to pass through and to cross in the dorsal posterior part of the optic chiasm.


Scanning electron microscopy of the wall of the third ventricle of the brain of Rana temporaria. III. Electron microscopy of the ventricular surface of the median eminence. From combined scanning and transmission electron microscopic studies it is concluded (1) that all surface specializations of the median eminence of Rana temporaria are protrusions of ependymal cells; (2) that the microvillus-like structures of the median eminence are transient differentiations of the ependymal cell surface, associated with the activity of the ependymal cells.


Determinants of reversible asynergy. The native coronary circulation. To evaluate the influence of the native coronary circulation on the reversibility of asynergy, ventriculograms before and after sublingual nitroglycerin were performed in 51 patients with coronary artery disease and asynergy. The severity of stenotic lesions and caliber of the distal coronary vessels were determined by comparison with external catheter tip diameter corrected for magnification. Of 42 asynergic zones associated with larger than or equal to 90% proximal coronary occlusion, 27 (64%) were akinetic or dyskinetic while only 11 of 38 zones (29%) with less than 90% occlusion showed akinesis (P less than 0.005). Twenty-six of the 38 asynergic zones (69%) with less than 90% occlusion were reversible in contrast to 19 of the 42 zones (45%) with larger than or equal to 90% occlusion (P less than 0.05). Coronary collaterals were observed in 23 of 42 (55%) zones with larger than or equal to 90% occlusion in contrast to only 11 of 38 zones (29%) with less than 90% occlusion (P less than 0.05). Of the zones with both greater than or equal to 90% occlusion and collaterals, 74% were reversible, in contrast to only 11% without collaterals (P less than 0.001). Of the asynergic zones without collaterals, 63% with less than 90% occlusion were reversible in contrast to only 11% with larger than or equal to 90% occlusion (P less than 0.001). Pathologic Q waves were associated with 24 of 42 zones (57%) with larger than or equal to 90% occlusion compared to only nine of the 38 zones (24%) with less than 90% occlusion (P less than 0.01). The presence of Q waves was associated with a significant decrease in the incidence of reversibility regardless of the degree of coronary occlusion. Excluding the asynergic zones with either collaterals or Q waves, 79% with less than 90% occlusion were reversible in contrast to only 37% with larger than or equal to 90% coronary occlusion (P less than 0.05). In contrast, the caliber of the distal vessel could not be correlated with either the severity of asynergy or the presence of collaterals and was similar in both reversible and irreversible asynergic zones. In summary, larger than or equal to 90% proximal stenosis is associated with severe asynergy which is less likely to be reversible compared to asynergy associated with less than 90% coronary occlusion. In the presence of larger than or equal to 90% occlusion, coronary collaterals are associated with a significantly higher incidence of reversible asynergy and thus appear to serve a protective function. However, the caliber of the distal vessel per se does not effect the severity or reversibility of asynergy.


Transbronchial lung biopsy via the fiberoptic bronchoscope. Experience with 107 consecutive cases and comparison with bronchial brushing. Transbronchial lung biopsy (TBB) was performed during fiberoptic bronchoscopy under fluoroscopic guidance in 107 patients. TBB was diagnostic in 17 of 21 (81 percent) cases with localized malignant lesions greater than 4 cm in diameter and bronchial brushing was positive in 12 (57 percent). TBB was diagnostic in 14 of 24 (58 percent) cases with localized malignant lesions equal to or less than 4 cm in diameter, whereas brush biopsy was positive in 7 (29 percent). In 13 cases with suspected Pneumocystis carinii pneumonia, the combination of TBB and brush biopsy was diagnostic of P carinii pneumonia in all 11 patients; TBB was diagnostic in 10 and brush biopsy diagnostic in 5. Cytomegalovirus pneumonitis was diagnosed in the other two cases. In 20 patients with localized infiltrates or nodules, a TBB diagnosis of acute or chronic inflammation excluded malignancy in 15 of 18 cases (follow-up 3 to 24 months) and tuberculosis was diagnosed in 2. Malignancy was found in one patient with acute inflammation on TBB. TBB accurately diagnosed 23 of 29 (79 percent) cases of diffuse lung disease. Following TBB, one patient had pneumothorax and nine patients had hemoptysis not requiring treatment. TBB and bronchial brushing via the flexible fiberoptic bronchoscope offer good diagnostic accuracy and a low complication rate in both diffuse and localized lung diseases.


Spontaneous graft closure in anomalous origin of the left coronary artery. Several reports of successful correction of anomalous origin of the left coronary artery from the pulmonary artery utilizing a graft to the ascending aorta have demonstrated the feasibility of this procedure. The patient described in this report developed proved delayed occlusion of the saphenous vein graft with a fatal outcome. This was a result of intimal fibrous hyperplasia identical to that seen in adults following the placement of the saphenous vein bypass graft for treatment of atherosclerotic coronary disease.


[Clinical and prognostic significance of antinuclear factors during D-penicillamine treatment (author's transl)]. Antinuclear factors were demonstrated during D-penicillamine treatment in patients with rheumatiod arthritis (72) and scleroderma (7). While in 13 antinuclear factors were demonstrable already before treatment was started, they occurred in the course of treatment in another 34. D-penicillamine administration had to be discontinued in 18 because of severe side effects, with a marked correlation between  their severity and the presence of antinuclear factors. Side effects requiring stoppage of the drug were especially common in patients who had antinuclear factors even before onset of treatment. The severest complication of D-penincillamine treatment, renal damage, was found only in patients with antinuclear factors, in this instance antibodies against denatured DNA. The following therapeutic consequences should be drawn from these findings: (1) D-penicillamine should, if possible, not be given to any patient with antinuclear factors; (2) in patients in whom antinuclear factors occur in the course of D-penicillamine treatment but its administration is to be continued, blood picture, renal functions and urine must be regularly tested.


[Clinical course of typhoid in the Heidelberg region (author's transl)]. In the course of a typhoid epidemic during the autumn of 1974 in the Heidelberg region 74 persons were treated in hospital. Chloramphenicol was give to 45, ampicillin to 19. The former, in daily doses of 2.0 g, gave worse results if given for only two instead of three weeks. In comparison, ampicillin was less effective. A second course of treatment became necessary in 13 patients, with trimethoprim-sulphamethoxazole (Bactrim) being succesful in all, although the follow-up period is still too short for definitive results. Three complications occurred: one case of massive bleeding from the gut requiring operation and followed some weeks later by a HBS-antigen-negative hepatitis; one case of typhoma (several weeks after the end of antibiotic treatment), requiring operative removal; a case of febrile abortion in the second month of pregnancy.


The amino-acid sequence of the most acidic major parvalbumin from frog muscle. The primary structure of the most acidic (pI - 4.50) of the two major parvalbumins from frog (Rana esculenta) has been determined from a study of its trypsic peptides and of overlapping peptides generated by limited trypsic digestion, chymotrypsic digestion and N-bromosuccinimide cleavage of the protein. The amino acid sequence so obtained is considered in comparison with those known for other parvalbumins and for rabbit troponin-C.


Dilantin-calcium interaction and active Na transport in frog skin. The effects of diphenylhydantoin (DPH) and calcium were simultaneously studied on solute transport in frog skin. In the presence of external Ca, DPH elicited a significant increase in active Na transport; in its absence, DPH had no significant effect on active transport. DPH increased passive permeability, but this action was independent of Ca. It appears that the increase in active transport elicited by DPH is due to a specific interaction with Ca at the transport site.


Cell membranes as sources of granulocyte colony stimulating activities. Subcellular localization of factors associated with stimulation of granulocyte progenitors (CFU-c) to form colonies in vitro was investigated. Human peripheral blood leukocytes were fractionated into three subcellular fractions. After solubilization with sodium dodecyl sulfate, fractions were assessed for ability to stimulate colony formation by CFU-c. The fraction containing cell membranes proved to be the richest source of stimulating factors. Apparent molecular weights for these factors as determined by gel filtration were equivalent to those obtained for factors from purified peripheral leukocyte conditioned medium. The results suggest that at least some granulocyte colony-stimulating factors should be regarded as membrane-active agents.


