Effect of ampicillin and chloramphenicol alone and in combination on ampicillin-susceptible and -resistant Haemophilus influenzae type B. To evaluate ampicillin (Amp) and chloramphenicol (Cm) alone and in combination against Haemophilus influenzae type b, we examined the viability of 5 log(10) colony-forming units (CFU) of early-log-phase organisms per ml after 4 and 8 h of incubation with the drug(s). Nine Amp-susceptible (Amp(s)) and five Amp-resistant (Amp(r)) systemic isolates were examined. Antibiotic concentrations included: the minimum inhibitory concentration (MIC) of Amp, 50% of the MIC of Amp, 25% of the MIC of Amp, the MIC of Cm, 50% of the MIC of Cm, 25% of the MIC of Cm, and nine combinations of these concentrations. Both Amp and Cm at their MIC significantly reduced bacterial titers of Amp(s)H. influenzae type b after 8 h of incubation (1.36 and 1.47 log(10) CFU/ml, respectively; both p < 0.01); only Cm at its MIC significantly reduced the number of viable organisms after 4 h (0.91 log(10) CFU/ml; P < 0.001). With Amp(r) organisms, significant reductions in mean bacterial titers occurred after 4 and 8 h of incubation in the presence of Amp at its MIC (1.66 and 2.06 log(10) CFU/ml, respectively; both P < 0.02); smaller but significant reductions were noted after 4 and 8 with Cm at its MIC (0.56 and 0.87 log(20) CFU/ml, respectively; both P < 0.025). Antagonism with Amp(s) or Amp(r) strains was not seen. We conclude that combinations of Amp and Cm have indifferent effects on Amp(s) and Amp(r)H. influenzae type b.


In vitro activity and beta-lactamase stability of cefazaflur compared with those of beta-lactamase-stable cephalosporins. The in vitro activity of cefazaflur, a parenteral cephalosporin, was determined against 590 clinical isolates. Cefazaflur inhibited the majority of gram-positive cocci at concentrations below 1 mug/ml except for enterococci. The agent was as active as cefamandole or cefoxitin against most Escherichia coli, Klebsiella pneumoniae, and Proteus mirabilis. Although it inhibited a number of strains of Enterobacter, indole-positive Proteus, and Serratia resistant to cephalothin, it was much less active against these organisms than were cefamandole or cefoxitin.


Homobodies: do they exist?

Homobodies: do they exist? Within an immunological network consisting of interacting paratopes and idiotopes, three families of molecules can be postulated which bear a peculiarly close connection to any given epitope; the antibody, which carries paratopes which fit the epitope; the anti-idiotype, which carries paratopes fitting the idiotopes of the antibody; and the "homobody", which carries idiotopes to which the paratopes of the antibody fit. The homobody, called by Jerne the "internal image set", is a molecule which resembles the epitope, not like a mould resembles its cast, but like one cast resembles another cast made in the same mould. A distinction between homobody and anti-idiotype poses certain problems which are discussed in the light of a hypothetical experiment using avidin as the antigen.


TNS-evoked long loop effects.

TNS-evoked long loop effects. Local analgesia can be produced by transcutaneous electrical stimulation of peripheral nerves. This is used in the treatment of chronic pain states. Its clinical effectiveness depends on two points; namely (1) the stimulation has to be perceptible, and (2) paresthesias elicited by TNS must be localized in the area of pain. To verify this in healthy subjects we produced an experimental pain by radiant heating of the skin and tested the analgesic effect of TNS. TNS stimuli parameters (duration, amplitude and frequency) were determined so that double blind conditions were given. Stimulation with small rectangular pulses showed the best analgesic effect especially at a stimulation rate of 100 Hz. The stimulation of various nerves showed that most of the analgesic effects depend on spinal level mechanisms but probably long loop effects are involved.


Studies of calcitonin cells and parathyroid glands of the Indian palm squirrel, Funambulus pennanti in response to experimental hypercalcaemia. Long term hypercalcaemia was induced in F. pennanti by alternate day intramuscular injections of 50,000 IU of vitamin D2 and by giving them 1% CaCl2 solution prepared in tap water to drink. The controls were not injected with vitamin D2 and were given tap water. The serum calcium levels at various stages of the experiment (1-29 days) show increased values as compared with those of control animals. The calcitonin cells in the treated animals generally exhibit an increase in their number up to the 15th day. Mitotic figures are also encountered between the 7th and the 15th day of treatment. This exhibits the increase in the number of C cells. Constant calcium challenge results in increased quantities of secretory granules among these cells up to the 15th day and in degranulation from the 17th day onwards. It also causes degenerative changes in a certain number of C cells. The parathyroids exhibit atrophic changes (25 days onwards) due to chronic hypercalcaemia. For short term hypercalcaemia, animals were injected intravenously with 1 ml of 10% solution of calcium gluconate. The calcitonin cells do not exhibit any change during the first half hour but thereafter they exhibit progressive degranulation, resulting in marked degranulation after 5 hours of the injection. The parathyroids remain unaffected throughout the experiment and show no histological change.


