Rabbit lymphocyte populations responding to haptenic and carrier determinants for DNA synthesis. Mesenteric lymph node cells from rabbits primed with dinitrophenyl derivative of either Ascaris extract (DNP-Asc) or ragweed fraction D (DNP-Rag) were stimulated by the priming antigen, free homologous carrier, or with the hapten conjugated with rabbit serum albumin (DNP-RSA), and increase in DNA synthesis was observed by 3H-thymidine incorporation. The results showed that both free carrier and DNP-RSA stimulated DNA synthesis. The response of the primed lymph node cells to DNP-homologous conjugate was slightly higher than that to free carrier, but the optimal concentration of both antigens for maximal thymidine incorporation was 10 to 100 mug/ml. This concentration was about 100 times higher than the optimal concentration of the same antigens for maximal antibody response in vitro. The DNA synthetic response to DNP-RSA was much less than that obtained by free carrier, and the optimal concentration of DNP-RSA for the response was comparable to the concentration of DNP-homologous carrier conjugate to induce maximum anti-DNP antibody response. The relative importance of immunoglobulin bearing (B) cells and T cells in the DNA synthetic responses was assessed by fractionating lymph node cells with antigen-coated or anti-immunoglobulin coated  cellular immunosorbent. The results indicated that hapten-specific, immunoglobulin-bearing cells (B cells) are responsible for the DNA-synthetic response to DNP-RSA, whereas B cells play a minimal role in the response to free carrier.


Competition between concanavalin A-induced stimulatory and inhibitory effects in the in vitro immune response to antigen. The humoral response of nude spleen cells (b cells) to sheep erythrocytes was measured in the presence of varying numbers of concanavalin A (ConA)-acvated stimulatory spleen T cells (helper) and Con A-activated inhibitory spleen T cells (suppressor) from BDF1 mice. It was found that suppressive effects could be reversed by the presence of additional numbers of stimulatory cells. These results seem incompatible with the hypothesis that suppression is mediated by supraoptimal numbers of stimulatory cells and provides additional evidence that separate populations of T cells mediate stimulation and suppression.


Corneal thickness measured by interferometry. An optical method for measuring the thickness of transparent structures has been developed, and has been used to measure, in vivo, the thickness of the human cornea. The thickness is measured by placing the anterior surface of the cornea at the focus of a beam of coherent laser light and then measuring the spacing between the interference fringes generated by the reflected light. The thickness is then calculated from the fringe spacing. The method has been used to measure corneal thickness in frog and human corneas. These measurements have been correlated with histologic and pachometer measurements of corneal thickness. A significant capability of this technique is to measure the thickness of optically opaque corneas.


Runt syndrome induced in the rat by polyinosinic-polycytidylic acid. Polyinosinic with polycytidylic acid (poly l with poly C), a double-stranded synthetic RNA, produced in newborn rats a runt syndrome characterized by mortality and retarded growth rates of the total body, thymus, and kidneys. In contrast, it induced a hyperplasia in the epidermis and in the spleen. Within 10 days of treatment, the epidermis became 2 or 3 times thicker and the spleen mass was increased by 50%. The epidermal hyperplasia involved all layers, but hair follicles were excluded. Splenic hyperplasia did not result from accelerated erythropoiesis. Double-stranded RNA was required; single-stranded homopolymers were ineffective. Theophylline, a phosphodiesterase inhibitor, did not potentiate the effects. The uptake of iododeoxyuridine-125 was not enhanced in the hyperplastic epidermis or spleen. Thus we concluded that poly l with poly C can retard the growth of some organs in newborn rats, but that it causes epidermis and spleen to accumulate cells. The cytokinetic mechanisms involved in these contrasting effects were not clear.


Tumor induction by Moloney sarcoma virus in athymic nude mice. Athymic nude (nu/nu) mice, normal NMRI (plus/plus) mice, and nu/plus heterozygotes, inoculated at birth with the Moloney strain of murine sarcoma virus (M-MSV), did not differ either in tumor growth rate or incidence or in mortality due to tumor development; neither did nude (nu/nu) and normal NMRI mice inoculated with M-MSV at 30 days of age differ in the initial tumor growth rate. However, tumors induced in adult nude mice grew progressively and eventually killed the hosts, whereas tumors induced in adult normal NMRI mice completely regressed. These results indicated that thymus-dependent immunity, though not active in M-MSV tumor induction, determined whether the tumor, once formed, would regress or kill the host.


Laser-induced hemostasis in the canine stomach. Use of a flexible fiberoptic delivery system. A flexible fiberoptic system has been used to deliver visible laser light into the upper gastrointestinal tract to arrest bleeding from various induced lesions. This procedure has achieved hemostasis at bleeding sites in the gastrointestinal tract of animals and suggests that bleeding lesions in the heman may be treated similarly without resort to surgical intervention or intensive medical management.


Antitrypsin phenotypes in St. Louis.

Antitrypsin phenotypes in St. Louis. Antitrypsin phenotypes were determined from 2,285 donors at the Barnes Hospital Blood Bank in St. Louis . Results resembled those obtained from Central Europeans: there were fewer proteinase inhibitor (Pi) Z alleles in St. Louis than in Scandinavian test groups. Interestingly, major antifrypsin variants occurred only 40% as frequently in black as in the balance of the study group.


Open-heart surgery in von Willebrand's disease. The case is presented of a hemophilia carrier, also affected by von Willebrand's disease, who underwent aortic valve replacement. The clinical and laboratory findings of von Willebrand's disease (prolonged bleeding time, low factor VIII, and abnormal platelet activity) are discussed, and a protocol for management of patients with low factor VIII levels (such as hemophilia carriers and subjects with hemophilia A or von Willebrand's disease) undergoing open-heart surgery is proposed. Our case proves that corrective cardiac surgery in similar circumstances is feasible so long as adequate levels of factor VIII are maintained.


[Effect of acceleration on the sugar content of rabbit cerebrospinal fluid]. The sugar content in the liquor of rabbits exposed to accelerations ranging from several minutes to several hours was studied. The animals were accelerated at 3 g in a centrifuge with a radius of 55 cm and a velocity of 70 rpm, in the Khilov's swing and in a rotating device simulating angular accelerations. The liquor was sampled by the suboccipital puncture and the sugar content was measured according to Hagedorn-Jensen. A statistically significant increase in the sugar content was noted after every exposure to accelerations of alternating signs than with accelerations of a constant value. The experiments on labyrinth-ectomized animals showed a certain effect of the vestibular analyzer on the carbohydrate metabolism. However, when the equilibrium function reached compensation, the sugar content in the liquor returned to the pretest level. Therefore, this reaction involves not only the vestibular system but also the optic, proprio- and interoceptive afferent systems. A comparison of the intracysternal and intramuscular injections of epinephrine points to the central (in contrast to the hematogenic) nature of an increase in the sugar content during the first minutes of acceleration. The electric stimulation of the anterior and posterior hypothalamus by means of implanted electrodes produced the same elevation of the liquor sugar as accelerations. It can be postulated that afferent impulses from the analyzer systems reach the hypothalamic centers, thus exciting the autonomic system and, first of all, the sympathic system.


Results of direct coronary artery surgery in women. In the past 4 years, among 260 patients receiving coronary bypass grafts for coronary artery disease and stable angina pectoris, there were 34 women (13 per cent). The operative mortality rate for women was 8 per cent (3 of 34), and one late death due to myocardial infarction occurred in spite of a patent coronary bypass graft. The intraoperative infarction rate was 20 per cent (6 per cent in men). Although preoperative cardiac pump and muscle function parameters were better in women than in men (p less than 0.05), postoperatively only 30 per cent of women showed improvement in function as compared with 50 per cent of men. At 6 to 46 months' follow-up, 84 per cent of women were free of angina in contrast with 94 per cent of men. The early (4 month) graft patency rate was 50 per cent (14 of 27 grafts), as opposed to 80 per cent (20 of 25 grafts) in men. These results indicate that, although coronary artery disease shows anatomic similarity in women and men, the result of coronary revascularization in women is inferior to that in the male population.


Survival and function after left ventricular aneurysmectomy. Operative and late postoperative results in 55 patients undergoing resection of a discrete, saccular type left ventricular aneurysm are described. All patients had occlusive lesions of either the left anterior descending or left main coronary artery, and 72 per cent had significant multivessel disease. Preoperative factors that correlated significantly with decreased long-term survival were age greater than 60 years, severe congestive heart failure, and subnormal cardiac output at rest. The primary determinant of long-term postoperative survival appears to be the functional state of residual myocardium after aneurysmectomy. The survival rate for patients with multivessel disease was not less than that of patients with isolated lesions of the left anterior descending coronary artery. The impact of simultaneous aorto-coronary bypass grafting on postoperative results in patients with multivessel coronary disease is discussed. An advantage to concomitant revascularization with aneurysmectomy cannot be statistically corroborated in this series. However, improvement in results since the routine application of bypass grafting in appropriate cases, the uniform operative survival of bypassed patients with triple vessel disease, and the relatively high frequency of myocardial infarction as a cause of late postoperative death suggest that bypass grafting in patients with multivessel disease should be combined with aneurysmectomy to maximally improve long-term prognosis.


Surgical treatment for stable angina pectoris. Prospective randomized study. To evaluate the role of coronary bypass treatment of stable angina, 72 patients were randomly divided into surgical and medical groups. They were followed with treadmill tests and repeat catheterization. Anginal symptoms decreased or disappeared in 89 per cent of those operated on and 65 per cent of those not operated on, but more of the former than of the latter became asymptomatic: 69 vs. 11 per cent respectively (p smaller than 0.01). Exercise tolerance time improved significantly (p smaller than 0.001) in both groups, but more in the group operated on, plus 78 per cent vs. plus 48 per cent, p smaller than 0.05. During a 28-month follow-up period, fewer cardiac complications occurred in the patients operated on--14 vs. 34 (p smaller than 0.02). Mortality was 9 per cent in the patients operated on and 14 per cent in those not operated on; this difference is not significant. The results indicate that improvement, though demonstrable in both groups, is greater with surgery, and quality of life during the first 28 months is better in patients who are operated on.


Clinical importance of lymphoblasts with T markers in childhood acute leukemia. Of 48 children with acute lymphocytic leukemia 11 had blast cells with receptors for sheep erythrocytes in their initial bone-marrow aspirates and 37 did not. A comparison of selected clinical features indicated striking differences between the two groups. Leukemia with the receptors was associated with a high proportion of older children, predominantly boys, a thymic mass, and a high white-cell count at diagnosis. In contrast, the 37 children with leukemia without the receptors were generally less than five years of age, with a nearly equal distribution of boys and girls; all but one had normal chest roentgenograms, and only one had a white-cell count greater than 100,000. Thus, the presence or absence of lymphoblasts with sheep erythrocyte receptors--a T-cell marker--distinguishes two forms of childhood acute lymphocytic leukemia, each with a distinct distribution of age and sex as well as other characteristic clinical features.