Serum potentiation of granulocyte and macrophage colony formation in vitro. Addition of serum or plasma from many species increased the number and size of neutrophilic granulocyte and macrophage colonies formed in vitro by mouse bone marrow cells after stimulation by a variety of preparations containing colony stimulating factor (CSF). The active factor in serum was heat-labile, non-dialyzable and eluted from G-100 columns in the serum albumin region. Serum non-selectively affected all cells potentially capable of forming granulocytic and macrophage colonies but did not potentiate the formation of eosinophilic or megakaryocytic colonies. The potentiating activity of mouse serum was increased by prior injection of the mice with endotoxin but was not affected by prior whole-body irradiation. Colony potentiating activity in plasma from patients with myeloid of lymphoid leukemia did not differ from that of normal human plasma.


The influence of mouse sera on colony formation and on the production of colony stimulating factor in vitro. Results of studies on the in vitro production and release of colony stimulating factor (CSF), by bone associated cells cultivated in the presence or absence of mouse sera, are presented. Tissues from three strains of mice were shown to be equally effective in the production of CSF in vitro. Priming of the tissue donors by endotoxin 4 hours previously, or by irradiation 1, 7 or 15 days previously, significantly increased the CSF production. The addition of serum from normal mice to the culture medium had no effect on CSF production by tissue from normal mice but regularly inhibited CSF production by bones from irradiated animals. The inhibition was not due to the toxic effect of the sera on the CSF producing cells and was more pronounced in the cultures of NMRI than in the C57BL tissues. Some inhibition was also observed in cultures of tissue from endotoxin-primed NMRI mice. Extraction of the sera with ether removed the inhibitory material. The sera which inhibited the CSF production in vitro also inhibited the growth of granulocytic and macrophage colonies in soft agar cultures but did not inhibit the growth and DNA synthesis of cultured L-cells.


Enhancement of chicken hemagglutinin response by thymocytes. Antigen responsive cells, detectable with the cell transfer antibody assay, are abundant in the blood and spleen of chickens after immunization. In experiments where serial dilutions of donor immunocyte-antigen mixture were cultured in host embryos, each two-fold dilution of the mixture resulted in approximately a four-fold reduction in hemagglutinin output. When higher than normal concentrations of donor buffy coat were used, however, serial dilutions resulted in antibody output that was more directly proportional. The overall dose-response curve of relative donor cell number to antibody production appears to be biphasic, with a slope of a first order reaction at higher concentrations, and a steeper slope at lower concentrations of donor cells. The hemagglutinin response in the adoptive cell transfer system was enhanced when autologous thymocytes were added to the immunocyte-antigen mixture. Data from the kinetic and thymocyte supplement experiments taken together strongly indicate that the mouse erythrocyte antigen responsive cells of the chicken, like their mammalian counterpart the antigen-reactive units, involve the cooperation of T and B type lymphocytes.


Relationships between H-2 Region and surface characteristics of mouse spleen cells. The purpose of this investigation was to study whether allelic variations at the H-2 region which induce differences in the peptide composition of some glycoproteins of cell surface, also induce variations in electrical charges at the cell membrane. The surface charge was assessed by the electrophoretic mobility of the cell. Tests with different congenic strains were undertaken on spleen cells which were assumed to contain two major lymphocyte populations with different surface charge. The data show that the mean electrophoretic mobility of the low mobility lymphocytes was independent of the strain. By contrast, some H-2 haplotypes give 8% difference in surface charge of high mobility lymphocytes. These results suggest that the H-2 region plays a role in the regulation of ionized groups of cell surface in some but not all populations of spleen lymphocytes. The high and low mobility lymphocyte populations are assumed to represent T and B lymphocytes, respectively. The proportions of T lymphocytes obtained with electrophoretic marker were higher than those observed in other strains with currently available markers. It is suggested that the high mobility population contains T and "null" lymphocytes. Moreover, the data show that the H-2 region controls the relative proportions of T (or T and "null") and B lymphocytes, the H-2s specificity inducing a B/T ratio different from that induced by other specificities.


Haemochromatosis, active chronic hepatitis, and familial IgA deficiency. A man aged 38 years developed chronic liver disease with characteristics of both haemochromatosis and active chronic hepatitis. Two of his relatives were found to have selective IgA deficiency. It is suggested that this case provides evidence that haemochromatosis may initiate active chronic hepatitis in a genetically susceptible subject.


Prevention by glucagon of experimental stress induced hemorrhagic gastritis. Glucagon was effective in decreasing acute gastric mucosal lesions, blood loss and gastric acid secretion in the cold restraint stressed rat.


A histochemical study on the central effect of monoamine precursors. A histochemical investigation has been made on the central monoamine neurons after the administration of monoamine precursors and peripheral decarboxylase inhibitor in experimental animals and the results are summarized as below: (1) Catecholamine fluorescence in the brain reduces and yellowish indoleamine fluorescence manifests after the treatment with 5HTP alone and Ro4-4602 plus 5HTP. These findings support the view that catecholamine is likely to be replaced by indoleamine. (2) Ro4-4602 does not cause directly any change in central catecholamine neurons except in median eminence where the inhibitor causes a decrease in dopamine fluorescence. The findings indicate that the inhibitor may act in the hypothalamus as same manner as in the peripheries, because the hypothalamus is presumably lacking of blood brain barrier. (3) L-Dopa alone and L-Dopa plus Ro4-4602 enhance dopamine fluorescence in the substantia nigra and the median eminence, especially an increase in the extra-neuronal fluorescence can be seen in the latter. This indicates the releasing of amines from cell body into extraneuronal spaces.


Immunosuppressor T cells in tumor bearing host. Thymus or spleen cells of tumor bearing animals (TBA) (methylcholanthrene induced sarcoma bearing A/Jax mice) were shown to possess immunosuppressor cells regulating the immune response to the tumor in syngeneic animals. The immunosuppressive activity of these cells of TBA was totally abolished by the in vitro treatment of anti-theta serum and complement. Soluble factor(s) with identical suppressive activity was isolated from the thymus cells of TBA and its molecular size was estimated to be lower than that of serum albumin in terms of its elution behaviour on gel filtration through a Sephadex G-200 column. These immunosuppressor T cells or their soluble factor(s) may be largely responsible for the growth of antigenic tumors.


In Vitro studies on human B and T cell purified populations. Stimulation by mitogens and allogeneic cells, and quantitative binding of phytomitogens. The property of T cells to form rosettes with sheep red blood cells has been used to separate peripheral blood lymphocytes into purified T- and B-cell suspensions after density gradient centrifugation. A study of lymphocyte markers has shown that 2-6 per cent of E rosettes only were recovered in the B cell-enriched population. Lymphocyte stimulation in vitro was obtained with PHA, con A and PWM in unseparated and T-cell populations, but never in B-cell suspensions. Experiments of recombination between the two purified fractions have demonstrated that 10% of T cells added to B cells were able to induce a response to PHA and PWM. Otherwise, only T cells responded to allogenic stimulation. Lastly, B and T cells seemed to bind qualitatively and quantitatively the same mitogens on their membranes.


The activity of T and B lymphocytes in immunity and tolerance to the lymphocytic choriomeningitis virus in mice. Treatment with anti-theta serum and the Wigzell column technique for cell separation was employed to study the separate functions of the B and T lymphocytes in the late states of immunity to the LCM virus in mice. The cell preparations examined were mixtures of spleen and lymph node cells from immune mice. The results revealed that the anti-viral effect of such cells after transfer to virus carriers was unimpaired in T cell-enriched and B cell-deprived cell preparations. The anti-viral effect was also retained in cell preparations deprived so much of B cells that no antibody was produced in the virus carrier mice receiving transplants of these cells. The results strongly indicate that the anti-viral effect of late immune cells is not only T cell-dependent but that it is also mediated solely by T cells and, moreover, that antibodies have no or very little influence on the virus elimination. The observation that antibody production could be caused neither by column-passed cells nor by anti-theta serum-treated cells, but was obtained by mixtures of these cells, demonstrates that co-operation between T and B cells is crucial for the LCM antibody response. Accordingly, the convincing demonstration of the absence in the persistent virus carriers of cells which, in respect of antibody production, are able to co-operate either with column-passed or with anti-theta serum-treated immune cells, implies that such animals are extremely deficient as regards immune function of both B and T LCM-primed lymphocytes.