Anastomotic ulceration following subtotal and total pancreatectomy. Ulcerogenic potential exists after subtotal gastrectomy, pancreatic resection and separation of the pancreatic and bile ducts from the gastric outlet. Collectively, as with radical pancreatoduodenectomy or total pancreatectomy, a substantial risk of an anastomotic ulcer developing can be expected. To elucidate this, we reviewed the records of 297 patients who had undergone either radical pancreatoduodenectomy (Whipple procedure) or total pancreatectomy. We found that 18 patients (6%) had developed an anastomotic ulcer and that total pancreatectomy was statistically more ulcerogenic than a Whipple procedure. An anastomotic ulcer a virulent complication; two-thirds of the patients had bleeding, and it was a contributing cause of death in 22% of patients in whom ulcer developed. In patients who have a favorable prognosis, the addition of vagectomy to either procedure should be considered.


Protease inhibitors and experimental acute hemorrhagic pancreatitis. Proteolytic enzyme inhibitors have been reported to decrease morbidity and mortality from certain types of experimental pancreatitis, although recent randomized trials have been unable to demonstrate that they are of benefit in the treatment of clinical acute pancreatitis. We have evaluated the effect of two proteolytic enzyme inhibitors (trasylol and chlorophyll-a, on experimental acute pancreatitis induced in mice by the feeding of a choline-deficient ethionine-enriched diet. The mortality rate and the biochemical and morphological severity of pancreatitis were not altered by either trasylol or chlorophyll-a administration. Thus, in this respect, diet-induced pancreatitis appears to resemble clinical acute pancreatitis. The reasons for the lack of effectiveness of proteolytic enzyme inhibitors in the treatment of both forms of pancreatitis are discussed.


Use of Doppler ultrasound in intraoperative localization of intestinal arteriovenous malformation. Precise localization of an intestinal arteriovenous malformation is essential for the complete resection of the lesion. Identification at operation can be difficult, and intraoperative angiography is not without complications. Blood flow within the small vessels of the bowel wall can be easily detected by Doppler ultrasonography, and arterial flow can be distinguished from venous flow. Intraoperative use of the Doppler ultrasound technique localized an AVM in the small intestine. Distinctive flow signals were present over the intestinal AVM which allowed it to be easily differentiated from adjacent normal intestine. Confirmation was secured by measuring the PO2 of blood aspirated from the vein draining the involved segment and comparing it to the PO2 from a vein draining an adjacent segment.


Role of arteriography in rectal hemorrhage due to arteriovenous malformations and diverticulosis. Nine patients with arteriographic right colon arteriovenous malformations (AVM's) are reviewed. Even though five of the eight surviving patients had diverticulosis in remaining colon, right colectomy with removal of the AVM effectively controlled bleeding in seven of the eight patients in follow-up intervals of 14-66 months. Such results suggest that right colon AVM, rather than diverticulosis, may be the more common cause of chronic recurrent lower intestinal hemorrhage in the elderly. Rebleeding occurred in one patient and has heretofore been reported in seven cases. However, since AVM bleeding is rarely life-threatening, right colectomy for chronic recurrent rectal bleeding is justifiable as the initial operative procedure for an AVM, even in the presence of left colon or sigmoid diverticulosis. In the same five year period, 1973-1977, four additional patients with massive hemorrhage, characteristic of diverticulosis, had emergency arteriography with demonstration of a localized site of diverticular bleeding that was controllable by partial colon resection. Consequently, segmental resection of the colon, utilizing selective arteriography, has thus far eliminated bleeding in eight of nine patients with diverticulosis. The judicious use of arteriography in patients with diverticulosis and rectal bleeding substantially reduces the requirement for subtotal colectomy and ileoproctostomy.


The role of cardiovascular hemodynamics and liver histology in evaluating bleeding cirrhotic patients. Preoperative cardiovascular hemodynamics and percutaneous liver biopsies were used to evaluate the pathophysiologic factors determining the operative prognosis of patients with cirrhotic liver disease and bleeding esophageal varices. These studies confirm the observations of Siegel that the greater the magnitude of the peripheral abnormalities in vascular tone and oxygen consumption the better must be the capability of the ventricular function, if the cirrhotic is to survive emergency or urgent portal decompressive surgery. These studies also show that the cardiovascular hemodynamics are directly correllated with the nature and degree of the abnormalities in the liver biopsy, and that pathologic and physiologic features of this disease which impact on surgical prognosis can be expressed through the easily obtained Survival Index. Bleeding cirrhotic patients with poor quality hemodynamics and poor histologic characteristics should be treated non operatively, since the operative mortality appears greater than that produced by a strategy of medical supportive therapy and delayed surgery if stabilization occurs.