Abnormal spontaneous rosette formation and rosette inhibition in lung carcinoma. Fifty-five children with CID and known ADA status were studies at a workshop held in Albany, New York. Erythrocyte ADA determinations were performed in 22 of the 55 patients, 13 of whom were ADA negative. The ADA defect appears to be transmitted as an autosomal recessive trait. Some patients with CID and ADA deficiency have characteristic radiologic abnormalities of the skeleton, which are not found in other illnesses. The thymus glands of all patients with CID and ADA deficiency who could be examined have evidence of thymic involution manifested by presence of Hassall's corpuscles and differentiated germinal epithelium; this is in contrast to "classic" thymus findings in CID with normal ADA. Adenosine deaminase probably plays an important, although as yet undefined, role in lymphocyte development and/or function. The deficiency of ADA in CID is the first enzyme defect observed in a deficiency disease of specific immunity.


Therapy of intention myoclonus with L-5-hydroxytryptophan and a peripheral decarboxylase inhibitor, MK 486. Three patients with postanoxic intention myoclonus, two patients with intention tremor, and one patient with cerebral palsy were administered L-5-hydroxytryptophan (L-5HTP), the precursor of serotonin, in combination with MK 486, a peripheral amino acid decarboxylase inhibitor. L-5HTP combined with MK 486 were potent long-term therapeutic agents for postanoxic intention myoclonus, but had no effect on intention tremor or cerebral palsy. These drugs were well-tolerated by the patients, and more effective than any other known therapy for intention myoclonus. Cerebrospinal fluid concentration of 5-hydroxyindoleacetic acid, the main catabolite of serotonin, appeared low in two patients with intention myoclonus and increased markedly during drug therapy. Postanoxic intention myoclonus may be causally related to a deficiency of brain serotonin.


Vestibulo-ocular function in patients with cerebellar atrophy. Eye movement abnormalities were quantiatively assessed in four patients with clinically pure cerebellar atrophy (group A), six patients with brainstem plus cerebellar atrophy (group B), and five patients with Friedreich's ataxia (group C). Twelve patients had one or more types of spontaneous nystagmus; eight gaze nystagmus, three rebound nystagmus, wo positional nystagmus, and one vestibular nystagmus. Catoric-induced and rotatory-induced nystagmus was hyperactive in group A and diminished in group C. Group B had mixed responses. All patients demonstrated significant fixation instability and impaired smooth pursuit. There was dysmetria of voluntary saccades, with flutter and "rebound" saccades. Nine of 15 patients had significant slowing of induced saccades, including two patients in group A. It is concluded that quantitative vestibulocular tests can be useful in classifying the cerebellar atrophy syndromes.


[Immunological aspects of Hodgkin's disease]. Immunological test alterations in Hodgkin's disease are described. Cutaneous reactivity to various antigens (tuberculin, oidiomycin, trichophytin, parotitis virus, candida, etc.), cutaneous reactivity to sensitizing chemical agents (DNCB, DNFB), lymphocyte blastization in vitro with PHA and various antigens, the reaction of homologous lymphocytes transferred to the skin, and the stimulation by the Hodgkin lymphocytes of homologous lymphocytes in unidirectional mixed culture, are all impaired. Antibody immunity would appear to be normal. The impairment of the T system and normality of the B system would explain these patients' much greater susceptibility to viral than to bacterial diseases. On the basis of the most recent research, it is hypothesized that underlying these phenomena is a deficiency in the H-LA system which makes it unable to recognize certain neoplastic antigens (Reed-Strenberg reticular cells, for example); stimulation of the B system by the formation of anti-T antibodies and subsequent depletion of this system.


Differences in action potentials and accommodation of sensory and motor myelinated nerve fibres as computed on the basis of voltage clamp data. 1. Voltage clamp experiments were performed on sensory and motor nerve fibres of the frog using a digital computer for automatic experiment control and data recording. 2. Rates of rise and maximum amplitudes of potassium currents were determined in both sensory and motor fibres, so that comparative values of n infinity and tau-n could be obtained. 3. The results indicate that the n infinity-V curve for sensory fibres is displaced from the curve for motor fibres in a depolarising direction. The potassium kinetics are similar in both for voltage steps up to about - 20 m V, beyond which tau-n becomes progressively smaller for sensory than for motor fibres. 4. These comparative values of n affinity and tau-n have been used to calculate alpha-n and beta-n values for a model "motor" action potential by considering the Frankenhaeuser-Huxley computed action potential to be "sensory". This modification of the potassium system, together with some alteration to the sodium inactivation system produces a satisfactory model "motor" action potential. 5. The model sensory and motor action potentials behave quite similarly to their experimentally recorded counterparts with respect to action potential shape and relative duration, repetitive firing, accomodation and the simulated action of T.E.A.


Early receptor potential from the pineal photoreceptor. Rapid photoresponses were obtained from the pineal organ of Rana catesbeiana and Rana esculenta comparable to the early receptor potential (ERP) of the retina of their lateral eyes. Light source was a xenon filled discharge tube with a maximum energy input of 365 J. The flash duration was 600 mu sec measured at 1/3 peak amplitude. The response pattern of the ERP of the exposed Epiphysis cerbri (intracranial portion of the frog's pineal organ) resembled the ERP of the isolated retina and the R1 deflection of the ERP of the eye-cup. It was greatly diminished during consecutive flashes and was absent during light adaptation. It was confined to the Epiphysis cerbri and was similar in time course and shap to, though smaller as, the ERP of the isolated retina. The photoresistant component of the ERP in the eye-cup was lacking in the Epiphysis cerbri and in the isolated retina. The action spectrum of the ERP of the Epiphysis cerbri closely resembled the photopigment absorption of rhodopsin indicating that the structural cones of the pineal organ contain a photopigment different from the retinal cones of the frog's lateral eye.


[99mTc-polyphosphate scintigraphy of joints, xeroradiography and radiography of hands]. Joint imaging with 99m-Tc-Polyphosphate was carried out in 21 patients having inflammatory or degenerative diseases of the joints of the hands. A comparison was made between 99m-TcPP, xeroradiography and radiography. In a total of 630 different joints 237 positive scans, 132 positive roentgenograms and 183 positive xeroradiograms were obtained. In the early stage of inflammatory joint disease only the scintigraphic results are positive, in the later stage 99mTcPP-imaging was less sensitive than xeroradiography and radiography. The reason for the increased concentration at the site of acute joint inflammation is probably the increased uptake in the soft tissues of the joints. The best time for scanning after the application of 99m-TcPP seemed to be at 3 hours.


Mitogen-induced blastogenic responses of lymphocytes from marihuana smokers. Blastogenic responses in vitro to phytohemagglutinin and pokeweed mitogen were examined in microcultures of peripheral blood lymphocytes from a group of 12 healthy, long-term marihuana smokers and a group of matched control subjects. With either mitogen, no significant difference in cellular incorporation of (3H)thymidine was noted between the groups. These results were interpreted to indicate that the functional status of blood lymphocytes was not altered by long-term smoking of marihuana.


Complement-fixing and lymphocytotoxic antibodies in serum of pregnant women at delivery II. Clinical observations I. Sera from 2,008 pregnant women were examined for the presence of complement-fixing platelet antibodies (CFPAb) and lymphocytotoxic antibodies (LCTAb). Women with antibodies in their serum were compared with women without antibodies. No clinical significance of CFPAb or LCTAb was found as regards pre-eclampsia, arterial hypertension, albuminuria, oedema, glucosuria, premature delivery, spontaneous onset of labor and abortion.


Coronary artery surgery for recurrent ventricular arrhythmias in patients with variant angina. Two patients with Prinzmetal's variant angina had recurrent episodes of resting chest pain, ST segment elevation, QRS widening, ventricular tachycardia, and ventricular fibrillation. These eqisodes were unresponsive to medical therapy including lidocaine, procaine amide, and quinidine sulfate. Coronary arteriography revealed severe obstructive coronary artery disease, involving more than one coronary artery, in both patients. Aorticocoronary saphenous vein grafts were utilized to bypass significant disease in each patient. In one patient blood flow through the grafts was measured at 90 and 65 ml per minute, respectively, at operation and patent grafts were demonstrated six months postoperatively. Neither patient has had recurrence of chest pain or evidence of ventricular tachycardia at one year or 2 1/2 years postoperatively. Postoperative resting and maximal exercise ECG's are normal. Coronary artery surgery may be an effective method of therapy for ischemic ventricular tachycardia when medical therapy fails.


Evaluation of ristocetin-Willebrand factor assay and ristocetin-induced platelet aggregation. Normal subjects, patients with various bleeding disorders, and patients with von Willebrand's disease were studied. All patients with von Willebrand's disease had decreased levels of ristocetin-Willebrand factor (range, 0 to 41%) as compared with all other subjects (range, 79 to 202%). Ristocetin-induced platelet aggregation of platelet-rich plasma was abnormal in all patients with von Willebrand's disease tested, and it was possible to correct this abnormal response by addition of normal platelet-poor plasma. Abnormal ristocetin-induced platelet aggregation was seen in patients with intrinsic platelet disorders or, on some occasions, in normal patients who had ingested aspirin. Ristocetin-induced platelet aggregation is not diagnostic, but it may be useful as a simple screening test for patients with possible von Willebrand's disease. In conjunction with other tests, the assay for ristocetin-Willebrand factor will be useful in diagnosis and evaluation of these patients.


Serum trypsin inhibitory capacity and Pi phenotypes. I. Methods and control values. Serum trypsin inhibitory capacity determinations are of considerable value in detecting genetically determined types of obstructive pulmonary disease and hepatic disease. These determinations must frequently be followed by determination of protease inhibitor (Pi) phenotype in order to confirm the diagnosis. Piphenotyping has been a specialized and time-consuming procedure, and suggested improvements in the methodology and technics may make it more generally applicable as a clinical laboratory procedure. The prevalence of phenotypes other than MM in a group of 700 control sera from blood donors is reported as a baseline to evaluate typically American populations of mixed ethnic and racial characteristics. There are suggestive differences in prevalences of S and Z genes relating to ethnic stock and racial groups. It is important when comparing the prevalences of S and Z genes in diseasedpopulations to use control groups of similar ethnic and racial compositions. Pi phenotyping is a necessary laboratory procedure in the diagnosis of certain forms of genetically determined chronic obstructive pulmonary disease and hepatic disease. The distributions of all serum protease inhibitory capacity values and those for S and Z Pi phenotypes are shown.