Epicutaneous induction of hyporeactivity in contact sensitization. Demonstration of suppressor cells induced by contact with 2,4-dinitrothiocyanatebenzene. Partial unresponsiveness to 2,4-dinitrofluorobenzene (DNFB) was produced by the epicutaneous application of 2 per cent 2,4-dinitrothiocyanatebenzene (DNTB) to the dorsum of the ear, 14 and 7 days before sensitization. This state of partial unresponsiveness could be broken by treatment with 300 mg/kg cyclophosphamide (CY) 3 days before the sensitization dose of DNFB. There was a significant increase in the uptake of 125I-labelled iododeoxyuridine in the draining lymph node, 6 days after the application of 2 per cent DNTB and in the contralateral nodes at 4 and 6 days. The state of partial unresponsiveness to DNFB was not associated with a decrease in T-cell proliferation in the draining lymph nodes. The generation of suppressor cells, capable of reacting in the periphery, was demonstrated by passive cell transfer into recipients sensitized to DNFB and depleted of normally produced suppressor cells by pretreatment with CY. These cells were found in spleen and peritoneal exudates of animals treated by the epicutaneous application of 2 per cent DNTB 14 and 7 days previously. It is suggested that the suppressor cells induced by contact with DNTB act by competition with effector cells at the periphery.


Investigation of NZB mice born to BALB/c immunized against NZB allotype. Allotype suppression is held to be an example of T-cell suppressor function. Failure of generalized suppressor T-cell function is considered to be responsible for the chronic and progressive disease of the NZB. The possible association of allotype suppression and autoimmune disease has been investigated here in a group of NZB mice, transplanted and born from BALB/c recipients whilst producing anti-NZB allotype. In this situation it was anticipated that if allotype suppression should fail this would coincide with the development of autoimmune disease; both processes reflecting failure of T-cell suppressor function. However, in spite of the fact that allotype suppression was not achieved, certain observations seem important. The fact that NZB born the BALB/c were in no way different from normally derived NZB confirms that the cause of the NZB disease is established prior to the stage of implantation. Maternal influence at or beyond this stage appears of little consequence in terms of effecting the development or progression of the NZB disease. The failure to induce allotype suppression in the NZB is important. In this context this strain appears to be no different from the majority of other strains that have been tested as homozygotes. The suppression obtained with (BALB/c X SJL/J)F1 hybrids in which chronic suppression has been achieved therefore seems to represent an exceptional situation and this suggests that it is perhaps unwise to base general assumptions as to the universal adaptability of T-cell control mechanisms upon unusual findings. It must be remembered that only homozygotes were examined here. Earlier attempts to induce allotype suppression in homozygotes also failed, even in the SJL/J. Since allotype suppression has only been demonstrated in heterozygote mice there is a distinct possibility that this phenomenon never occurs in the homozygote situation. The significance of this possibility is discussed.


Presence and kinetics of sensitized cells in different tissue compartments. The question was studied as to whether lymph nodes (LN), spleen (SP) and bone marrow (BM) contain different amounts of sensitized cells at various times after one or repeated immunizations. The following model was used: C57 mice were injected i.p. once, twice or three times with C3H cells. Between 3 and 20 days after the last immunization, LN, SP and BM cell suspensions were tested in vivo for their local cutaneous lymphocyte transfer reactivity in C3H mice and in vitro for their lymphocytotoxicity (partially after preincubation with anti-theta serum). In both test systems similar observations were made: after one sensitization, reactive cells were demonstrated first in the LN and then in the SP, but only during the next few days; BM cell reactivity appeared later. In the first few days after two sensitizations SP cells reacted strongly whereas LN and BM cells reacted weakly; later BM cell reactivity increased while the others decreased. After three sensitizations reactivity of the SP cells predominated during the entire period of observations; the BM cells showed less, the LN cells no reactivity. These findings suggest dependance of T cell-mediated reactivity on (1) the origin of the cells from LN, SP or BM, (2) the time of the harvesting from these tissues, and (3) the degree of sensitization.


Human tumor--lymphocyte interaction in vitro: blastogenesis correlated to detectable immunoglobulin in the biopsy. Binding of radioiodinated anti-immunoglobulin (Ig) reagent assayed by acid elutable radioactivity was shown in 18 of 44 cell suspensions (41%) prepared from surgical specimens of human tumors. Aliquots of these biopsies were admixed to autologous lymphocytes and in 13 cases they induced stimulation of DNA synthesis. In only one of these 13 cases was the anti-Ig reagent bound, while among the 26 biopsies with low or no binding capacity 12 (46%) were stimulatory, indicating that immunoglobulin-containing biopsies are not stimulatory. Experiments on 32 lymphocyte preparations from different lymphoid organs suggest that the immunoglobulin detected in the tumor-cell suspension is not derived from the infiltrating lymphoid cells.


Comparison of 51Cr release and microcytotoxicity assays against human melanoma cells. Comparisons of the cytotoxic activity of mononuclear cells from venous blood of human subjects against human melanoma cell lines have been made by means of the 51Cr release assay (CRA) and the microcytotoxicity assay (MCA) of Takasugi and Klein. The relative effectiveness of the two assays in the detection of killing by various sub-populations of effector cells was studied after depletion of monocytes and separation of the blood monocluear cells by SRBC receptor and Fc receptor resetting techniques. Overall, the results showed good correlation. Most differences were the result of low values recorded with the CRA when effector cells were obtained from Hypaque-Ficoll--separated populations which had not been further fractionated. These differences were largely removed when monocytes were depleted by glass absorption and may reflect interference with the CRA by monocytes. No consistent differences were found between the assays in the detection of killing by subfractions enriched for T cells or B cells in this tumour system. The MCA appears to offer the advantage of greater sensitivity and possibly detection of a wider range of cytotoxic mechanisms but the CRA has a number of advantages of a technical nature which make it more applicable to routine assays in clinical studies.


A study of the oncogenicity of adenovirus type 2 transformed rat embryo cells. Rat embryo cells were transformed by adenovirus type 2 at different multiplicities of infection. The oncogenic potential of the resulting transformed lines varied considerably but a relationship was found to exist between transforming virus dose and oncogenicity of the resulting transformed lines. Possible reasons for the considerable variation in oncogenicity of the transformed lines are discussed, giving particular consideration to cellular antigenicity and recent molecular biology studies of adenovirus type 2 transformed cell lines.


Specific anti-tumor responses of cultured immune spleen cells III. Further characterization of cells which synthesize factors with blocking and antiserum-dependent cellular cytotoxic (ADC) activities. Spleen cells from BALB/c mice bearing syngeneic, methylcholanthrene-induced sarcomas were cultured in vitro. In agreement with previous observations, two tumor-specific activities could be demonstrated in the culture supernatants: the supernatants suppressed (blocked) specific lymph-node cell-mediated cytotoxicity directed against the respective neoplasm and they induced specific antibody-dependent-cellular cytotoxicity (ADC) mediated by lymph-node cells from non-immune mice. In the present study, the effects of specific depletion or enrichment of different celltypes on the ability to synthesize the blocking and ADC activities was explored. Spleen cells passed through columns of Sephadex G-10, which removed plasma cells and macrophages, no longer synthesized factors with ADC activity though synthesis of blocking factors continued. However, lysis of Thy-1-positive cells with antiserum and complement did abolish the synthesis of blocking factors. Spleen cells from tumor-bearing mice were then enriched for Thy-1-positive cells by selective removal of macrophages and plasma cells on G-10 columns and of immunoglobulin-bearing cells on anti-mouse immunoglobulin affinity columns. This T-cell-enriched population synthesized blocking factors and restored blocking factor synthesis in cultures depleted of Thy-1-positive cells. Simiarly enriched spleen cells from normal control mice did not synthesize tumor-specific blocking factors but partially restored synthesis of blocking factor in tumor-bearer spleen cultures depleted of Thy-1-positive cells.