Saphenous vein bypass from the aorta to the anterior interventricular vein. A patient is described who had myocardial revascularization with saphenous vein grafts. One of the grafts was inadvertently anastomosed to the anterior interventricular vein instead of the anterior descending coronary artery. He also underwent saphenous vein bypass to the right coronary artery and resection of a left ventricular aneurysm. The patient remained free of angina up to 3 1/2 years after operation, at which time he was seen with tachyarrhythmia and a continuous murmur. Coronary arteriography revealed patent bypass grafts to both the distal right coronary artery and the anterior interventricular vein. Whether the patient benefited from the graft to the anterior interventricular vein remains indeterminable because of the successful bypass to the right coronary artery and the left ventricular aneurysmectomy. He did experience clinical improvement for 3 1/2 years, and the symptom when he was seen at our institution was tachyarrhythmia, not chest pain. Additionally, his exercise tolerance had improved markedly. We believe the potential advantages of bypass from the aorta to the coronary veins is still unexplored.


Effects of cardioplegic solution on human contractile element velocity. A technique for measuring the maximum contractile element velocity (Vpm) of the myocardium was developed, verified, and employed in patients to allow accurate intraoperative assessment of the adequacy of myocardial protection. Four groups of patients were studied. Ten patients had coronary artery bypass grafts (CABG) with cardioplegia; 13 had CABG with coronary perfusion, ventricular fibrillation at 28 degrees C, and aortic clamping for distal anastamoses; 6 had aortic valve replacement (AVR) with cardioplegia; and 7 had AVR with coronary perfusion to the beating heart. For cardioplegia, a solution of 5% dextrose in 0.2% saline at 4 degrees C with 25 mEq of potassium chloride and 12.5 gm of mannitol was infused initially, followed by 500 ml every 30 minutes. Clinically all patients did well, and there were no deaths. Patients having CABG with intermittent coronary perfusion during ventricular fibrillation had significant (p less than 0.01) depression of Vpm from 38.3 to 30.8 sec-1 while Vpm in patients having CABG with cardioplegia was unchanged. Patients having AVR with continuous coronary perfusion or with cardioplegia (average anoxia time, 70.4 minutes) had no significant change in Vpm. We conclude that this cardioplegic solution provided adequate protection of myocardial function for up to 105 minutes of continuous aortic clamping in humans. The depression in Vpm observed following CABG with intermittent coronary perfusion is consistent with previous suggestions that this combination is detrimental because of maldistribution of coronary blood flow during ventricular fibrillation.


Reduction of intraoperative myocardial infarction by means of exogenous anaerobic substrate enhancement: prospective randomized study. In a prospective study, patients who had an ejection fraction of 40% or more and who were undergoing elective coronary artery operation were randomly divided into three groups that differed in the method of anaerobic substrate enhancement during cardiopulmonary bypass. Group 1, the controls (n = 157), received no additional glucose, insulin, and potassium solutions and experienced immediate spontaneous defibrillation (10%), transmural myocardial infarction (10.3%), malignant ventricular arrhythmias (26%), and severe atrial arrhythmias (20%). Group 2 (n = 120) received a bolus of hypertonic glucose, insulin, and potassium in the pump perfusate before aortic cross-clamping. In this group, the rate of spontaneous defibrillation was 41%, of transmural infarction, 8.3%, of malignant ventricular arrhythmias, 31%, and of severe atrial arrhythmias, 19%. Group 3 (n = 114) had the aortic root continuously infused with glucose, insulin, and potassium solution at 4 degrees C during aortic cross-clamping. This group was significantly improved; the rate of spontaneous defibrillation was 60%, there were no transmural myocardial infarctions and the incidence of severe atrial arrhythmias was 6% and that of malignant ventricular arrhythmias, 5%. It is proposed that the superior clinical results in Group 3 resulted from better myocardial preservation achieved by more efficient means of providing continuous anaerobic substrate, coronary washout, and elution of lactic acidosis, uniform global hypothermia, and direct supplemental myocardial potassium in addition to mere cardioplegic effects.