Enhanced rate of progression of coronary artery disease following aortocoronary saphenous vein bypass surgery. To evaluate the fate of the boronary arteries after aortocoronary bypass, 40 patients underwent serial selective coronary angiographic studies 1 year apart, and the frequency of progression of coronary artery disease was estimated. Thirty-two had saphenous vein bypass surgery after the first procedure, six had Vineberg operations, and two had no interim operation. In each patient, the right, left, anterior descending and circumflex coronary arteries (including their branches) were separately evaluated. Progressive narrowing was evident in 31 of 50 (62 per cent) bypassed vessels and in only 11 to 113 (9.7 per cent) nonbypassed arteries (p less than 0.001). Coronary arteries with moderate to severe obstruction initially (50 to 99 per cent occluded) manifested progressive diseasee more frequently (33 of 70 arteries) than did arteries that were normal or mildly narrowed initially (4 of 71) (p less than 0.001. Considering only those vessels with 50 to 99 per cent obstruction initially, 27 of 35 (77 per cent) of the bypassed arteries and only 6 of 35 (17 per cent) of the nonbypassed arteries showed progression (p less than 0.001). We conclude that moderately or severely narrowed coronary arteries are more likely to show progressive narrowing than normal or mildly obstructed ones and that progression of coronary disease is greater in bypassed vessels than in nonbypassed vessels. In view of potential graft closure, the implications of these findings must be considered in selecting patients for aortocoronary bypass.


Coronary bypass surgery for unstable angina pectoris. Clinical follow-up and results of postoperative treadmill electrocardiograms. The first 81 patients who underwent coronary artery bypass surgery at Stanford University Hospital for unstable angina pectoris have been followed up for an average of 18 months. The over-all surgical mortality was 8.6 per cent (seven patients). There have been no operative deaths in last 32 patients, which may be due to over 75 per cent of these patients being stabilized on intensive medical therapy from 24 to 72 hours before study or surgery. There was a 16 per cent (13 patients) perioperative and 15 per cent (12 patients) late incidence of myocardial infarction. Of 74 patients who survived the initial operation 2 died 2 and 3 months postoperatively. Good or complete relief from pain was obtained in 94 per cent (70 patients) of the survivors. Of 57 longterm survivors tested, 49 per cent (28 patients) had a definite ischemic response to treadmill exercise testing. This may reflect the severe nature of the occlusive coronary disease or mechanisms other than increased coronary flow being responsible for the relief of pain. Although coronary bypass surgery appears to be effective in relieving the pain of patients with unstable angina pectoris, the 18 month average follow-up indicates that the incidence of myocardial infarction in surgically treated patients is comparable to that in medically treated patients.


Immunologic and morphologic studies of T cell lymphoma. Thymic-independent (B) lymphocytes, thymic-independent (T) lymphocytes and histiocytes may be distinguished by the presence of certain surface markers. In addition B and T lymphocytes have been reported to show distinctive surface architecture by scanning electron microscopy. Neoplastic cells from a lymph node and cerebrospinal fluid of a patient with a diffuse malignant lymphoma of the poorly differentiated lymphocytic type were examined in frozen sections and cell suspensions for the presence of surface immunoglobulin and the antigen-antibody-complement (IgMEAC) receptor of B lymphocytes, the presence of the cytophilic antibody (IgGEA) receptor of histiocytes and the ability to form nonimmune rosettes with sheep red blood cells (E) characteristic of T lymphocytes. Cells from the lymph node were also studied by scanning electron microscopy. The majority of neoplastic cells from the lymph node and cerebrospinal fluid formed rosettes with E, but lacked surface immunoglobulin and failed to bind IgMEAC or IgGEA. By scanning electron microscopy the neoplastic cells, although larger in diameter, showed surface architecture similar to normal lymphocytes with a varying number of surface microvilli. These studies suggest that the malignant cells of this lymphoma are of thymic type.


Hemolytic anemia associated with multiple autoantibodies and low serum complement. A 37 year old woman with extravascular hemolytic anemia had a positive Monospot test associated with positive antiglobulin and anticomplement Coombs' tests, cold agglutinins and warm autoantibodies. IgG-kappa (k) antibodies, which reacted with all panel red cells at 37 degrees C, were eluted from her circulating red cells. However, neither immunoglobulins nor C3 was detected after her serum was adsorbed with heterologous red cell stroma at 37 degrees C and eluted at the same temperature in glycine buffer. In contrast, IgM-kappa and IgM-lambda (lambda), IgG-3-kappa, IgG4-lambda, IgA-lambda and C3 were eluted at 37 degrees C from heterologous red cell stroma after adsorption with her serum at 0 degrees C. Thus, antibodies of several types, which were present in the patient's serum, reacted optimally with red cell antigens at low temperature. Cold-reactive IgG3-kappa antibodies, which also capable of interacting with red cells at 37 degrees C, probably accounted for the IgG-kappa antibodies eluted from the patient's circulating red cells. The patient's serum C4 titers were decreased, with low normal to moderately depressed C3 and low normal C5, indicating that the anti-red cell IgM and/or IgG3-kappa antibodies probably fixed complement. A localized cold stress test resulted in a transient increase in plasma hemoglobin and a decrease in serum C3 titer. These findings, and the beneficial clinical response obtained with small doses of prednisone, suggest that both the cold-reactive antibodies and the IgG-kappa on circulating red cells were pathophysiologically significant. This is the first report of a patient with multiple red cell autoantibodies in whom serum complement component titers were determined in conjunction with characterization of the anti-red cell immunoglobulins. Subclinical infectious mononucleosis may have preceded the prolonged hemolytic episode. Clinical evidence of systemic lupus erythematosus has not appeared.


Anosmia and hypogonadism with ovarian mosaicism. An 18-year-old woman with anosmia and hypogonadotropic hypogonadism is presented. In addition endocrinologic evaluation revealed an apparent deficiency in pituitary growth hormone secretion in response to hypoglycemia and an ovarian insensitivity to exogenous gonadotropins. The ovaries, which on histologic examination appeared to be normal, upon karyotyping showed a chromosomal mosaicism, probably 46, XX/47, XXF+.


Response of active transport of ions and spontaneous water flux to osmotic gradients in gastric mucosa. The effects of symmetric changes of the mucosal and serosal bathing solution on cell water content, net ion flux, and net water movement were studied in the isolated frog gastric mucosa. Similar to transmucosal concentration gradients that induce water movement and changes in cell water content, symmetric osmolality changes of the bathing solutions also produce changes in these functional parameters. Thus, increments from 165 to 286 mosmol/kg water in the osmolality of both solutions reduce cell water content from 2.37 plus or minus 0.12 to 1.30 plus or minus 0.20 ml/g wt, the net ion flux (acid secretion plus short-circuit current) from 4.83 plus or minus 0.36 to 3.44 plus or minus 0.26 mueq/cm2 per h, and the net water flux from 10.6 plus or minus 1.1 to 2.4 plus or minus 1.2 mul/cm2 per h. These osmotically induced flux changes of water and ions must be considered when osmotic gradients are being used to generate and to evaluate water movement across the gastric mucosa.


Chloride transport through the n onshort-circuited isolated skin of Rana esculenta. The effects of acetazolamide (Diamox), ouabain, and amiloride on chloride movements through the isolated, nonshort-circuited Rana esculenta skin were investigated at different external NaCl concentrations. From hyposmotic solutions of low NaCl concentration (2 mM), two principal components of the chloride uptake may be distinguished: one dependent on the osmotic water flux and the other inhibited by Diamox but not by ouabain, amiloride, or removal of sodium from the external solution. Chloride uptake from a Ringer solution is always inhibited when either the sodium transport is blocked with amiloride or ouabain or when sodium is absent from the external solution. It can be concluded that the predominant component of the chloride transport mechanism when the external solution is concentrated (Ringer) is a sodium-dependent one. When chloride is absorbed from a solution of low concentration, however, a component predominated which is independent of sodium transport. It is suggested that the latter mechanism should be capable of exchanging chloride against endogenous base as in the in vivo skin.


Two-year follow-up of angina pectoris: medical or surgical therapy. The effects of bypass graft surgery versus continued medical management in 40 patients with stable angina were evaluated at 2 years: 17 of 20 surgical patients (85%) and 18 of 20 medical patients (90%) were alive, and 5 of 20 surgical patients (25%) and 2 of 20 medical patients (10%) had developed myocardial infarction; 8 of 17 surgical patients (47%) and 4 of 18 medical patients (22%) had no angina, and 13 of 17 surgical patients (76%) and 9 of 18 medical patients (50%) had no angina or greater than 25% increase in exercise time until angina. No statistically significant differences were demonstrated between the medical and surgical groups for all variables submitted to statistical analysis.


Adult toxoplasmosis presenting as polymyositis and cerebellar ataxia. A patient with toxoplasmosis had cerebellar, spinal cord, nerve root, and skeletal muscle symptoms. Toxoplasma gondii was isolated from skeletal muscle, and toxoplasma antibody titers were persistently elevated. The disease has been recurrent during a 3-year period with only temporary response to specific antimicrobial therapy.


Dermatomyositis in six patients without initial muscle involvement. Six patients initially had the characteristic skin eruption of dermatomyositis but no other evidence of the disease. They all later developed dermatomyositis. The clinical, laboratory, and pathological data suggest that whereas polymyositis may occur without skin involvement, the cutaneous features od dermatomyositis may be seen in the absence of detectable muscle disease. This variant of dermatomyositis occurs in children and adults but has not been found to be associated with internal malignant disease. The persistence of skin lesions despite systemic corticosteroid therapy is a poor prognostic sign. There were two patients with pulmonary fibrosis and dermatomyositis in this group. The presence of pulmonary fibrosis can contribute to a delay in diagnosis of dermatomyositis. Patients with pulmonary fibrosis had a poorer prognosis, despite moderate dosages of corticosteroids.


"No-vent" technique of coronary artery bypass. Isolated coronary artery bypass grafting was performed in 100 patients without the use of a left ventricular vent. Distention of the left ventricle did not occur. There was 1 late death unrelated to this technique. Vents in the left heart are not required under most circumstances.


A new application of counterpulsation: safer laparotomy after recent myocardial infarction. A 56-year old man suffered severe gastrointestinal tract bleeding five days following an anterior myocardial infraction. Nonoperative treatment consisting of gastric lavage, selective angiographic procedures on the gastric arteries with intra-arterial vasopressin injection (Pitressin), and massive blood replacement over a five-day period failed to halt hemorrhage. Institution of intra-aortic balloon pumping reversed the patient's hemodynamic deterioration and made possible a definitive operation. The successful outcome in this patient suggests that counterpulsation may play a useful role in lowering the formidable surgical mortality observed in patients with acute myocardial injury who require emergency noncardiac operations.