Antibody-dependent cell-mediated cytotoxicity for human monolayer target cells bearing blood group and transplantation antigens and for melanoma cells. A sensitive assay for antibody-dependent cell-mediated cytotoxicity (ADCMC) was developed utilizing serum from a patient with gestational choriocarcinoma and the 3H-proline microcytotoxicity test for detection fo destruction of monolayer target cells. Conditions for optimal expression of ADCMC were investigated using serum from this patient and skin fibroblasts from her husband and daugther as target cells with semi-purified blood leukocytes from normal donors as effector cells. Factors critical for optimal expression of ADCMC in this assay are the selection of effector cell donors possessing high levels of activity in the presence of serum with known lymphocyte-dependent antibody. (LDA), the effector cell preparative technique, and  incubation for up to 40 h. Tris-NH4Cl lysis of red blood cells was found significantly to reduce effector cell activity. Under optimal conditions, the LDA titer of this patient's serum was greater than 10(-4). The sensitivity of the assay was confirmed by the detection of LDA activity of an anti-blood-group antibody at dilutions not demonstrating hemagglutination, and by the induction of ADCMC for blood group A antigen-bearing target cells by normal sera of B and O blood groups. In a preliminary study of sera from 16 melanoma patients on the corresponding antologous tumor target cells, four had significant LDA. Further studies are under way to determine specificty, incidence, and relationship of LDA-positive autologous sera to course of disease.


The relationship between tumorigenicity, growth in agar and fibrinolytic activity in a line of human osteosarcoma cells. The TE-85 osteosarcoma cell line has several of the in vitro properties of malignant cells, including colony formation in agar, but has low extracellular fibrinolytic activity and no capacity to form tumors in ATS-treated hamsters. Some TE-85 cells clones (clones 2, 4 and 6) have increased fibrinolytic activity but do not form tumors in hamsters. TE-85 cells infected with mammalian transformation-defective viruses have low (FeLV) or increased (RD-114 virus) levels of fibrinolytic activity and do not form tumors in hamsters. TE-85 cell either nonproductively infected with Ki-MSV or productively infected with M-MSV (RD-114), have fibrinolytic activity and can form tumors in hamsters. The MSV gene(s) but not colony formation in agar or extracellular fibrinolytic activity appears to be capable of rendering TE-85 cells tumorigenic in ATS-treated hamsters.


Rejection of tumor cells in vitro: a T-cell-mediated reaction. Cytoxic lymphoblasts (LBC) were produced by the co-cultivation of guinea-pig lymphoid cells from the thymus, lymph nodes, spleen, bone marrow and blood with syngeneic methylcholanthrene-induced sarcoma (MC-D) cells. The cytotoxic reaction was inhibited by pretreatment of the lymphoid cell populations with a rabbit antiserum specific for guinea-pig thymus-derived (T) lymphocytes. Supernatants of LBC inhibited the migration of peritoneal exudate cells, but did not contain lymphotoxin or a skin reactive factor. It is concluded that the cytotoxic cells are of T-cell origin.


Reduction in T-lymphocytes forming active rosettes in chronic marijuana smokers. Rosettes formed by peripheral T- and B-lymphocytes were studied in 23 normal controls and 23 marijuana smokers. The mean percentage of cells forming B type rosettes were normal in both groups. The percentage of T cells forming rosettes was lower in the marijuana smokers, and 39% had values below 2 standard deviations of the mean of the normal control group. These observations suggest some alteration in some T cells in some marijuana smokers.


Mechanism of gap-filling during postreplication repair of ultraviolet damage in Haemophilus influenzae. Deoxyribonucleic acid (DNA), pulse labeled after ultraviolet irradiation of excision-defective mutants of Haemophilus influenzae, is of lower single strand molecular weight than that of unirradiated cells but approaches the size of DNA from unirradiated cells upon further incubation in growth medium. This gap-filling process is controlled by the rec-1 gene. Gap-filling occurs normally in a temperature-sensitive DNA synthesis mutant at the restrictive temperature showing that normal semiconservative DNA synthesis is not necessary for gap-filling. To test for recombinational events after irradiation, the DNA synthesized after irradiation was radioactively labeled for a short time in medium containing 5-bromodeoxyuridine followed by incubation for various times in non-radioactive, 5-bromodeoxyuridine-containing medium. The DNA was denatured and analyzed isopycnically. The labeled DNA was initially "heavy," but later shifted toward lighter densities. This shift occurred in the temperature-sensitive DNA synthesis mutant at the restrictive temperature and in the recombination-defective mutant rec-2, but was not seen in the rec-1 mutant. The density shift can be interpreted as evidence that rather extensive exchanges occurred between parental DNA and the DNA made after irradiation. These results suggest that such exchanges are important for gap-filling in H. influenzae.


Mechanism of action of ethanolamine ammonia-lyase, an adenosylcobalamin-dependent enzyme. Proton nuclear magnetic resonance studies of the binding of adenine nucleosides and substrate to ethanolamine ammonia-lyase. Proton NMR spectroscopy was used to study the binding of adenosine, 5'-deoxyadenosine, adenine, and ethanolamine to the adenosylcobalamin-dependent enzyme ethanolamine ammonia-lyase. Broadening of ligand resonances in the presence of ethanolamine ammonia-lyase indicated that adenosine, 5'-deoxyadenosine, and ethanolamine all formed complexes with the enzyme (KD(mM) = 3.5, 3.0, and 2.5 respectively). The methyl group of enzyme-bound 5'-deoxyadenosine rotated at a rate exceeding 10(7) revolutions/s. Adenine did not appear to bind to the enzyme. Rates of dissociation of nucleosides from the enzyme were fast on the NMR time scale, precluding measurements of rate constants for the binding reaction. The departure of ethanolamine was slow, however, permitting their determination. The values for these rate constants were: k1 = 4.4 times 10(5) M-1 S-1; k-1 = 1.1 times 10(3) S-1. Addition of 1 mol of cyanocobalamin/mol of active sites led to narrowing of the enzyme-broadened ligand resonances. With 5'-deoxyadenosine, linewidths still exceeded those of the free ligand, indicating that binding to enzyme was weakened but not abolished. The KD for this nucleoside in the presence of CNCbl was 8.0 mM. With ethanolamine and adenosine, however, linewidths reverted to values characteristic of the unbound ligand, indicating either that CNCbl greatly lowered the rate of dissociation of the ligand or displaced the ligand from the enzyme. A decision between these two possibilities could not be made from the data at hand, although analogy with the situation obtaining with 5'-deoxyadenosine suggests that adenosine is displaced from the enzyme by CNCbl. 5'-Deoxyadenosine inhibited catalytic activity of the enzyme, competing with adenosylcobalamin (Ki = 2.7 mM). Adenosine had no effect, despite NMR evidence indicating that it formed a complex with free enzyme. These experiments showed that ethanolamine ammonia-lyase possesses binding sites for adenine nucleosides, a class of compounds chemically related to the Cobeta-ligand of the cofactor, as well as for ethanolamine. Binding to the enzyme has now been demonstrated for all three categories of low molecular weight compounds thought to be involved in the reaction; namely, substrate (ethanolamine), corrin, and adenine nucleoside.


Nucleocytoplasmic transport and distribution of an amino acid, in situ. Ultra-low temperature techniques (microdissection and autoradiography) were used to study the nucleocytoplasmic distribution and transport of alpha-aminoisobutyric acid (AIB) in an amino acid-accumulating cell. In amphibiam oocytes incubated in AIB, the nuclear concentration of this non-metabolizable  amino acid exceeds the cytoplasmic concentration by 45%, remaining constant both over time and variation in substrate concentration. The kinetics of uptake suggest that this nucleo-cytoplasmic asymmetry arises from solubility differences between the 2 compartments, and that the nuclear envelope plays a negligible role in amino acid transport. A solute exclusion model is offered to explain the nucleocytoplasmic asymmetry.