Ventricular venting during coronary revascularization: assessment of benefit by intraoperative ventricular function curves. Although the physiological benefits of left ventricular venting during cardiopulmonary bypass have been documented experimentally, air embolus is still a concern clinically, and surgeons, therefore, continue to debate whether or not to vent routinely during coronary revascularization. In this study, 10 patients vented during revascularization and 10 not vented were studied immediately before and after cardiopulmonary bypass using Sarnoff ventricular function curves to assess changes in ventricular performance. Stroke work index by each patient before and after bypass was compared at common left atrial pressures and concomitant similar systemic resistances. In patients who were vented, there was overall no change in function (98 +/- 7% of control; range, 146--73%). In patients who were not vented, there was overall significant depression of function (67 +/- 5% of control; range, 91--45%, p less than 0.01). The overall absence of depression in patients who were vented, several of whom had very poor ventricles before bypass, strongly supports venting for coronary revascularization.


Acute effects of aortocoronary bypass surgery on left ventricular function and regional myocardial mechanics: a clinical study. The acute effects of myocardial revascularization on overall left ventricular performance and on myocardial segmental wall motion were assessed intraoperatively in 22 patients who had unstable (11 patients) or stable angina pectoris (11 patients). Segmental contraction patterns were evaluated using an ultrasonic transit-time method. In 9 patients with unstable angina pectoris, notable improvement in segmental wall motion was observed as the short-term response to coronary bypass grafting. Hypokinetic patterns were rendered normal after revascularization. Despite marked changes in segmental myocardial function, overall left ventricular performance was not altered notably. In contrast, reperfusion did not lead to acute effects on either segmental wall motion or total left ventricular function in patients with stable angina pectoris. The results indicate that aortocoronary bypass grafting may improve segmental wall motion in patients with unstable angina.


Early pleural effusion in histiocytosis X. Two cases of histiocytosis X, one of the disseminated form and the other localized to the lung, are reported, in which pleural effusion, probably due to pleural involvement as assessed by biopsy, was one of the initial manifestations of the disease.


Structural abnormalities in the cerebral cortex of chronic schizophrenic patients. Enlarged cerebral ventricles in chronic schizophrenic patients suggest a process of mild cerebral atrophy occurs in some. To see if this process involves the cerebral cortex, the widths of the Sylvian fissure, the interhemispheric fissure, and three cortical sulci were measured blindly on computerized tomography (CT) scans of 75 chronic psychiatric patients and 62 asymptomatic volunteers, all less than 50 years of age. A total of 19 of the 60 patients with chronic schizophrenia had at least one abnormality. All 15 patients with other diagnoses were within the control range. Comparing those chronic schizophrenic patients with abnormalities to those without them, there were no significant differences in age, length of illness or treatment, and length of hospitalization. From this and ventricular size data, two thirds of the chronic schizophrenics had some cerebral structural abnormality. Ventricular enlargement did not correlate significantly with cortical abnormalities. Therefore, more than one etiology may account for the structural abnormalities found in chronic schizophrenic patients.


Immunological studies in hairy cell leukemia. Cytochemical and immunological studies were performed on tissues and mononuclear cell suspensions from ten patients with hairy cell leukemia. In all cases studied, tartrate-resistant acid phosphatase was noted within the cytoplasm of hairy cells (HCs). In two thirds of the cases, alpha naphthyl acetate esterase was observed in HCs. In addition, HCs did not form spontaneous rosettes with sheep erythrocytes. A variable number of HCs displayed complement receptors. The nonspecific binding of conjugated immunoglobulin to HCs probably reflected the presence of a high concentration of Fc receptors on the HCs surface. In three cases, the conjugated immunoglobulin reacted predominantly to one light chain, thus suggesting the presence of a monoclonal immunoglobulin. In four of six cases, mononuclear cell suspensions of HCs demonstrated latex phagocytosis. In one case, HCs displayed resynthesis of surface membrane immunoglobulin, the presence of B cell antigen, and phagocytosis of latex. These findings suggest that HCs are distinctive and contain properties of both B lymphocytes and monocytes.


Coincidental hairy cell leukemia and large cell malignant lymphoma. A 64-year-old man who had a two year history of hairy cell leukemia became feverish with night sweats. Splenomegaly and enlarged nodes in the neck and in the anterior mediastinum were detected. Splenectomy was performed and multiple white nodules on a dark-red background were seen in the spleen. The white nodules represented a large cell malignant lymphoma; other areas of the spleen contained mononuclear cells typical in hairy cell leukemia. One of the hilar lymph nodes of the spleen was involved by malignant lymphoma, and the other eight lymph nodes were involved by hairy cell leukemia. The coincidence of the two diseases tends to favor the view that hairy cells are of lymphocytic origin.


[Course of metastatic osteogenic sarcoma treated with chemotherapy only (author's transl)]. The case of a six year old girl with metastatic (or primarily multifocal) osteogenic sarcoma is reported, which was treated with chemotherapy only. Within 15 months--the primary tumor regressed and at least a transient inactivation has resulted until now;--the pulmonary metastases showed no progression;--an osteoplastic destruction of the sacrum has completely regressed. At the time of report (15 months after initiation of treatment) neither on X-ray controls nor on bone scan signs of tumor regrowth are evident. The child is without any complaints and the leg has no loss of function.