Multiple primary malignant neoplasms. A search for an immunogenetic basis. The occurrence of multiple primary malignant neoplasms in single individuals is well documented. Although many hypotheses have been advanced to explain this occurrence, there has been no study to determine if a presumed "increased susceptibility to cancer" has an immunogenetic basis. We evaluated the cellular immunity and histocompatibility antigens of 42 patients who had had from two to four multiple primary malignant neoplasms. We failed to demonstrate a preexisting impairment of immunocompetence or abnormal HL-A antigen frequencies in these patients. The occurrence of multiple primary malignant neoplasms in related tissues, eg, lung/larynx/oral cavity, and the occurrence of successive primary malignant neoplasms at a time interval consistent with the patient's being cured of preceding malignant neoplasms suggest that multiple primary malignant neoplasms result from repetitive induction by the same or similar etiologic factors in patients who are cured after treatment of the first malignant neoplasm.


Supraceliac pseudoaneurysm of abdominal aorta. Staged repair employing preliminary axillary-femoral bypass. The fifth successfully treated pseudoaneurysm of the visceral branch segment of abdominal aorta emphasizes individualized management of such complex lesions. Staged operative management of a supraceliac pseudoaneurysm by preliminary axillary-femoral bypass minimized potential intraoperative hazards. Visceral protection during aortic occlusion was simplified by axillary-femoral perfusion.


Emergency and urgent operations for ulcerative colitis. The procedure of choice. Eighty consecutive emergency and urgent colectomies for ulcerative colitis were performed. One-stage total proctocolectomy was performed in 37 patients, with a 9.1% mortality; ileostomy with subtotal colectomy was performed in 43, with a 7.0% mortality. The overall mortality was 7.5%. Postoperative morbidity after total proctocolectomy (mean postoperative hospitalization, 27.6 days; nonseptic complication rate, 29.4%; septic complication rate, 29.4%) was not substantially different from that after subtotal colectomy (postoperative hospitalization, 33.3 days; nonseptic complications, 45.0%; septic complications, 35.0%). Survivors of subtotal colectomy required abdominal-perineal resection of the colorectal remnant in 75.7% of patients, and no patient had successful subsequent ileorectal anastomosis. It is suggested that one-stage total proctocolectomy be adopted as the surgical procedure of choice in emergency or urgent operations for ulcerative colitis.


Superficial microcystic corneal dystrophy. Microcystic corneal dystrophy was first described in 1964. Since then, several reports have elaborated on the original findings plus other associated corneal changes. The cause is unknown, but pathological studies have been done in several cases. The corneal changes are subtle and easy to miss on routine examination. This study of 17 patients was undertaken to further evaluate the signs and symptoms of this abnormality and to emphasize techniques of examination. Detailed clinical and laboratory evaluations of tear function were performed. No definite association between tear function and the corneal changes could be determined. In this group of patients, the process seemed benign with minimal symptoms.


Dissecting aneurysms of coronary arteries. A case of dissecting aneurysms of coronary arteries caused by cardiac surgery was found. Review discloses that most coronary arteries arise spontaneously, occur more often in women than men, frequently in the postpartum period, and are characterized by the sudden onset of acute chest pain in a previously well middle aged patient who abruptly collapses and dies. The anterior descending branch of the left coronary artery is most frequently affected, the dissection usually involves the first portion of that single vessel, and the lesion appears amenable to bypass venous grafting. While the cause is unknown, the frequency of pregnancy, the loosening effect of that condition on connective tissue, and the occasional finding of medial degeneration of the arterial wall suggest cystic medial necrosis as an important etiologic consideration. This case report recommends gentle handling of coronary arteries in surgical procedures.


Influence of head orientation on visually induced pitch and roll sensation. Observers viewing rotating scenes in their periphery frequently experience self-motion in the opposite direction. A full field (360 degrees) flight simulator projection system was used to investigate the sensations resulting from pitch, roll, and yaw stimuli at various head orientations. Steady yaw rate (circularvection) and development of a constant roll tilt angle, for the head erect and constant velocity yaw and roll stimuli, confirmed previous reports. Pitch stimuli also were found to produce a sensation of tilting to a steady pitch angle, which was much stronger for pitch forward than backward. Pitch and roll effects were strongly dependent on head position, increasing for the head rolled 90 degrees to the side or inverted, and decreasing for the head pitched 25 degrees forward. These results support a hypothesis that visually induced tilt is limited by conflict with otolith information.


Abnormalities of factor VIII and platelet aggregation--use of ristocetin in diagnosing the von Willebrand syndrome. Ristocetin was used to study platelet aggregation in platelet-rich plasma and to assay the von Willebrand factor activity of factor VIII (VIII-VWF). Ristocetin-induced platelet aggregation (RIPA) was decreased in 13 of 18 patients with von Willebrand's disease (VWD) who had decreased plasma levels of VIII-VWF. The five patients with normal RIPA appeared to have mild VWD but did not constitute a separate subclass. RIPA was also abnormal in some patients with intrinsic platelet defects, but in no case was the defect corrected by normal plasma. The latter type of correction appears to be specific for VWD. Aspirin ingestion inhibited the second phase of RIPA (at low concentrations of ristocetin only) but did not affect the initial phase of aggregation or the level of VIII-VWF. We also studied a group of patients who had both abnormalities of the factor VIII complex and intrinsic platelet defects, such as impaired collagen-induced aggregation, as well. The findings in these patients and in those with typical von Willebrand's disease appear to comprise a spectrum of disorders (the von Willebrand syndrome) in which some abnormality of the factor VIII complex is associated with impaired platelet function. At present, ristocetin would appear to be a useful reagent for evaluating patients with bleeding disorders and for studying patients with the von Willebrand syndrome.


Rheumatoid nodulosis: an unusual variant of rheumatoid disease. A middle-aged man developed multiple subcutaneous nodules associated with palindromic rheumatism. There was little evidence of synovitis; however multiple cyst-like intraosseous radiolucencies were noted. Nodules from two sites were histologically typical rheumatoid nodules. Subjective and objective improvement occurred during penicillamine therapy. This clincial presentation seems sufficiently distinctive to warrant characterization as a variant of rheumatoid disease termed by us: rheumatoid nodulosis.


Abdominal approach for the ligation of bleeding oesophageal varices. The conventional surgical treatment of bleeding oesophageal varices in the emergency situation is based upon the Boerema-Crile operation of transthoracic oesophagotomy and ligation of the varices. This and the other methods of treatment of this condition in current practice are discussed. Two cases are reported in which a transabdominal oesophagogastrotomy was used to approach the site of bleeding. This operation is described and the theoretical and practical advantages it appears to offer over the standard approach are considered.


The effect of cyclophosphamide on B and T lymphocytes in patients with connective tissue diseases. Absolute numbers of B (IgG-, IgM-, and IgA-staining) and T lymphocytes (sheep erythrocyte rosette-forming cells) were determined in patients with systemic lupus erythematosus and other connective tissue diseases in cyclophosphamide-treated and noncyclophosphamide-treated patients and in control subjects. In patients receiving cyclophosphamide, all three types of immunoglobulin-staining varieties of circulating B lymphocytes were significantly decreased. At the same time the circulating T lymphocytes were also significantly reduced. In patients with scleroderma treated with therapeutic doses of cyclophosphamide and studied sequentially, the reduction in B lymphocytes occurred first, with eventual depletion of both cell types. In 2 patients, an early rebound increase in T cells occurred followed by a marked reduction. These data indicate that the immunosuppressive effects of cyclophosphamide may be associated with a reduction in both cell types.


Pericarditis, pleural effusion, and pneumonitis with transient mitochondrial antibodies. Four women with fever, arthromyalgias, pericarditis, pleural effusion, high erythrocyte sedimentation rates, and lymphopenia had mitochondrial antibodies in the serum in the absence of antinuclear antibody. Their illness lasted 5-12 weeks and the antibody test results became negative on remission. Absence of specific bacteriological findings, normal antistreptolysin O titres, resistance to antibiotics, and good response to steroids suggested that these cases represented a milder and less persistent form of the syndrome resembling systemic lupus erythematosus described by German authors.


Hemophilus parainfluenzae meningitis.

Hemophilus parainfluenzae meningitis. Although Hemophilus influenzae is a common cause of meningitis, other members of the Hemophilus genus are rarely the infecting organism. Of 56 cases of meningitis due to Hemophilus species obseved at one hospital in the period 1970-74, 53 were due to H. influenzae and 3 to H. parainfluenzae. In the cases of H. parainfluenzae meningitis the clinical picture was complicated by associated sepsis, and therapy with ampicillin was not entirely satisfactory.


Suppression of secondary cellular immunity to a tumor allograft by cyclophosphamide and 1,3-bis(2-chloroethyl)-1-nitrosourea. C57BL/6 mice (H-2b) were immunized with lethally x-irradiated Moloney virus-induced lymphoma cells of BALB/c origin (H-2d) on Days 0 and 10 and received rug on Days 11 and 14. Their spleen cells were then tested for reactivity against Moloney virus-induced lymphoma of BALB/c origin by the 51Cr-release cytotoxicity assay. In non-drug-treated mice the secondary cytotoxic response was maximal on Days 14 to 15, declined rapidley, and recurred after Day 21. The cytotoxic effector cells were shown to be theta-bearing T-lymphocytes. Cyclophosphamide (CY), 180 mg/kg, given on Day 11, totally prevented the development of a cytotoxic response and when given on Day 14 abolished the response already established. CY, 48 mg/kg, as well as 1,3-bis(2-chloroethyl)-1-nitrosourea 33 mg/kg, were almost as suppressive. Immune mice given CY on Day 14 and reimmunized on Day 36 exhibited a normal tertiary response. Mice similarly immunized on Days 1 and 10 and given drugs on Day 14 were challenged on Day 15 with up to 3.5 x 10-8 viable Moloney virus-induced lymphoma cells of BALB/c origin. Despite H-2 incompatibility, all nonimmune control mice developed ascites and died, whereas all mice immunized but not given drug failed to develop ascites. By contrast, 17 of 34 immunized mice given CY, 180 mg/kg, and 7 of 34 given 1,3-bis(2-chloroethyl)-1-nitrosourea developed ascites. The ascites eventually regressed. THE RESULTS SHOW THAT CY and 1,3-bis(2-chloroethyl)-1-nitrosourea can suppress a secondary cellular immune response as measured by the T-cell-mediated 51Cr-release cytotoxicity assay in vitro and by viable tumor challenge in vivo.