Detection of adenovirus in patient specimens by indirect immune electron microscopy. Immune electron microscopic procedures for the detection of adenovirus type 7 directly in throat swab specimens from patients are described. Nineteen of 25 throat swab samples, known to be positive for adenovirus type 7 by isolation of virus from tissue culture, were shown to contain aggregates of adenovirions coated with antibody. Sensitivity tests of the immune electron microscopic method showed that as few as 16 to 32 tissue culture infective doses of virus could be detected by the direct immune electron microscopic technique. It was also demonstrated that aggregation of virus-antibody complexes could be further enhanced by use of anti-immunoglobulin G sera (indirect immune electron microscopic procedures). These results demonstrate that examination of patient specimen by immune electron microscopic procedures is a feasible and rapid method for adenovirus detection and suggest that it could be applied as a routine laboratory procedure for the diagnosis of other virus infections.


[Leiomyoma of Meckel's diverticulum revealed by a digestive hemorrhage. Preoperative diagnosis by selective arteriography]. The authors report a case revealed by low digestive hemorrhage, and in the light of cases in the literature, discuss the various problems due to leiomyomas in Meckel's diverticulum. They emphasize particularly the risk of late diagnosis at the stage of leiomyosarcoma. As it is impossible to say afterwards whether a muscular tumour of the digestive tract is benign or not, it is important to make an early diagnosis. Arteriography is one of the best means.


Obligately anaerobic strains of Corynebacterium vaginale (Haemophilus vaginalis). Six obligately anaerobic strains of Corynebacterium vaginale (Haemophilus vaginalis) have been isolated and their characteristics studied. The reactions of the anaerobic strains, as well as of facultative strains tested under anaerobic conditions, are similar to the reactions previously reported for this species.


Problems in the detection of carriers of haemophilia A. Factor VIII activity and factor VIII related--or Willebrand--antigen were studied in 49 known carriers of haemophilia A and 31 normal women, and the data were analysed by four statistical approaches. Sixteen per cent of normals and 18% of carriers were misclassified, overlapping with the other group. Although the percentage of carriers detected is higher when taking into account the results of both biological and immunological factor VIII, it is lower than others recently reported, and the discrepancies between the results obtained are discussed.


Unmyelinated preganglionic fibers in frog ventral roots. It has been suggested that the preganglionic visceral fibers in the frog are unmyelinated. To obtain further evidence for this suggestion: (1) the percentages of unmyelinated fibers and the myelinated fiber histograms were done for all frog ventral roots, (2) the number of unmyelinated fibers were counted on either side of a chronic section of the seventh ventral root and (3) compound action potentials were obtained for roots 7-10. The results are that slightly more than 80% of the unmyelinated fibers in the seventh ventral root arise in the spinal cord and are presumably unmyelinated preganglionic efferents. Furthermore, a C fiber volley could be followed from the seventh ventral root into caudal parts of the sympathetic chain. Thus this data provides confirmation for the idea that many frog preganglionic fibers are unmyelinated. This study also shows that 24% of the ventral root axons in the frog are unmyelinated and that, on the basis of percentages of unmyelinated fibers and myelinated fiber histograms, there are five categories of frog ventral roots.


An investigation of the family background of acute Haemophilus infections of children. Nose and throat swabs, for culture of Haemophilus influenza type b, and blood samples, for measurement of antibodies specific for that serotype, were collected from members of 28 families from which children had been admitted to hospital with acute H. influenzae type b infections (mainly meningitis or epiglottitis). The patients with meningitis were younger than those with epiglottitis and had more siblings, with a marked predominance of sisters. Investigations within a few days of admission of the affected children to hospital detected carriers of H. influenzae type b (19 altogether) in 13 of the 28 families, including 9 of the 13 families with 3 or more children. Members with raised antibody titres for H. influenzae type b (suggesting the presence of the organism for at least a few weeks) were found in 17 of the 25 families from which blood samples were obtained, including all 11 families with 3 or more children. Most of the patients probably acquired their infections from within their own families, and siblings under 11 years old were of predominant importance both as carriers and as potential sources of the patients' infections. Persistence of the organism within families for up to 6 months was demonstrated. Possible reasons for the difference in age-incidence between haemophilus meningitis and epiglottitis and for the occurrence of the former in babies with older sisters are suggested, and also a possible connection between the results of this survey and the likely value of immunization against H. influenzae type b.


Regulatory substances produced by lymphocytes. II. Lymphotoxin in the rat. Rat lymphotoxin (LT), a soluble mediator which kills L cells (mouse fibroblasts) within 48 hr, was partially purified by conventional protein fractionation procedures from the culture supernatants of rat lymph node cells stimulated with ovalbumin in vivo and rechallenged with the same antigen in vitro. It appeared, from the elution pattern of DEAE-cellulose chromatography, to be a neutral or slightly basic protein and was inactivated at 60 degrees C. Its molecular weight was estimated by Sephadex gel filtration to be approximately 9 times 10(4) daltons. Partially purified LT, in titration studies, showed single hit kinetics. It also inhibited L cell proliferation at low concentration without significant killing effect. The heat inactivation curves of both cytotoxic and proliferation inhibitory activities were the same, suggesting that these two activities may be caused by the same molecule. However, it had almost no detectable effect on the DNA synthesis of rat lymph node cells stimulated with phytohemagglutinin (PHA), suggesting that lymphocytes, at least those which respond to HA, are rather resistant to LT. It is suggested that activities described as "proliferation inhibitory factor" and "cloning inhibitory factor" by others may represent the activity of dilute LT. The "inhibitor of DNA synthesis" is distinct in molecular character, range of target cells, and target cell kinetics from LT.


An analysis of the defective response of CBA/N mice to T-dependent antigens. CBA/N mice fail to respond to T-independent antigens, and give a diminished response to T-dependent antigens. The cellular basis of the diminished response to T-dependent antigens in this strain was analyzed by means of adoptive transfers of various primed cell populations along with cells from the immunologically normal, histocompatible strain CBA/CaJ. CBA/N mice behaved normally as recipients and had normal levels of spleen helper cell activity. However, CBA/N spleen cells, used as a B cell source, produced only 56% as much antibody as CBA/CaJ cells. Since CBA/N mice have a relative frequency of immunoglobulin-bearing lymphocytes roughly half that of normal mice, these results suggest that CBA/N mice have a quantitative defect in spleen B cells responsive to T-dependent antigens.


Antigenic analysis of Chlamydiae by two-dimensional immunoelectrophoresis. I. Antigenic heterogeneity between C. trachomatis and C. psittaci. Tw0-dimensional immunoelectrophoresis was used for the antigenic analysis of Chlamydia. Antigens of chlamydiae were found to be successfully solubilized with the nonionic detergent Triton X-100. Nineteen and 16 distinct components were observed for Chlamydia trachomatis strain L2/434/Bu and C. psittaci strain Meningopneumonitis/Cal-10, respectively. The triton-solubilized antigens of chlamydiae differ strikingly between species since only one strong reciprocal cross-reacting antigen was observed between 434 and Cal-10. However, the antigens of Trachoma strain K/UW-31/Cx and LGV strain 434 were found to be very similar.


Triggering mechanism of B lymphocytes. I. Effect of anti-immunoglobulin and enhancing soluble factor on differentiation and proliferation of B cells. Stimulation of rabbit mesenteric lymphocytes with anti-immunoglobulin antibody (anti-Ig) for 24 hr (first stage of culture) followed by the addition of enhancing soluble factor (second stage of culture) induced the formation of IgG. The presence of mitotic inhibitors, such as cytosine arabinoside (Ara-C), hydroxyurea (HU), with anti-Ig for the first 24 hr of incubation did not inhibit the production of IgG induced by anti-Ig and soluble factor. The addition of cytochalasin-B in the first stage of culture with anti-Ig did not inhibit, but rather enhanced, the formation of IgG. On the other hand, the presence of Ara-C, HU, or cytochalasin-B with soluble factor in the second stage of culture completely inhibited the production of IgG. Soluble factor preferentially induced the uptake of thymidine of anti-Ig-stimulated cells. Lipopolysaccharide (LPS) also induced the proliferation of rabbit lymphocytes, but did not induce the IgG production of anti-Ig-stimulated cells. However, LPS-stimulated cells formed a significant amounf of IgG when they were cultured with soluble factor. Anti-Ig-stimulated cells absorbed the enhancing activity of soluble factor, but non-stimulated cells did not absorb the activity. These results showed that cross-linkage of Ig receptors or LPS receptors with anti-Ig or LPS induced the formation of acceptor sites for soluble factor without the requirement of cell division and these activated cells started to proliferate and maturate to antibody-forming cells under the influence of soluble factor.