Portal vein thrombosis associated with cirrhosis: clinical importance. A group of 86 cirrhotics undergoing therapeutic variceal decompressive procedures were studied. Patients with portal vein thrombosis (PVT) comprised 21% of the group and more frequently had uncontrollable hemorrhage at an earlier stage of liver disease. Mortality in the 18 cirrhotics with PVT was higher (56%), mostly as a result of rebleeding. Despite adjustments for stage of liver disease and type of operation, in no cirrhotic with PVT did postshunt encephalopathy develop, compared with 32% incidence in patients without PVT (P less than .05). It is concluded that patients with cirrhosis and PVT represent a different subpopulation of cirrhotics. Once adequate variceal decompression has been achieved, their prognosis should be superior to cirrhotics without PVT because their hepatic hemodynamics are unaffected by total shunting, hence precluding further impairment of liver function as a result of acute reduction of hepatic blood flow.


Aortocystoduodenal fistula: rare complication of pancreatic pseudocyst. An upper gastrointestinal tract hemorrhage resulting from an aortocystoduodenal fistula developed in a patient with a pancreatic pseudocyst. The fistula was exposed through a duodenotomy, necrotic material was debrided from the pseudocyst and the aortic wall, the aortic defect was closed primarily, and the pseudocyst was drained through a cystoduodenostomy. The case is discussed as a rare hemorrhagic complication of pancreatic pseudocysts and as an illustration that under certain circumstances of infection in areas where aortic bypass can be difficult to perform, primary vascular repair can be a successful method of managing aortoenteric fistulas.


Drug actions in rats with arteriosclerosis induced by toxic doses of vitamin D2. The vascular response of isolated perfused hind legs from normal and arteriosclerotic rats to noradrenaline, ATP, PGF2 alpha and vasopressin was determined. The increase of perfusion pressure to noradrenaline and ATP was reduced and that to PGF2 alpha and vasopressin was enhanced in arteriosclerotic rats in comparison to normal animals. The results indicate that the reactivity of the vascular smooth muscle of isolated hind legs of arterioslerotic rats is not only quantitatively but also qualitatively different in comparison with normal rats.


[Blood vessels and nerve apparatus of the heart and small intestine during continuous experimental bypass shunting]. In 140 animals (mongrel dogs of different age and sex) by means of histological, neurohistological, histochemical, histoenzymatic and electron microscopic methods, character and dynamics in changes of nerve elements and blood vessels of the heart and the small intestine have been studied under the conditions of revascularization at aorto-coronary and aorto-mesenteric shunting. The changes revealed are mainly reactive in their character, perform defensive, compensatory-adaptive response of the organs to the orative trauma and to an adequate but slightly changed blood stream. The degree of acuteness and the time when these changes disappear depends on duration of postocclusive ischemia, on the organs' resistivity to its effect on the operative trauma. Operation of aorto-coronary and aorto-mesenteric shunting are rather effective and protect the vascular bed and the intramural nerve appratuses of the heart and the small intestine from severe destructive disorders developing as a result of occlusion in the arterial vessels.


The Campomelic syndrome. Temporal bone histopathologic features and otolaryngologic manifestations. The campomelic syndrome is characterized by dwarfism, craniofacial anomalies, bowing of the tibiae and femora, cutaneous dimpling overlying the tibial bend, respiratory distress, and early death. Otolaryngologic manifestations include flat facies with a broad nasal bridge, low-set ears, cleft palate, mandibular hypoplasia, and tracheobronchial malacia. The underlying pathologic feature appears to be disturbance in cartilage growth involving the affected bones and the respiratory tract cartilage. The cause is unknown. We report clinical and histopathologic features in two cases of this syndrome. The endochondral layer of the otic capsule contained no cartilage cells. The cochlea was short and flattened, presenting a scala communis. The vestibule and the canals were deformed by bone invasion. Defective endochondral ossification of the petrooccipital synchondroses possibly explains the shortened skull base seen in this syndrome. The tracheobronchial malacia significantly contributes to respiratory distress and neonatal death.


The relevance of 25-hydroxycalciferol measurements in sera of patients with renal failure. Iliac crest biopsies and serum specimens were obtained from 36 non-dialyzed uraemic patients. The mean serum 25-hydroxycalciferol concentration of the patients was lower than that of normal subjects and a significant correlation was found between 25-hydroxycalciferol values and the severity of osteomalacia. Parathyroid osteopathy was significantly correlated with serum immunoreactive parathyroid hormone but not with serum 25-hydroxycalciferol values. While normal serum 25-hydroxycalciferol concentrations in uraemic patients do not exclude the presence of osteomalacia, a low concentration is virtually diagnostic of this disorder.