Metastasis formation of Yoshida sarcoma heterotransplanted in adult golden hamsters treated with anti-hamster thymocyte serum. Growth of Yoshida sarcoma cells heterotransplanted in adult golden hamsters treated with antithymocyte serum was investigated. Control animals were grafted with or without hydrocoritisone conditioning. In the antithymocyte serum-treated group, tumors were produced in cheek pouches and grew progressively in size until Day 21 with no sign of regression. Little host reaction was noticed histologically around the tumor grafts throughout the course. Distant metastases were revealed at autopsy in 9 of 12 animals from Day 14 of inoculation on, and all the experimental animals died by Day 23. Six of the 9 hamsters with metastases died of tumor. By means of the back-transplantation test, the metastatic tumor cells were shown to be consistent with Yoshida sarcoma cells. In hydrocortisone-treated and untreated groups, the tumor size reached a maximum on Day 21 and Day 5, respectively, and then regression followed. No metastasis was detected in either control group.


Malignant transformation of mouse cells by cigarette smoke condensate. A low-nicotine cigarette smoke condensate, 12 fractions of the condensate, and a reconstituted sample were tested for their ability to induce transformation in the mouse cell line 3H/10T-1/2 CL-8. This cell line is noted for its remarkable low spontaneous rate of transformation. Both the crude condensate and the reconstituted sample as well as two specific fractions induced transformation in the mouse cells. These transformed cells produced fibrosarcomas when injected s.c. into antithymocyte serum-treated syngeneic mice.


Changes in RNA in relation to growth of the fibroblast. III. Posttranscriptional regulation of mRNA formation in resting and growing cells. The proportion of hnRNA processed into cytoplasmic mRNA in resting and growing 3T3 and 3T6 cells has been investigated by measuring the efficiency of transfer of nuclear poly(A) to the cytoplasm. When nuclear poly(A) is either labeled continuously with precursor or pulse labeled and then chased with cordycepin, growing cells transfer approximately twice as much of the poly(A) from the nucleus to the cytoplasm as do resting cells. When cells undergo a serum-induced transition from the resting to growing state, the efficiency of poly(A) transfer is increased to that characteristic of growing cells by 3 hr after the addition of the serum. On the other hand, the proportion of hnRNA which is polyadenylated and the total nuclear poly(A) content are the same in resting and growing cells. It appears that the principal means by which the cell regulates its poly(A)+ mRNA content in the growing and resting states is by regulating the efficiency which nuclear poly(A)+ RNA is converted to cytoplasmic mRNA.


Effect of alterations of arterial blood pressure and heart rate on segmental dyskinesis during acute myocardial ischemia and following coronary reperfusion. The effect of alterations of blood pressure and heart rate on segmental dyskinesis induced by coronary artery ligation and the influence of such manipulations on the magnitude of recovery of the syskinetic segment during subsequent coronary artery reperfusion were evaluated in 77 open-chest dogs. Wall motion was recorded by ultrasound reflected directly from the ischemic myocardial segment. Acute ischemia produced characteristic regional abnormalities in wall motion: aneurysmal bulging occurred during isometric contraction and wall velocity was markedly reduced during ventricular ejection. During 69 minutes of ischemia, a control group of dogs underwent no interventions and showed no further changes in wall motion. Tachycardia induced by atrial pacing during ischemia had no significant effect. Arterial hypertension during ischemia caused a marked reduction in wall velocity when methoxamine was used: 14 plus or minus 2 (SE) mm/sec (ischemia alone) to 6 plus or minus 1 mm/sec (ischemia + drug). In contrast, norepinephrine improved wall velocity: 11 plus or minus 2 mm/sec (ischemia alone) to 25 plus or minus 4 mm/sec (ischemia + drug). Hypertension caused by infusion of phenylephrine gave intermediate results, as did hypotension induced by either nitroprusside or hemorrhage during the ischemic period. After 60 minutes the drugs were stopped, the coronary ligation released, and the ischemic myocardium reperfused. The relative order of improvement of wall velocity with reperfusion was 11 plus or minus 2 mm/sec (ischemia alone) to 24 plus or minus 3 mm/sec (reperfusion) in the group that received norepinephrine, 12 plus or minus 3 mm/sec to 20 plus or minus 3 mm/sec in the control group with no intervention durin ischemia, 13 plus or minus 2 mm/sec to 20 plus or minus 1 mm/sec in the nitroprusside group, 9 plus or minus 2 mm/sec in the phenylephrine group, and 14 plus or minus 2 mm/sec to 12 plus or minus 1 mm/sec in the methoxamine group. The aneurysmal bulging during isometric contraction  also was reduced to a greater degree by reperfusion in the group that received norepinephrine during the ischemic period than it was in the groups undergoing other interventions during ischemia. We conclude that drug-induced elevations in arterial blood pressure can have different effects on the syskinetic motion of acutely ischemic myocardium and on the degree of recovery following reperfusion depending on the particular agent used.


Demonstration that the atria, ventricles, and lungs each are responsible for a tonic inhibition of the vasomotorcenter in the dog. To localize the areas of the cardiopulmonary region involved in tonic inhibition of the vasomotor center, anesthetized dogs were subjected to sinoaortic denervation and diaphragmatic vagotomy. Afferent vagal nerve traffic was interrupted in the neck by cooling. With the venous return taken from the venae cavae, oxygenated extracorporeally, and returned to the aorta, the heart was removed, leaving the ventilated lungs (condition 1), and the lungs and the ventricles were removed, leaving the beating atria (condition 3). With the venous return taken from the pulmonary arteries, oxygenated extracorporeally, and returned to the left atrium, the lungs were removed, leaving the intact working heart (condition 2), and the lungs were removed and the atria were denervated, leaving the working innervated ventricles (condition 4). Vagal cooling increased aortic pressure by 25 plus or minus 2 (SE) mm Hg in condition 1, by 36 plus or minus 2 mm Hg in condition 2, by 29 plus or minus 2 mm Hg in condition 3, and by 29 plus or minus 7 mm Hg in condition 4. Removing the atria in condition 3 or denervating the ventricles in condition 4 abolished the reflex response. Thus, afferent vagal nerves from the lungs and the heart tonically inhibit the vasomotor center. The inhibition exerted by the heart is caused by receptors in the atria and the ventricles.


Chronic polymicrobial bacteremia.

Chronic polymicrobial bacteremia. A 13-year-old girl had chronic polymicrobial bacteremia with Enterobacter hafnia and Enterobacter agglomerans in the absence of any demonstrable underlying illness, use of immunosuppressive drugs, or discovery of portal of entry of the bacteria. She was treated successfully with a prolonged course of carbenicillin and aminoglycoside antibiotics which were tolerated well. The only side effect of the therapy was a transient episode of vestibular dysfunction which was reversible following cessation of gentamicin. The principles of management of polymicrobial bacteremia are presented.


[Selective coronary perfusion scintigraphy. II. Abnormal coronary perfusion pattern (author's transl)]. Integrated assessment of selective coronary angiorgrams and perfusion scintigrams of 86 patients with coronary-artery insufficiency revealed anatomical and topographical details which could not be achieved with one of these methods alone. Pathological perfusion patterns were quite different from normal ones. The functional effects of single, centrally located stenoses, of more peripheral and multiple narrowings, as well as of anastomoses and collaterals were adequately demonstrated by selective coronary perfusion scintigraphy and prevented diagnostic errors. It is, therefore, an additional aid in the diagnosis of coronary-artery disease.


Portacaval anastomosis and peptic ulcer: a nonassociation. The incidence of peptic ulcer is increased in cirrhosis and is widely believed to be even greater in cirrhotic patients with portacaval anastomosis (PCA). Two prospective, controlled investigations of prophylactic PCA were evaluated to compare the frequency of peptic ulcer in two groups of cirrhotic patients with similar clinical and laboratory manifestations of cirrhosis randomly selected to be an unoperated Control Group (60 patients) or to have PCA (Shunt Group, 48 patients). In addition, nonrandomized groups of cirrhotic patients, 77 of whom were excluded from the randomized study and 44 of whom had therapeutic PCA, were studied. A diagnosis of chronic peptic ulcer was based on the demonstration of an ulcer crater by X-ray, endoscopy, surgery, or autopsy. Prior to inclusion in these studies, approximately 10% of patients had had peptic ulcer. After inclusion, during a mean follow-up period of 45 months, 12% of both the Control and Shunt Groups developed peptic ulcers. The frequency of complications of peptic ulcer, of recurrence of peptic ulcer, or of acute or symptomatic (unproved) ulcer were similar in both groups. Ulcers tended to develop later in shunted than in unshunted patients. Similar data were obtained from three of four other controlled investigations of PCA. This investigation does not find an increased occurrence of peptic ulcer after PCA. The frequency of ulcer in cirrhosis appears to increase with the duration of the disease independent of the presence or absence of PCA.


Role of bile acids in the pathogenesis of aspirin-induced gastric mucosal hemorrhage in rats. The simultaneous oral administration of acetylsalicylic acid (64 mg per kg) and taurocholic acid (2.5 mM) to rats induced gastric hemorrhagic mucosal bleeding in 82.4% of animals compared with 29.7% of animals given acetylsalicylic acid alone. Taurodeoxycholic acid and glycocholic acid with acetylsalicylic acid also significantly increased the incidence of gastric hemorrhagic mucosal bleeding compared with the control animals, but cholic acid, chenodeoxycholic acid, and deoxycholic acid did not have a significant effect. Hemorrhagic mucosal lesions were not observed when the bile acids studied were administered alone. These results indicate a possible role for certain bile acids in the pathogenesis of gastric mucosal bleeding after the ingestion of aspirin.


Demonstration of superficial gastric erosions by double contrast radiography. Superficial gastric erosions were found in 30 of 267 patients undergoing esophagogastroduodenoscopy. In 13 of these patients, and in 7 others not submitted to endoscopy, superficial gastric erosions were demonstrated radiologically by the double contrast technique. These may appear as flat, linear streaks, as dots of barium, or as target-like lesions with a central fleck of barium surrounded by a radiolucent halo. In 1 patient, a ring-shaped density was demonstrated due to a small blood clot adherent to a gastric erosion. Although patients presenting with acute hemorrhagic gastritis should still be examined first by endoscopy, double contrast radiography has been of value in patients presenting after the cessation of bleeding and in patients with vague digestive complaints. Because of our experience with the diagnosis of standard methods, and because of the Japanese experience with diagnosis of superficial gastric carcinoma, it is recommended that this technique be applied more widely to facilitate the detection of early and small lesions in the stomach.