Triggering mechanism of B lymphocytes. II. Induction of ornithine decarboxylase in B cells by anti-immunoglobulin and enhancing soluble factor. In vitro stimulation of rabbit lymphocytes with anti-immunoglobulin antibody (anti-Ig) induced ornithine decarboxylase (ODC) (EC4.1.1.17) in the cells. The enzyme activity reached the maximum at 8 hr after stimulation of the cell with antibody and returned to the unstimulated level after 20 hr. The optimum concentration of antibody to produce the maximum activity of ODC was 25 mug/ml, which was in the same range that required for the maximum production of IgG and the highest rate of DNA synthesis. ODC was again induced with soluble factor (SF) in the cells that had been previously treated with anti-Ig antibody for 24 hr. The treatment of the cells with anti-thymocyte serum and complement did not affect the activity of the enzyme induced by anti-Ig or by SF, indicating that enzyme induction was an event occurring in B cells and not in T cells. The addition of mitotic inhibitors, such as hydroxy-urea (HU) or cytosine arabinoside (Ara-C), to the culture with anti-Ig did not inhibit the production of ODC. The presence of cytochalasin-B with anti-Ig did not inhibit, but rather enhanced, the synthesis of ODC. On the other hand, the presence of HU or cytochalasin-B with soluble factor completely suppressed the induction ODC. These results suggested that intracellular events that trigger the induction of ODC by the interaction of Ig receptor with anti-Ig may be different from the events that induce the same enzyme triggered by SF.


B lymphocyte differentiation from fetal liver stem cells in89 Sr-treated mice1. 89Sr treatment was used to determine if stem cells in young fetal liver could differentiate to mature B lymphocytes in adult animals that lacked functional bone marrow. Isotope injection alone resulted in a severe and lasting depletion of bone marrow cells and particularly lymphocytes, multipotential stem cells, and progenitors for granulocytes and macrophages. The latter two functional populations of cells expanded in number in the spleen but not the liver after their destruction in bone marrow. The total body content of stem cells did not, however, return to normal. Spleen B cells and lymphocytes lacking B or T cell markers were reduced by 89Sr treatment and possible reasons for this depletion are discussed. B lymphocytes were formed in lethally x-irradiated mice that had less than 1% of normal bone marrow function after reconstitution with fetal liver cells. These cells were immunologically mature by virtue of their ability to mount a primary immune response and function in a thymus-independent adoptive anti-hapten response.


Immune response to a syngeneic mammary adenocarcinoma. I. Comparison of kinetics of tumor cell growth and cytotoxic responses in syngeneic and allogeneic rats. The rat mammary adenocarcinoma 13762A is weakly immunogenic in syngeneic hosts. Transplantation immunity is not developed after injection of irradiated tumor cells. Two cell types were isolated from the tumor: one grew slowly forming solid implants; the second grew rapidly in ascites form, even in allogeneic hosts. Spleen cells cytotoxic for a tissue culture derivative of the tumor were produced in animals after injection of either cell type indicating a common antigen. The kinetics of tumor growth and production of cytotoxic spleen cells were compared in syngeneic and allogeneic animals. In vivo rejection of the tumor in allogeneic hosts did not correlate with the in vitro assay of spleen lymphocyte cytotoxicity. Futhermore, the cytotoxic response of spleen cells from syngeneic tumor-bearing hosts parallels that found in allogeneic hosts.


IgM antibody-dependent cell-mediated cytotoxicity in the Moloney sarcoma virus system: the involvement of T and B lymphocytes as effector cells. Antibody-dependent cell-mediated cytotoxicity in the Moloney sarcoma virus (MSV) system was analyzed with respect to the subpopulations of effector cells involved in tumor target cell destruction when IgM was used as the sensitizing antibody. With unfractionated sera from animals that had undergone regression primary MSV tumors it was found that macrophages did not contribute to the cytotoxicity induced by normal spleen cells that were syngeneic to the target cells. The IgM fraction of MSV regressor sera was found to induce cytotoxicity against the target cells by immunoadsorbent column-fractionated normal spleen cells, which were either depleted of T cells or B cells, according to the specificity of the columns. Immune IgM was also found to potentiate the activity of MSV regressor spleen cells that had been similarly fractionated. Furthermore, IgM antibody was found to induced cytotoxicity by normal spleen cells which had been depleted of either T or B cells by the appropriate antiserum (anti-T or anti-Ig) in the presence of complement and subsequent recovery of the viable cells by trysinization, filtration, and washing. However, spleen cells treated with both anti-T and anti-Ig sera simultaneously in the presence of complement and subsequet recovery of viable cells, were not induced to be cytotoxic against the IgM-coated tumor target cells. Further support oy T cells was provided by an experiment showing the induction with IgM of cytotoxicity against the target cells by normal thymocytes.


The humoral response of mouse spleen cells to two types of sheep erythrocytes. II. Evidence for gene expression in the B lymphocyte. There is multigene control of the response of mouse spleen cells to two types of sheep erytrocytes (H and L). Discriminator strains of mice make a much higher response to extra antigens found only on H SRBC than to the shared antigens found on both types of erythrocyte. Non-discriminator strains respond only to the shared antigens, making a responses as great as the discriminator response to the extra antigens. The cell type in which this genetic control is expressed was investigated by a number of methods for studying the role of the macrophage, T cell, and B cell in the production of anti-SRBC response. The results provide no evidence that the gene(s) operate at the macrophage, T cell, or any other accessory cell level. The data from B cell-limiting dilution experiments are compatible with genetic expression at the B cell level.


B cells in the appendix and other lymphoid organs of the rabbit: complement receptor lymphocytes. Complement receptor lymphocytes were demonstrated in rabbit lymphoid organs, the spleen containing the highest numbers (35%) and the thymus none. Extremely variable numbers (up to 50%) were detectablein the appendix. B and T cell depletion, rosette depletion, and histologic staining indicate that these CRL are B cells in the rabbit as in other species. The variable behavior of B cells in the appendix suggests a difference between this lymphoid organs population and those of other rabbit lymphoid organs.


Reappraisal of lymphocyte classification by means of surface morphology. Prior scanning electron microscope studies have indicated that T (thymus derived) lymphocytes appear "smooth" while human B (thymus independent) lymphocytes appear "hairy" with numerous microvilli covering their surfaces. A recent report (24) has raised some doubt as to the validity of classifying lymphocyte subpopulations on this purely morphologic basis. We have examined lymphocytes taken from congenitally athymic "nude" mice (lacking T cells) and compared them with those taken from normal littermates and CBA/J mice (both having T cell populations). We have determined that there are no distinguishable differences between the distribution of surface morphologies of lymphocytes from animals either with or without T and B cells. Our studies indicate that the classification of T and B cells, based on cell surface features, requires more than just finding out whether or not the cells are smooth or hairy.


Genetic complementation of histocompatibility-linked Ir genes in the rat. Different inbred rat strains that are congenic with respect to the major histocompatibility system (H-1) show either' a low, an intermediate, or a high immune response to the synthetic polypeptide (H, G)-A--L. Two out of four low responder H-1 haplotypes are found to complement intermediate responsiveness to high responsiveness. This effect is observed for the primary and secondary response and is detectable at the humoral and cellular level, i.e., antibody production and anamnestic response in vitro. Interallelic and intergenic complementation models are outlined. The ensuing implications for H-linked Ir gene control of immune responsiveness and for the structure of major histocompatibility systems are discussed.