The Mallory-Weiss lesion as a cause of upper gastrointestinal bleeding. A prospective study of patients with upper gastrointestinal bleeding admitted to a haematemesis and melaena unit has revealed an incidence of Mallory-Weiss tears of 8% (59 of 762 patients undergoing endoscopy). Prior vomiting was present in 60% and an associated upper gastrointestinal lesion in 44 percent. The majority of patients had a recent ingestion of alcohol and/or analgesics, whilst 34% had chronic heavy alcohol intake. Approximately 50% of patients required no blood transfusion, while 37% had over three units of blood. No patient in the group required surgical intervention, and one patient died because of general debility. This study suggests that the Mallory-Weiss tear accounts for a significant proportion of patients admitted with upper gastrointestinal bleeding, but that the mortality and morbidity are low.


Inferential therapy to promote union of mandibular fractures. Non-union of mandibular fractures is uncommon, but when it does occur it requires protracted treatment including further surgery. Nine patients with factors known to predispose to non-union out of 150 consecutive mandibular fractures received interferential therapy (I.T.) during the fixation period; all fractures united satisfactorily. In a retrospective study of 150 consecutive mandibular fractures previously treated by the same surgeons without I.T., three fractures resulted in non-union requiring grafting. Thus the incidence of non-union was 0% when I.T. was used and 2% in the control group.


Myocardial infarct imaging after cardiac surgery. One hundred and nineteen patients undergoing cardiac surgery had postoperative myocardial imaging performed with technetium pyrophosphate in order to assess the incidence of perioperative myocardial infarction. Fifty-six patients had only coronary artery bypass graft (CABG) surgery, of whom 13(23%) had a positive scintigram. Thirteen patients had CABG with other cardiac surgery and six (46%) had a positive scintigram. Fifty patients had other cardiac surgery but no CABG, and of these eight (16%) had a positive scintigram. The overall incidence of positive scintigrams was 23%, whereas definite or probable ECG diagnosis of infarction was present in 14 patients (12%). Serum levels of cardiac enzymes were higher in patients with positive scintigrams, but this finding did not consistently reach statistical significance. The use of a left ventricular vent during surgery did not correlate with a positive scintigram, nor did the total time on cardiopulmonary bypass or aortic cross-clamping. Patients having cardiac surgery, including CABG and valve replacement, have a 23% overall incidence of positive scintigrams. This suggests that the incidence of infarction after cardiac surgery is higher than can be recognized from the conventional criteria of ECG and enzyme changes.


Aortic valve replacement in a patient with von Willebrand's disease. Few cases have been reported to date of cardiac surgery performed on patients with von Willebrand's disease. This report details the management of such a patient, who underwent aortic valve replacement for severe calcific aortic stenosis.


Platelet aggregation and aggregation inhibition by different antiglobulins and antiglobulin complexes from sera of patients with rheumatoid arthritis. Antiglobulin complexes were isolated from rheumatoid arthritis (RA) sera in order to investigate their ability to aggregate human platelets and to influence platelet aggregation induced by heat-aggregated IgG. Eleven of 38 seropositive RA sera showed a significant platelet aggregation (PA) titer as compared to 40 normal control sera. No correlation was observed between sheep T cell agglutination titers and PA titers of individual sera. Ten of 22 RA antiglobulin preparations containing IgG, IgM, and IgA antiglobulins and antiglobulin complexes also showed positive PA tests. Such preparations were able to inhibit PA brought about by heat-aggregated human IgG. When IgG and IgM antiglobulins were tested separately, only IgM antiglobulins showed this inhibitory effect, whereas IgG antiglobulins were inactive. Neither IgG nor IgM antiglobulins induced PA alone. Reassociated IgG antiglobulin complexes consisting of carefully prepared IgG and isolated IgG antiglobulins were able to induce PA. Platelet aggregation by RA sera was thus shown to be due to IgG antiglobulin complexes present in the sera. It can be concluded from these experiments that the composition of antiglobulin complexes in individual sera comprised of antiglobulins of different classes and IgG "antigen" is responsible for PA results in vitro. In pathophysiologic terms, the reaction of IgG antiglobulin complexes with thrombocytes in vivo may be an important step in the chain of events leading to generalized vascular damage and deposition of immune complexes in vessel walls which is thought to be responsible for the development of generalized vasculitis in RA.


A review of the use, evaluations, and criticisms of the preliminary criteria for the classification of systemic lupus erythematosus. The use of the Preliminary Criteria for the Classification of Systemic Lupus Erythematosus (SLE) has been reviewed. Most authors describing 10 or more patients have used the criteria. Although originally designed for classification of patients, the criteria have often been used for the diagnosis of SLE. The sensitivity of the criteria was evaluated and found to be similar to the 90% reported in the original ARA trial. Studies on specificity yielded conflicting results. Various suggestions to improve the criteria have been made, including a more detailed description of some of the organ involvements and the adoption of laboratory tests that have been improved since or were unavailable at the time of original publication of the criteria.