Enhancement of reaginic antibody formation in the mouse by concanavalin A. SWR mice injected with Concanavalin A (Con A) and Ovalbumin (Ov) one immediately after the other or with mixtures of Con A and Ov produced reaginic antibodies to Ov (titer 1:80) after primary immunization and reaginic antibody levels of 1:640 after secondary immunization. Mixtures of Con A and Ov were shown to contain soluble complexes which could be precipitated by 20% polyethylene glycol (PEG). Mixtures of Con A and human serum albumin or Con A and bovine serum albumin did not form soluble complexes and did not induce detectable reagin formation. The possible role of soluble of Con A-Ov complexes in stimulating reaginic antibody is discussed.


Biological relevance of alkali-labile sites in double-stranded DNA after gamma-irradiation. Alkali-labile sites produced by gamma-irradiation in dry RF-DNA of bacteriophage phi chi 174 were found to be lethal lesions with respect to the plaque-forming ability.


Effects of gamma-radiation on the optical properties of aqueous lysozyme-polyinosinic-acid complexes. The absorption of gamma-rays by dilute aqueous solutions of lysozyme and polyinosinic acid (poly I) causes significant increases in the light-scattering power of room temperature solutions in certain critical concentration regions. Light-scattering properties are unaltered by relatively large doses of radiation delivered to heated complexes. Prior irradiation of the poly I alone yields complexes whose light-scattering properties are the same as those of the unirradiated system. Studies of the radiation-induced absorbance changes of these complexes at 260 and 280 nm show behaviour indicative of the radiation chemistry of polynucleotide rather than of lysozyme. From the evidence available, it is postulated that most of the damage is to the poly I rather than lysozyme in the complexes. Increases in light-scattering and dissymmetry are attributed to unwinding of the multi-stranded polynucleotide. The role of water in the formation and radiation sensitivity of the complexes is discussed.


Haemopoietic stem-cell kinetics during continuous irradiation. Analysis of the kinetics of the biphasic response of haemopoietic stem cells to chronic low dose-rate irradiation indicates that a highly radiosensitive delay of entry into cell-cycle by Go cells may be responsible for the initial, apparently dose-rate-independent, phase of the dose-response curve. The second component of the curve appears to be the result of a combination of two factors: a relatively decreased differentiation demand on stem cells due to increased amplification in the transit-committed precursor cells, and a uniform shortening of the cell-cycle--including the S period--in the stem cells.


DNA-to-protein cross-linking in synchronized HeLa cells exposed to ultra-violet light. The U.V.-induced DNA-to-protein cross-linking in synchronously growing HeLa cells exhibits age-dependent variations. The maximum yield of cross-linking is observed during S-phase and exposure of mid-S cells to 250 ergs/mm2 renders about 22 per cent of DNA unextractable owing to cross-linking with proteins. Cells irradiated in G1 or G2 exhibit much smaller amount of cross-linking. The unextracted DNA was recovered from the protein fraction, and the addition of 0-5 per cent trypsin solution rendered all the DNA extractable. Fluctuations in the age-response pattern for the yield of DNA-protein cross-links varies throughout the cell-cycle in the same way as the efficiency for cell-killing, i.e. cells are resistant in G1, sensitive as they move into and through the S phase, and resistant again in G2. Dose-response curves determined at different times in the cycle reveal that up to about 500 ergs/mm2 the amount of DNA recovered from S cells decreased for about 8 per cent for every 100 ergs/mm2 of U.V. light. The amount of recovered DNA remained unchanged for cells exposed to 500 ergs/mm2 in the middle of S, for as long as 8 hours after exposure, i.e. for as long as the experiments were carried out no recovery from this damage was observed.


A major component of the radiation effect: interference with endocellular control of cell proliferation and differentiation. Cytological investigations with diploid and autotetraploid plant materials reveal that cytoplasmic growth, including the propagation of autoreduplicating cytoplasmic units such as mitochondria and proplastids, continues during the radiation-induced mitotic delay, thus increasing the ratio of cytoplasmic amount: nuclear ploidy. In accordance with a control mechanism, detected in the plant endosperm, this increase causes mitotic inhibition due to any early induction of otherwise normal differentiation processes. Although mutations are not a prerequisite, they can act in an auxiliary manner by quantitative action. The contribution of 'early differentiation' to non-survival varies according to irradiation conditions and can outweigh the direct effects of irreversible genetic damage. Therefore, a higher level of ploidy cannot improve survival significantly, if mitotically-active tissues are irradiated under the usual conditions. Corresponding observations are known from animals. The results suggest that the control mechanism of the endosperm is (a) general to multicellular organisms, and (b) can be modulated by extracellular agents.


Interaction of gamma-ray and neutron effects on the proliferative capacity of Chinese hamster cells. The loss of reproductive capacity of Chinese hamster cells has been studied using 60Co gamma-rays and DT neutrons singly or combined in fixed proportions. Surviving fractions were measured down to 2 times 10-4. Interaction between the effects produced by neutrons and gamma-rays in the mixed irradiations has been demonstrated. The results are compared with those predicted by the cellular inactivation model developed by Katz and co-workers from their delta-ray theory of track structure.


Calcium-containing structures in vertebrate glial cells. Ultrastructural and microprobe analysis. Electron probe microanalysis has revealed that vesicular or cisternal structures containing electron-dense material in frog ependymal glial cells contain deposits of calcium and phosphorus. The so-called "osmiophilic particles" in human astrocytes also contain calcium. It is suggested that these organelles are storage sites of calcium.


Electrophoretic analysis of liver and testis histones of the frog Rana pipiens. Histones were extracted from frog livers and testes and analyzed by electrophoresis on long polyacrylamide gels and on sodium dodecyl sulfate (SDS)-containing polyacrylamide gels. Frog histones were found to be similar to those of calf thymus except that frog histone fraction F2A2 showed a marked dependence on the temperature at which the long gels were run, and frog histone fraction F3 could be separated from frog F2B on SDS-containing gels. Comparisons between frog liver and frog testis histones indicated that the testis contains as its major F1 component a fast migrating species not found in liver. Testis histones also showed less microheterogeneity of fractions F3 and F2A1 than liver histones. These were the only differences observed between liver and testis histones, even when testis histones were prepared from sperm suspensions that were rich in cells in the late stages of spermiogenesis. Thus it seems that, in Rana, the electrophoretic properties of the basic proteins of sperm differ from those of somatic cells only in the nature of histone F1 and in the degree of microheterogeneity of fractions F2A1 and F3.


Size and shape of transverse tubule openings in frog twitch muscle fibers. The openings of transverse tubules in frog twitch fibers are described. The tubules open to the extracellular space by a narrow neck, with an inner diameter of 20 nm. The most peripheral portion of the tubules is tortuous and has a variable diameter. The similarity in size of the openings of T tubules and caveolae and the meandering path of the tubules are sufficient to account for the paucity of observed openings.


Complement fixation by rheumatoid factor. The capacity for fixation and activation of hemolytic complement by polyclonal IgM rheumatoid factors (RF) isolated from sera of patients with rheumatoid arthritis and monoclonal IgM-RF isolated from the cryoprecipitates of patients with IgM-IgG mixed cryoglobulinemia was examined. RF mixed with aggregated, reduced, and alkylated human IgG (Agg-R/A-IgG) in the fluid phase failed to significantly reduce the level of total hemolytic complement, CH50, or of individual complement components, C1, C2, C3, and C5. However, sheep erythrocytes (SRC) coated with Agg-R/A-IgG or with reduced and alkylated rabbit IgG anti-SRC antibody were hemolyzed by complement in the presence of polyclonal IgM-RF. Human and guinea pig complement worked equally well. The degree of hemolysis was in direct proportion to the hemagglutination titer of the RF against the same coated cells. Monoclonal IgM-RF, normal human IgM, and purified Waldenström macroglobulins without antiglobulin activity were all inert. Hemolysis of coated SRC by RF and complement was inhibited by prior treatment of the complement source with chelating agents, hydrazine, cobra venom factor, specific antisera to C1q, CR, C5, C6, or C8, or by heating at 56 degrees C for 30 min. Purified radiolabeled C4, C3, and C8 included in the complement source were bound to hemolysed SRC in direct proportion to the degree of hemolysis. These data indicate that polyclonal IgM-RF fix and activate complement via the classic pathway. The system described for assessing complement fixation by isolated RF is readily adaptable to use with whole human serum.


Osteogenic sarcoma. Immunologic parameters before and during immunotherapy with tumor-specific transfer factor. 18 patients with osteogenic sarcoma were followed by serial measurements in vitro of tumor-specific cell-mediated cytotoxicity and of "active" and total rosette-forming T-cells. 13 of these patients have had or are currently receiving injections of osteogenic sarcoma-specific dialyzable transfer factor derived from healthy donors. In three patients with very small lesions, cytotoxicity was high before amputation and decreased within 2 mo after removal of tumor. Cytotoxicity was low at time of diagnosis in all patients with large tumor masses. The cytotoxicity of the patients' lymphocytes increased after administration of tumor-specific transfer factor in all patients so treated. Patients receiving nonspecific transfer factor showed evidence of declining cell-mediated cytotoxicity. Tumor-specific transfer factor may produce an increase in cell-mediated cytotoxicity to the tumor in patients with osteogenic sarcoma. This possibility is suggested by the pain and edema that occurred in the area of the tumor in patients who had metastatic disease when therapy was started and by lymphocytic infiltrates in the tumor, as well as by the increase in cell-mediated cytotoxicity and the increase in percentage of active rosette-forming cells from subnormal to normal. Serial measurements of cell-mediated cytotoxicity are helpful in monitoring the efficacy of transfer factor and other modes of therapy in these patients, and these measurements are the best available criteria for selection of donors of tumor-specific transfer factor.


Tumor-specific cell-mediated immunity in household contacts of cancer patients.. Patients with osteogenic sarcoma (and related tumors), hypernephroma, and breast carcinoma, and their household contacts were tested for tumor-specific cell-mediated immunity against these tumors with the use of a short-term chromium-51 release assay. This assay, reproducible over many months and well-correlated with the clinical course of the patients, was used to demonstrate that household contacts of patients with osteogenic sarcoma and breast carcinoma have specific immunity against the tumor type with which they have been in contact. In both types of tumors, the range of cytotoxicity values produced by lymphocytes from the household contacts was significantly higher than that of the normal population. The incidence of immunity was much higher in household contacts of patients with breast carcinoma than in those of patients with osteogenic sarcoma. Immunity was found with equal frequency in men and women, as well as in genetically and nongenetically related household contacts (guardians, adopted children, spouses). Immunity against hypernephroma was not demonstrated in either patients with hypernephroma or their household contacts.