Studies of thymic factors. I. Evaluation of the mouse rosette assay for thymic hormone. The restoration of sensitivity to azathioprine of athymic (nu/nu) mouse splenic rosette-forming cells (RFC) has been used by others to assay thymic hormone. These previous studies have led to the conclusion that a major factor in the development of autoimmunity in New Zealand (NZ) mice and patients with systemic lupus erythematosus (SLE) is the premature loss of thymic hormone. We have studied the mouse rosette assay and find that poly A-Poly U and single stranded DNA, in addition to thymosin, restored azathioprine sensitivity to nu/nu RFC. Thus, other substances may mimic thymic hormone, perhaps via the cyclic AMP system. Furthermore, antibodies to T cells were found to interfere with the assay. Sera from old NZ mice which contain naturally occurring thymocytotoxic antibodies (NTA) failed to restore azathioprine sensitivity. Small quantities of NTA prevented normal sera from functioning in the assay. We conclude that the role of thymic hormone(s) in the development of autoimmunity in NZ mice and patients with SLE must be completely reevaluated.


T-cell membrane characteristics of "mycosis cells" in the skin and lymph node. In some patients with mycosis fungoides atypical cells ("mycosis cells") are found in the blood. Recently the T-cell membrane characteristics of these atypical cells have been described. In this paper the results of a study of the atypical cells isolated from the lymph nodes and the skin lesions of three patients with mycosis fungoides are presented. Using electron microscopy, it could be demonstrated that the atypical cells formed rosettes with uncoated sheep red blood cells, but not with antibody-complement-coated sheep erythrocytes, indicating the T-cell membrane characteristics of the atypical cells.


Histiocytosis X of the larynx.

Histiocytosis X of the larynx. The present report is concerned with the observation of histiocytosis X in the larynx in a man aged 57 years. This localisation of the process is quite unusual and we have found only one similar case in the literature. The differential diagnosis and possible etiological factors are briefly discussed.


X-linked B-lymphocyte immune defect in CBA/N mice. II. Studies of the mechanisms underlying the immune defect. The mechanisms underlying the X-linked thymus-independent (B) lymphocyte functional defect in the CBA/N (CN) mice and their F1 progeny were studied. Immune defective mice were unable to respond to the T-independent antigen 2,4-dinitrophenyl-lysyl-derivative of Ficoll (DNP-lys-Ficoll) but were able to form antibody against the highly cross-reactive hapten (trinitrophenyl) when it was coupled to an erythrocyte carrier. Immune defective CN X DBA/2N (DN) F1 male mice, which do not normally respond to T-independent antigens, were able to respond to both polyribosinic-polyribocytidylic acid and DNP-lys-Ficoll after the administration of CN X DN F1 female spleen cells even if these cells had been depleted of T lymphocytes. In addition, it was shown that the inability of the CN mice and their F1 progeny to respond to T-independent antigens was not due to an intrinsic abnormality of their microenvironment or the suppressive actions of a T lymphocyte. Our data present evidence that the X-linked defect in the CN mice is due to an intrinsic defect in B-lymphocyte development.


The ontogeny of antigen-specific T cells. Although the fetal lamb is unable to form circulating antiovalbumin antibodies until about 120 days of gestation, ovalbumin-specific helper T cells can be stimulated to function at an earlier age. This would suggest that the critical event responsible for the precise sequential maturation of immunologic competence to different antigens at different developmental stages is not the first appearance of specific receptors on immunocytes. Alternative explanations of this phenomenon are discussed.


Localization of aggregated cell surface antigens of target cells bound to cytotoxic T lymphocytes. The redistribution of aggregated cell surface antigens of target cells bound to cytotoxic T lymphocytes was investigated. It was found that cap formation induced by antibody always occurred toward the site of binding. It is suggested that the polar localization of capped target cell surface determinants is a consequence of cytotoxic T-lymphocyte target cell interaction.


Kinetic study of T lymphocytes after sensitization against soluble antigen. III. Potentiation and suppression of the PHA response by antigen-activated lymphocytes of low density. Lymph node cells of ovalbumin-sensitized rats were separated on the basis of buoyant density into fractions reciprocally enriched in cells responsive to ovalbumin or phytohemagglutinin (PHA). Recombination of high density and low density fractions in varying proportions resulted in potentiation or suppression of the DNA synthetic response to PHA in culture. The response of cultures containing equal numbers of high and low density cells was markedly greater than the sum of the two populations stimulated separately. However, when decreasing numbers of low density cells were cultured with a constant number of high density cells, profound suppression was observed.


Clonal growth of hamster free alveolar cells in soft agar. Free alveolar cells obtained from healthy unstimulated hamsters were tested for their ability to form colonies in soft agar. Every bronchial washing so far tested contained colon-forming cells. The average plating efficiency was 8.1% (2.4-18.3%). Alveolar colony-forming cells were characterized by having a long initial lag period (4-8 days) and only mononuclear phagocytes were found in the colony. Medium conditioned by baby hamster kidney cells or other cells was required for the initiation and maintenance of their growth. Alveolar cells from normal mice and rats also formed colonies under appropriate culture conditions.


Coelomic fluid and blood serum proteins of the metamorphosing bullfrog tadpole. Bullfrog (Rana catesbeiana) tadpoles were divided into four groups containing Gosner ('60) stages as follows: I, 27-31; II, 32-36; III, 37-41; IV, 42-46. Extractable coelomic fluid volume from late metamorphic tadpoles (Group IV) was found to have a highly significant (p less than 0.001) negative correlation with stage; tadpole weight also had a negative correlation with stage (p less than 0.025). No significant correlation was found between coelomic fluid volume and tadpole weight. Group IV blood serum protein levels (x = 1.30 +/- SEM 0.10 G/100 ml serum) were shown to be significantly higher than corresponding coelomic fluid serum protein levels (x = 0.80 +/- 0.06 G/100 ml). Electrophoresis data demonstrated an increase in the albumin fraction of both blood serum and coelomic fluid during metamorphosis. I-125 labeled human serum albumin equilibrated with the blood serum at equal concentrations about ten hours after an intracoelomic injection. A coelomic fluid turnover time fo 26% per hour was calculated from the tracer data. We hypothesize that a slight sieving occurs at the coelomic fluid inflow site with none at the outflow site.


Fertilization of green frog (Rana clamitans) eggs in their native jelly by bullfrog (Rana catesbeiana) sperm. Fertilization of R. clamitans eggs in their native jelly occurred when inseminated with high concentrations of R. catesbeiana sperm. Contrary to previous evidence, the block to cross-fertilization of the R. clamitans egg is not complete. This raises the possibility that R. clamitans x R. catesbeiana hybrids may form in nature.


Structural dynamics of frog muscle during isometric contraction. Intensity fluctuation autocorrelation functions of laser light scattered by actively contracting muscle were measured at points in the scattered field. They were reproducible and showed characteristics which depended on the physiological state of the muscle and the parameters of the scattering geometry. The autocorrelation functions had large amplitudes and decay rates that varied significantly with the phase of the contraction-relaxation cycle. The dependence of the autocorrelation function on scattering geometry indicated many elements with diameters on the order of 0.5 mum (presumed to be myofibrillar sarcomeres or their A bands or I bands) undergo independent random changes in their axial positions and their internal distribution of optical polarizability during the plateau of an isometric tetanus. The experimental results are interpreted in terms of a model in which most of the scattering elements in isometrically contracting muscle have random fluctuating axial velocities of average magnitude 20 nm/ms that persist for a few milliseconds at least. In addition to these axial motions there are local fluctuations in polarizability. Similar intensity fluctuation autocorrelation functions were observed throughout the active state on two muscle preparations, whole sartorius muscle and small bundles of single fibers (three to eight) of semitendinosus muscle. These results imply that the tension developed during an isometric tetanus contains a fluctuating component as well as a constant component.