[Simultaneous improvement of mood and release of growth hormone by L-5-hydroxytryptophan (Ro 3-5940) in normal subjects (author's transl)]. Infusion of a new soluble ester of L-5-hydroxytryptophan produced euphoria in 34/35 experiments in healthy subjects. Parallel measurements of growth hormone and mood changes showed a similar rise and fall of these two parameters in 8/11 subjects. These results indicate that central stimulatory serotoninergic mechanisms (in addition to the well-known dopaminergic and alpha-adrenergic stimulation) play a role in the control of growth hormone release.


Biophotometric investigations on the influence of cephradine and related cephalosporins on the growth of beta-lactamase-producing staphylococi. Biophotometric growth studies were conducted with clinical isolates of beta-lactamase-producing staphylococi in the presence and absence of cephradine, cephalexin, cefazolin and cephacetrile. Of the four antibiotics, cephradine was the most effective in prolonging the suppression of bacterial growth resulting from antibiotic action while cefazolin was the least effective. Cephalexin and cephacetrile were intermediate in effect. The superiority of cephradine over cephalexin was shown to be related to the more rapid destruction of cephalexin by beta-lactamase. On comparing the results of these studies with clinical experience it was concluded that biophotometric studies have clinical relevance and consequently should be used for evaluating the efficacy of beta-lactam antibiotics against pathogenic staphylococi.


Recognition and quantification of myocardial injury by means of plasma enzyme and isoenzyme activities after cardiac surgery. Serial plasma enzyme determinations were carried out in 32 patients who underwent cardiac surgery with the aid of extracorporeal circulation. Plasma creatine kinase (CK), the cardiospecific isoenzyme of CK (CKMB), and alpha-hydroxybutyrate dehydrogenase (HBDH) were determined from the onset of surgery up to 100 to 120 hours after operation. From the plasma enzyme activities, the total amount of enzyme released by the injured heart into the circulation could be calculated using mathematical equations solved numerically by means of a computer. The calculated amount of CK, CKMB, and HBDH released by the heart correlated well with (1) postoperative mortality, and (2) peak activities of the respective enzymes. The calculated amount of any of the 3 enzymes released showed poor or no correlation with (1) electrocardiographic criteria of myocardial infarction, (2) duration of cardiopulmonary bypass, and (3) duration of total aortic cross-clamping. This study shows that the extent of myocardial injury after surgery can be assessed quantitatively using the calculated amounts of enzyme released, as well as using peak plasma activities of CKMB and HBDH.


[Pleural empyemas in children due to Hemophilus influenzae]. 22 consecutive cases of pleural empyema due to H. influenzae in children are reported. An increment of its incidence during the 1975-76 period is observed. All cases were in children under 3 years of age, with a mean of 15.5 months. The presenting syndrome at admission was varied. In 50% of cases, pleuropulmonary infection was ignored. Nearly one half of cases of pleural empyema due to H. influenzae had simultaneous purulent meningitis caused by the same microorganism. This type of empyemas, though having a prolonged evolution, apparently appear to have a good prognosis, compared with that produced by S. aureus. There are some differences, being the most outstanding: low frequency of pyoneumothorax and the lack of radiological evidences of abscesses and or pneumotoceles, in any phase of the clinical course. Stress is placed on the value of the bacteriological study of blood and CSF in children under 3 years of age with pleural empyema. A high rate of positive blood cultures was found. (75%). Pieuropulmonary complications, as a frequent event during severe infections (septicemic disease) due to H. influenzae is considered. A clinical characterization of children in whom a pleural empyema could occur is proposed. A discussion is made about diagnostic, therapeutic and prognostic implications of these complications.


Release of soluble "blocking" and "suppressor" factors from normal lymphocytes treated with RNA from spleens of tumour-bearing mice. RNA extracted from the spleens of tumour-bearing (TLRNA) and tumour-immune (ILRNA) mice was shown to transfer to normal lymphocytes (NL) the ability to produce factors that blocked specific tumour-cell cytotoxicity and mediated specific antibody-dependent cell cytotoxicity (ADCC). Aliquots of normal C3H mouse lymphocytes were treated with TLRNA or ILRNA and cultured in vitro in the absence of tumour antigen. Supernatants were collected at 24h intervals and tested in a microcytotoxicity assay for blocking and ADCC activities. Factors that inhibited tumour destruction by specifically sensitized lymphocytes at the level of both the tumour cells and effector cells were demonstrable in culture supernatants of NL pretreated with TLRNA (50 or 100 microgram/4 X 10(6) cells) but not ILRNA. However, treatment of NL with either RNA resulted in the production factors that mediated tumour-specific ADCC. Cytotoxicity testing and absorption studies of the tumour cell and a control cell (LM) indicated that factors mediating ADCC and blocking at the target-cell level were specific for the tumour. Suppressor activity at the effector-cell level was not absorbed by tumour cells and represents a separate and distinct mechanism of immunosuppression. These data indicate that RNA faithfully transfers "suppressive" as well as "positive" types of immune responses that have been reported previously for lymphocytes obtained directly from tumour-bearing and tumour-immune animals.