A cytofluorometric and radiochemical analysis of the uptake and turnover of 5-hydroxytryptamine in mast cells. Uptake and turnover of 5-hydroxytryptamine (5-HT) after the administration of 5-hydroxytryptophan (5-HTP) has been studied in rat peritoneal mast cells using radiochemical and quantitative cytofluorometric methods. There was a close agreement between the results obtained with the two methods. The extreme sensitivity of the cytofluorometric method was indicated by the fact that 0.2 pg 5-HT contained in mast cells from control rats could be readily quantitated. The cytofluorometric analysis demonstrated a large variation in 5-HT content of individual mast cells within the mast cell populations. The storage capacity for 5-HT greatly exceeded the amount found in normal mast cells. Intraperitoncally injected 5-HTP was rapidly taken up and eliminated from the mast cells within 12 hr, while 5-HT, probably derived from intracellular decarboxylation of 5-HTP, was retained in the cells and slowly eliminated. The elimination of 5-HT followed an exponential course and the half-life was found to be about 10 days. The results indicate that the turnover of 5-HT is much slower in mast cells than in other cells which normally store 5-HT.


Effects of bacterial lipopolysaccharide on the induction and expression of cell-mediated immunity. II. Stimulation of the efferent arc. The delayed-type hypersensitivity (DTH) produced by subcutaneous or intravenous immunization of mice with sheep red blood cells (SRBC) is mediated by T cells in cooperation with accessory cells (monocytes). Different effects of bacterial lipopolysaccharide (LPS) on this T cell activity were found at different stages of the immune response. a) T cell activity was enhanced by LPS given 1 to 3 days after sensitization. Enhancement, which was most marked when treatment was given on the 2nd day of the immune response, did not occur unless LPS was introduced into a site that drains to the responding lymph node. This stimulatory effect was dose dependent. b) A similar effect on the response to intravenous immunization caused DTH to appear transiently in mice given a T cell-blocking dose (10-9) of SRBC. c) DTH was suppressed by LPS given at the height of the T cell response (day 4). This effect, which was short lived and reversible, was due to the action of LPS on the accessor cells needed for the expression of DTH. Treatment of recipients with LPS completely blocked the adoptive transfer of DTH to both normal and spenectomized mice. The ability to react was regained slowly, but it returned promptly to splenectomized recipients. LPS has clearly more than one effect on the expression of cell-mediated immunity because it affects specific and non-specific mediator cells in different ways.


A lectin model for unresponsiveness. I. Reversible effects of high doses of concanavalin A on spontaneous and induced DNA synthesis. A mitogen model for unresponsiveness has been studied. In 72-hr cultures the lowest supraoptimal dose of Concanavalin (Con A) showing no stimulation of rat thymocytes above background was 160 mug/ml. This dose was also found to inhibit substantially (50%) the high spontaneous DNA synthesis by thymocytes during the first 24 hr of culture. This effect was most significant 12 to 24 hr after addition of high-dose Con A. It was not necessary for the cells to remain continuously with high-dose Con A, since washing out unbound Con A after 1 hr still led to reduced 3-H-thymidine incorporation at 24 or 72 hr. Furthermore, high-dose Con A treatment suppressed DNA synthesis when added at 48 hr to cultured optimally stimulated with Con A. Interaction with high-dose Con A appears to inhibit the ability of cells to initiate new rounds of DNA synthesis without blocking ongoing DNA replication. Removal of Con A with alpha-methyl-D-mannopyranoside (mannose) led to only partial reversal of these effects after 1 hr although "unresponsiveness" was reversible at 15 min. Suppression was not due to cytotoxic or agglutinating effects of the high-dose Con A. Two times 10-6 thymocytes exposed to 160 mug of Con A for 1 hr bound 1.7 mug, 86% of which was elutable with mannose. There was no reduction in the ability of high-dose treated cells to bind a subsequent optimal dose of Con A. Since Con A binds to essentially all thymocytes, this system is useful in following early events in the induction of lectin "unresponsiveness" as a model for immunologic tolerance.


Characterization of immunoglobulin-bearing and other small lymphocytes in mouse bone marrow by sedimentation and electrophoresis. 125I-antiglobulin binding and radioautography have been used to define the sedimentation and electrophoretic properties of immunoglobulin-bearing and other small lymphocytes in bone marrow. Bone marrow cell suspensions from CBA mice were exposed to 125I-labeled rabbit anti-mouse immunoglobulin for 30 min at 0 degrees C. Cell fractions were collected after either sedimentation at unit gravity or continuous free-buffer film preparative electrophoresis. The sedimentation profiles of labeled, antiglobulin-binding small lymphocytes and of unlabeled small lymphocytes were identical, peaking at 2.6 mm/hr, except for 5% of the labeled small lymphocytes which sedimented at 3.8 to 4.1 mm/hr. In electrophoretic fractions most labeled small lymphocytes were contained in a peak of low mobility and unlabeled small lymphocytes were of intermediate mobility, but high mobility fractions also contained small numbers of labeled and unlabeled small lymphocytes.  Small lymphocytes comprised 80 to 85% of all nucleated bone marrow cells in fractions taken at the peak of the small lymphocyte distribution profile after either sedimentation or electrophoresis, accompanied mainly by either erythroid or granulocytic precursor cells, respectively. The results demonstrate that small lymphocytes with readily detectable surface immunoglobulin in the bone marrow closely resemble those in peripheral lymphoid tissues with respect to their sedimentation and electrophoretic properties. The possibility is raised that the rapidly sedimenting immunoglobulin-bearing small lymphocytes in bone marrow are a functionally distinct group of potential precursor cells. Non-antiglobulin-binding small lymphocytes in bone marrow resemble electrophoretically the double negative (immunoglobulin -and theta -negative) small lymphocytes in other lymphoid tissues. Small lymphocytes of high electrophoretic mobility, prominent in the theta- bearing lymphocyte populations of peripheral lymphoid tissues, are few in bone marrow. Lymphocyte-rich suspensions, containing either high or low proportions of immunoglobulin-bearing small lymphocytes, can be separated from bone marrow by electrophoresis


Antibody-mediated suppression of grafted lymphoma. III. Evaluation of the role of thymic function, non-thymus-derived lymphocytes, macrophages, platelets, and polymorphonuclear leukocytes in syngeneic and allogeneic hosts. Syngeneic or allogeneic mice pretreated with sublethal whole-body irradiation were rendered incapable of suppressing the growth of grafted tumor cells sensitized with alloantibody. The growth of sensitized tumor cells was suppressed when they were mixed with donor effector cells from mice syngeneic or allogeneic to the recipients and then were inoculated in irradiated recipients. Three donor-host combinations were used to study the suppression of the murine lymphoma 6C3HED indigenous to C3H mice. These were C3H donor cells in C3H recipients, C57BL/6 donor cells in C3H recipients, or C57BL/6 donor cells in C57BL/6 recipients. In all three combinations, macrophages obtained from an inflammatory exudate, exudate lymphocytes not bearing theta antigen, and platelets were, in descending order of effectiveness, consistently active in restoring antibody-mediated suppression of tumor growth in irradiated hosts. Prior irradiation of the transferred lymphocytes somewhat diminished their effectiveness. Freeze-thawed or heat-killed macrophages (but not freeze-thawed platelets or lymphocytes) were effective in restoration. Peripheral blood mononuclear leukocytes and splenic lymphoid cells were not active in the recipients syngeneic to the donor cells but were acitve in recipients allogeneic to the donor cells. Polymorphonuclear leukocytes isolated from peripheral blood or an inflammatory exudate were not active. Intact thymic function seems unimportant since antibody-mediated suppression took place as effectively in thymectomized mice as in normal controls.


The frequency of phosphorylcholine-specific B cells in conventional and germfree BALB/C mice. The frequency of B cells specific for phosphorylcholine (PC) in spleens of nonimmune conventionally-reared and germfree adult Balb/c mice was determined by an in vitro splenic focus technique. Analysis of spleen cells from 11 conventionally reared donors revealed an average of 19.1 PC-specific clonal precursor cells/10-6 donor B cells when calculated utilizing a cloning efficiency previously determined for the in vitro splenic focus technique of 4%. This number is one-tenth the frequency determined for 2, 4-dinitrophenyl- and 2,4,6-trintrophenyl-specific B cells; however, both idiotypic and isoelectric analysis reveal that the majority of B cells specific for PC yield monoclonal antibodies which share the TEPC 15 idiotype and contain a homogeneous IgM peak with a pK of 4.60. Thus this single clone appears on the average to be represented by more than 10-3 B cells in conventionally reared nonimmune adults. A similar frequency for precursor cells of this clonotype was found in an analysis of seven  germfree adult mice so that the high frequency of B cells of this colontype appears to be independent of previous antigenic stimulation. Among the 11 conventionally reared donors analyzed, frequencies varied from 1.6 to 52/10-6 B cells--an extreme degree of variation when compared to the almost invariant frequencies observed for 2, 4-dinitrophenyl, 2, 4, 6-trinitrophenyl, and fluoroscein. This variation is interpreted as a reflection of the life cycle of a single clone with a possible role for antigen implied by smaller variations observed in grem free mice.


Cellular cooperation during in vivo anti-hapten antibody responses. I. The effect of cell number on the response. Cellular interactions in adoptive secondary anti-hapten antibody responses to the hapten 2, 4-dinitrophenyl (DNP) have been studied. It was shown that DNP-specific B cells must interact with carrier specific helper T cells to give optimal responses. Independent titration of B cell and helper cell activity in adoptive anti-DNP antibody responses gave the following results: Doubling the number of transferred B cells approximately doubled the subsequent antibody response. Doubling the number of helper cells leads to nearly 4 times as much anti-DNP antibody, measured 7 days after boosting ("premium effect"). This marked effect of helper cell number on the antibody response is thought to be due primarly to the interaction of two populations of carrier-specific cells in the helper effect, or to the interaction of two activities of a single population of helper cells, namely clone activation and clone expansion. Only a very small proportion of the premium effect given by helper cells could be attributed to increases in antibody affinity.