Actions of external hypertonic urea, ADH, and theophylline on transcellular and extracellular solute permeabilities in frog skin. Increases in transepithelial solute permeability were elicited in the frog skin with external hypertonic urea, theophylline, and vasopressin (ADH). In external hypertonic urea, which is known to increase the permeability of the extracellular (paracellular) pathway, the unidirectional transepithelial fluxes of Na (passive), K, Cl, and urea increased substantially while preserving a linear relationship to each other. The same linear relationship was also observed for the passive Na and urea fluxes in regular Ringer and under stimulation with ADH or 10 mM theophylline, indicating that their permeation pathway was extracellular. A linear relationship between Cl and urea fluxes could be demonstrated if the skins were separated according to their open circuit potentials; parallel lines were obtained with increasing intercepts on the Cl axis as the open circuit potential decreased. The slopes of the Cl vs. urea lines were not different from that obtained in external hypertonic urea, indicating that this relationship described the extracellular movement of Cl. The intercept on the ordinate was interpreted as the contribution from the transcellular Cl movement. In the presence of 0.5 mM theophylline or 10 mU/ml of ADH, mainly the transcellular movement of Cl increased, whereas 10 mM theophylline caused increases in both transcellular and extracellular Cl fluxes. These and other data were interpreted in terms of a possible intracellular control of the theophylline-induced increase in extracellular fluxes. The changes in passive solute permeability were shown to be independent of active transport. The responses of the active transport system, the transcellular and paracellular pathways to theophylline and ADH could be explained in terms of the different resulting concentrations of cyclic 3'-5'-AMP produced by each of these substances in the tissue.


Resolution of the potassium ion pump in muscle fibers using barium ions. When frog sartorius muscles recover from Na enrichment in the presence of external K, net K entry into the fibers occurs by both passive movement and active inward transport via a K pump. Under normal conditions, it has not been possible to experimentally distinguish these processes. Fractionation into the flux components must be accomplished from inferences concerning the K conductance or permeability during a period of rapid Na extrusion. The best estimates indicate that 60-80% of the K entry occurs via the K pump. In the presence of Ba ions, the membrane permeability to K is very much reduced. Under these conditions, Na-enriched muscles underwent a normal recovery in the presence of external K, and the amount of inward K movement due to the K pump rose to over 90% of the total K entry. The characteristics of the K pump studied by this means were: (a) essentially complete inhibition by 10(-4) M ouabain, (b) inhibition by [Na]omicron, (c) activation by [K]omicron according to a rectangular hyperbola in the absence of [Na]omicron, (d) linear activation by [Na]iota over a wide range in concentration, (e) zero or undetectably low pumping rate as [Na]iota leads to 0, (f) the number of Na ions actively transported per K ion actively transported is 1.4-1.7 normally and 1.1 in the presence of Ba.


Variation of muscle stiffness with force at increasing speeds of shortening. Single frog skeletal muscle fibers were attached to a servo motor and force transducer by knotting the tendons to pieces of wire at the fiver insertions. Small amplitude, high frequency sinusoidal length changes were then applied during tetani while fibers contracted both isometrically and isotonically at various constant velocities. The amlitude of the resulting force oscillation provides a relative measure of muscle stiffness. It is shown from an analysis of the transient force responses observed after sudden changes in muscle length applied both at full and reduced overlap and during the rising phase of short tetani that these responses can be explained on the basis of varying numbers of cross bridges attached at the time of the length step. Therefore, the stiffness measured by the high frequency legth oscillation method is taken to be directly proportional to the number of cross bridges attached to thin filament sittes. It is found that muscle stiffness measured in this way falls with increasing shortening velocity, but not as rapidly as the force. The results suggest that at the maximum velocity of shortening, when the external force is zero, muscle stiffness is still substantial. The findings are interpreted in terms of a specific model for muscle contraction in which the maximum velocity of shortening under zero external load arises when a force balance is attained between attached cross bridges somr interpretations of these results are also discussed.


Attempts to measure endogenous levels of dopa and 5-hydroxytryptophan in rat brain. We have used previously published analytical methods of high sensitivity, developed in our laboratory, in an attempt to detect endogenous dopa and 5-hydroxytryptophan (5-HTP) in normal rat brain. In the brains of rats killed by decapitation we were unable to detect any dopa and found only a few ng/g of 5-HTP. Higher levels were found in the brains of rats killed by exposing their heads to microwaves. The levels of dopa found under these conditions varied considerably between different rat strains, and also between different experiments on the same strain, ranging between 2.5 and 13.8 ng/g. True resting levels may be lower than those observed because the procedure necessitated a short-lasting immobilization stress which increased the dopa (but not 5-HTP) levels. The 5-HTP levels were also variable and ranged between 5 and 25 ng/g. The highest levels of dopa and 5-HTP were found in the Japanese Wistar strain with spontaneous hypertension. Some analytical pitfalls are pointed out.


[Pineal organ and parietal eye of Lacerta vivipara: a qualitative and quantitative microscopic study of the uptake of 3H-5-hydroxytryptophan during ontogenesis]. After comparison of results obtained by different modalities of 3h-5HTP injections (involving intraallantodic, intraventricular and intraperitoneal injections), the intraperitoneal one was retaineed to study experimental cytochemcal biosynthesis of indoleamines, in the epiphysis of Lacerta vivpara J., during ontogenesis. From stage 36 of embryogenesis, the selective radioautographic labelling, significant of atritiated indoleamines storage, was located in the distal part of the epiphysis. From stage 36 to 40 (just before hatching) and 3 days after hatching, microscopic and microphotometric studies point out, in the epiphysis epithelium, a highly significant increase in tritiated infoleamines rate. In contrast, the weak nonspecific reactions, in the neighbouring brain structures (dorsal telencephalon, tectum opticum and paraphysis), stay constant. The metabolism and storage sites of indoleamines belong to sensory line cells. During ontogenesis, these sites appear successively in distal, middle and proximal epiphysis and from the basal to the upper part of the sensory line cells. The embryonic parietal eye, in spite of structural analogies, does not take up the radioactive compound.


The life cycle of Haematoloechus coloradensis Cort 1915 (Digenea: Plagiorchiidae), with emphasis on host susceptibility to infection. Mother and daughter sporocysts and xiphidiocercariae of Haematoloechus coloradensis develop in the snail, Physa virgata. Cercariae penerate and encyst in dragonfly nymphs, Tramea sp., Libellula sp., Anax sp., and in damselfly nymphs, Enallagma spp. Upon ingestion by the vertebrate host, metacercariae excyst and migrate into the lungs where they mature. Haematoloechus coloradensis is specific for its definitive host, R. pipiens, but shows some development in Ambystoma tigrinum. Although H. coloradensis is specific for its snail host, all odonate species tested served equally well as the second intermediate host.


The effect of temperature on desensitization kinetics at the post-synaptic membrane of the frog muscle fibre. 1. The time course of acetylcholine (ACh) potentials during development of desensitization was prolonged when a double-barrel ACh pipette was used for evoking desensitization. When a single-barrel ACh pipette was used, no change in the time course of ACh potential was to be seen during desensitization. 2. The recovery after desensitization did not depend on the rate of onset or on its level when a single ACh pipette was used. The half-time of recovery had a constant value of about 5.8 sec in the presence of chlorpromazine or SKF-525 A in a muscle bath at 22 degrees C. 3. Unlike the rate of onset, recovery from desensitization does not depend on the membrane potential. 4. The rate of onset of desensitization, i.e. time taken for reduction of ACh potentials to half-way between the initial amplitude and final steady value, decreased when temperature of the muscle bath was lowered. 5. Q10 of desensitization onset was found to be 1.5 for a change of temperature from 32 to 22 degrees C, 1.9 from 22 to 12 degrees C, 2.6 from 12 to 5 degrees C and 3.3 from 12 to 2 degrees C. 6. A similar temperature effect was observed in the case of desensitization recovery, the Q10 being 1.2 for temperature changes from 32 to 22 degrees C, 1.3 from 22 to 12 degrees C and 2.36 from 12 to 2 degrees C. 7. Intracellular application of quaternary methiodide of SKF-525 A or chloropromazine caused more rapid desensitization by ACh. The rate of desensitization onset depends on the ACh dose and on the frequency of application. The rate of recovery, however, has a constant value with a half-time of 5.5-5.7 sec at 22 degrees C. 8. Both the rate of onset and the rate of recovery changed with temperature in the case of intracellular potentiation of desensitization, in a similar manner to that observed after extracellular application of these drugs. 9. The onset of desensitization can thus be influenced by different substances as well as by changes in temperature. Recovery apparently has a different mechanism from the onset, because its time course can be altered only by changes in temperature of the muscle.