Cellular cooperation during in vivo anti-hapten antibody responses. II. The effect of in vivo and in vitro x-irradiation on T and B cells. The effect of x-irradiation on T and B cells involved in an in vivo adoptive anti-hapten antibody response has been studied. In vitro x-irradiation inactivates both B cells and T cells. B cells are somewhat more sensitive than T cells to such treatment, the dose required to reduce B cell activity by 50% being about 50 R, whereas 200 R will reduce helper cell activity by 50%. By contrast, if recipient mice are primed with carrier, they develop helper activity that is totally resistant to 500 R x-irradiation within 3 days of immunization. This activity can not be transferred from irradiated, carrier-primed mice by either spleen cells or serum, although anti-carrier antibody is shown to give a modest helper effect late after boosting. However, this radioresistant helper effect can be reproduced in a cell transfer system by giving carrier-primed spleen cells to normal recipients that are then x-irradiated (500R) and given a large dose of carrier protein. Neither cells nor carrier alone will give rise to such radioresistant helper effects. Similar helper activity is occasionally observed when normal mice are irradiated 24 hr after being given helper cells, in the abscence of carrier and boosted shortly thereafter. The differences between these radioresistant helper effects, and the radiosensitivity of helper cells transferred after x-irradiation, may be related to x-ray induced changes in cell migration patterns.


Cellular cooperation during in vivo anti-hapten antibody responses. III. The helper cell activity of activated thymocytes, of spleen cells treated with anti-theta serum, and of spleen cells from anti-thymocyte serum-treated or adult thymectomized donors. An adoptive secondary anti-2,4-dinitrophenyl (DNP) antibody response involving T-B cell collaboration has been studied. In particular, attempts have been made to affect the unexpectedly steep log dose-response curve obtained when graded numbers of helper cells are transferred to irradiated recipients given a fixed number of B cells (premium effect). A variety of means were used to alter helper cell activity, and this activity was then measured quantitatively, as was the ability of the helper cells present after these treatments to give a premium effect. It was shown that activated T cells are approximately twice as active as spleen cells in helper activity and give a comparable premium effect. Graded doses of anti-theta serum plus complement markedly reduce the helper activity of spleen cells without affecting the premium effect given by the residual cells. Treatment of primed cell donors with limited doses of heterologous anti-mouse thymocyte serum (ATS) before transfer does not affect B cell activity, but readily inactivates helper cells, again without affecting the premium effect given by the residual cells. Adult thymectomy (ATx) of helper cell donors before priming with carrier led initially to increased helper activity relative to age-matched control donors. This increase may reflect the loss of nonspecific suppressor T cells from spleens shortly after ATx. Late after ATx, there was about a 2-fold decrease in helper activity, probably reflecting a loss of helper cell precursors. At no time was there any change in the premium effect. In view of the failure of any of the techniques used to abolish the premium effect given by helper cells in this response, it seems likely that this premium effect is due to the cooperative interaction of two very similar types of mature T cell. Alternatively, the premium effect observed here may result from the interaction of two activities of a single type of T cell which is mediated by independent factors.


Antibodies to thymocytes in sera of normal controls and of patients with schizophrenia. The sera from normal healthy individuals and from patients with schizophrenia were tested for cytotoxic antibodies against thymic antigens located on thymocytes. It was found that the sera from schizophrenics do not show higher cytotoxicity to mouse and rabbit thymocytes than the sera from normal controls. Furthermore, absorption of sera with adult mouse thymocytes and brain failed to show any difference between the sera from these two groups of subjects.


Ontogeny of mouse lymphocyte function. II. Development of the ability to produce antibody is modulated by T lymphocytes. The relative functional maturity of neonatal mouse spleen T- and B-cell populations was assessed by comparing the ability to respond to the thymic-independent antigen, DNP-Ficoll, or thymic-dependent SRBC by producing antibody in vitro. Although mouse spleen cells responded to DNP-Ficoll at an earlier age than they responded to SRBC or TNP-SRBC, the reason for the lag in the T-dependent response was confounded by the finding of high numbers of suppressor T lymphocytes in the neonatal spleen. Thus, small numbers of neonatal spleen T cells or thymocytes significantly decreased the in vitro antibody response of adult spleen cells. Although B lymphocytes appear to be functionally mature soon after birth, their acitivity may be modulated by an excess of suppressor T cells; e.g., the reconstitution of helper cell function in the neonatal spleen required anti-theta treatment before addition of adult helper cells. Suppressive activity attributable to T cells seems to play a dominant role in determining the ability of the neonatal animal to react positively or negatively to antigenic stimulation.


Expression of T-cell differentiation antigens on effector cells in cell-mediated cytotoxicity in vitro. Evidence for functional heterogeneity related to the surface phenotype of T cells. The cell-mediated cytotoxicity (CMC) of nonadherent cells from the peritoneal cavity (NAPC) of alloimmunized mice can be measured by the [3H]proline microassay. The exhibition of thymus-derived (T) cell antigens on these killer cells was studied by incubating them with the relevant T-cell antisera and complement (C), under optimal conditions for lysis, before performance of the CMC assay. Under these conditions, the following T-cell antigens were demonstrable on the killer population in terms of percent reduction in CMC by the respective antisera: (a) Thy-1.1 (83%) and Thy-1.2 (100%), (b) MSLA (86%), (c) NTA-RA (a T-cell antigen recognized by naturally occurring autoantibody of NZB mice) (62%), (d) Ly1.1 )58%, (e) Ly-2.1 (11%; considered a marginal result) and Ly-2.2 (63%), and (f) Ly-3.2 (77%). The following were not demonstrable: (g) TL, and (h) Ly-1.2. (i) The antigen Ly-3.1 was not studied. Omission of C deprived all T-cell antisera tested of their capacity to suppress CMC, indicating that the cell components recognized by such antisera may perform no direct function in CMC. On the assumption that all Ly+ cells are Thy-1+, it is clear that the T-cell members of the immune NAPC population must be heterogenous. This follows from the fact that the proportions of T cells lysed by different Ly antisera did not correspond with ensuing degree of loss of CMC capacity. The extremes were represented by anti-Ly-1.2 (74% Thy-1+ cells lysed, but no reduction in CMC) and Ly-3.2 (54% Thy-1+ cells lysed, with 77% reduction in CMC). From this initial survey it appears that the C57BL/6 mice killer T-cell population active in CMC in vitro is relatively rich in surface antigens of the  Ly-2/Ly-3 category and relatively poor in representation of the Ly-1 surface antigens. It remains to be seen whether this killer cell phenotype, poor in Ly-1 and rich in Ly-2/Ly-3, is characteristic of the mouse generally. From these results it appears that subsets of T cells with different immunological functions may exhibit qualitative or quantitative differences in surface antigens specified by different Ly loci; this will be easier to assess in the future when the results of experiments with the same Ly antisera but dealing with T-cell functions other than CMC become available.


Genetic regulation of the antibody response to H-2Db alloantigens in mice. I. Differences in activation of helper T cells in C57BL/10 and BALB/c congenic strains. B10.A(5R) mice immunized with C57BL/10 spleen cells demonstrate a normal T-cell-mediated cytotoxicity to H-2Db tumor cells but they do not mount any IgG antibody response to H-2Db alloantigens. B10.A(5R) mice do show a high titered IgG response when immunized with A.BY cells, which differ at H-2Db plus non-H-2 cell surface antigens, or with B10.A(2R) cells, which differ at H-2Db, H-2Kk, and H-2Ik cell surface antigens. These findings indicate a failure of the T-helper cells to induce the switch from IgM to IgG when the H-2Db alloantigens are the only difference on the immunizing cell. In immunizing H-2d mice with congenic H-g2 cells which differ only in the H-2Db region, mice of the C57BL/10 background made only IgM antibodies whereas mice of the BALB/c background made IgG antibodies. This comparison confirms that genes separate from H-2 regulate the T-cell helper function. The genes that influence the T-cell helper function do not regulate the T-cell-mediated cytotoxicity.


Rosette-forming ability of thymus-derived lymphocytes in cell-mediated immunity. I. Delayed hypersensitivity and in vitro cytotoxicity. Effector cells in delayed hypersensitivity and in vitro cytotoxicity were studied in lymph node cells from animals immunized with sheep erythrocytes (SRBC) in complete Freund's adjuvant. Delayed hypersensitivity response (DHR) was assayed by the increase in foot pad swelling after the intrafoot pad injection of immune cells plus antigen. Cell-mediated cytotoxicity against SRBC was assayed by a microcytotoxicity test with sheep fibroblasts as target cells. Effector cells were antigen specific, sensitive to anti-theta serum plus complement (C), and insensitive to anti-Ig serum plus C. A nonrosette-forming (non-RFC) small lymphocyte effector T cell and a rosette-forming medium lymphocyte effector T cell were isolated by velocity sedimentation. The small lymphocyte non-RFC required a longer time than the medium lymphocyte RFC effector cell to produce maximum activity. Buoyant density failed to distinguish medium lymphocyte effector cells in DHR and in vitro cytotoxicity.


Mechanical control of the time-course of contraction of the frog heart. Changes in load during most phases of an isotonic contraction of the frog and turtle heart increased or decreased the duration of the twitch. It was abbreviated by a maintained increase or by a brief decrease in load. The relaxing effect of these procedures developed with a delay lasting more than a second under some conditions and will be called lengthening deactivation. The reverse procedures, a maintained diminution or a brief increase in load, increased the duration of the twitch. This effect will be called shortening activation. Although the termination of relaxation may be delayed or advanced by the mechanical interventions mentioned, the normal time-course of isotonic relaxation was always resumed later, regardless of the starting level of the load, making it possible to measure accurately changes in the duration of the twitch. The responses to changes in load produce positive feedback during the isotonic contraction and explain, at least in part, the difference in the time-course of isotonic and isometric contraction. The effects of changes in load were much smaller and briefer in the atrium than the ventricle.


Absent vestibulo-ocular reflexes and acute supratentorial lesions. Loss of vestibulo-ocular reflexes occurred in two patients with acute supratentorial lesions who received therapeutic doses of anticonvulsant drugs. There was no clinical or angiographic evidence of focal brain-stem damage. Absence of vestibulo-ocular reflexes is attributed to a combination of acute cerebral damage and anticonvulsant drugs. The loss of these reflexes in patients with acute cerebral lesions cannot be interpreted as evidence of irreversible brain-stem injury.


Excitability modulation by taurine. Transmembrane measurements of neuromuscular transmission. The presence of taurine, a non-essential amino acid, in nerve and muscle has been previously associated with inhibition of activity in the central nervous system, with the etiology of epileptogenic foci, and with the muscle weakness of muscular dystrophy. We present here data showing a small and probably insignificant effect of taurine on neuromuscular transmission per se, but significant hyperpolarization of the membrane potential in both taurine-incubated and taurine-loaded muscles. In addition, we found that taurine reduces the time course of the muscle action potential. The results are interpreted in terms of neuromuscular transmission and excitation-contraction coupling consequent to these phenomena. This interpretation is compatable with the hypothesis that taurine is involved in the genesis of muscular dystrophy where the membrane potential is depolarized. Our results and interpretation can also explain the anti-arrhythmic action of taurine on cardiac muscle.