Hairy cell leukaemia: surface markers and functional capacities of the leukaemic cells analysed in eight patients. In eight patients with hairy cell leukaemia (HCL) peripheral blood cells and in two patients also spleen cells were analysed for surface markers and functional capacities. Only cells containing the tartrate resistant isoenzyme 5 of the acid phosphatase were considered. Hairy cells (HC) of all patients were found to adhere spontaneously to glass and plastic surfaces and to spread after adherence like monocytes. They ingested latex particles of more than 1 micron diameter, but, in contrast to monocytes, did not phagocytose erythrocytes sensitized either by IgM or by IgG antibodies. HC of all patients bore Fc-receptors with a high binding affinity for aggregated IgG. Using 125 I-labelled F(ab')2-fragments of monospecific antibodies in autoradiography, only one light chain type was detected on HC of individual patients. In four patients mu- and delta-chains were simultaneously expressed on HC, whereas in two patients only gamma-chains and in one case only mu-chains were observed on HC. One patient showed a combination of gamma- and delta-chains on his HC. A great variation in density of surface immunoglobulings of HC was observed within individual patients. After removal by capping, surface immunoglobulin reappeared on HC during cell culture, but more slowly than on normal B-lymphocytes. As shown in two patients by internal labelling, HC secreted immunoglobulin light chains, but no heavy chains. On the basis of these findings the classification of HC as belonging to the B-cell lineage, rather than to the monocytic lineage, seems to be justified.


An immunoradiometric assay for procoagulant factor VIII antigen: results in haemophilia, von Willebrand's disease and fetal plasma and serum. An immunoradiometric assay (IRMA) has been developed based on the inhibitor which arose in a polytransfused severe haemophiliac. The two-site IRMA measures antigens closely associated with the procoagulant parts of the factor VIII complex, which are termed FVIIC antigens or FVIIICAG. FVIIICAG was present in normal plasma and also, at a slightly lower concentration, in normal serum. In 37 patients with haemophilia A, 36 had FVIIICAG levels of less than 10% of the normal plasma pool. In patients with von Willebrand's disease the levels of FVIIIC and FVIIICAG were in good agreement, both before and after treatment with cryoprecipitate or DDAVP. FVIIICAG was relatively stable in plasma at 37 degrees C and could also be detected in cord and fetal serum. The assay is of potential value for detecting reduced levels of factor VIII, for carrier detection and for the prenatal diagnosis of haemophilia.


Anterior membrane dystrophy following cataract extraction. This paper describes a patient with previously normal corneas who underwent bilateral cataract surgery. This was followed by acute dehiscence of the epithelium and the eventual appearance of fingerprint lines, microcysts, and map-like areas typical of Cogan's microcystic dystrophy. Light and electron microscopy showed the presence of epithelial microcysts and a seam of subepithelial material like basement membrane. It is suggested that this patient represents a rapid transition from apparent normality to an extensive anterior membrane dystrophy, precipitated by operation for cataract.


Punctiform and polychromatic pre-Descemet's dominant corneal dystrophy. A new type of pre-Descemet's corneal dystrophy is described. The opacities are punctiform, polychromatic, of uniform size, and evenly distributed over the whole cornea. The diagnosis is made only by slit lamp because there is no visual impairment. The disease is hereditary and follows the autosomal mode of inheritance with a high percentage of penetrance, expressivity, and specificity in 4 successive generations, in which 8 affected members were observed among a total of 46.


L-Phenylalanine ammonia-lyase and pisatin induction by 5-bromodeoxyuridine in Pisum sativum. The substitution of the base analogue 5-bromodeoxyuridine (BrdU) for thymidine in the DNA of pea pods (Pisum sativum) induces or enhances the level of phenylalanine ammonia-lyase (PAL) and also induces the phytoalexin, pisatin, a product of the same metabolic pathway. Cordycepin, a polyadenylate inhibitor at the RNA level, is a potent inhibitor of pisatin synthesis. Kinetic studies on the inhibition of the PAL-pisatin production by hydroxyurea indicate that BrdU must be incorporated into DNA before any induction takes place. 5-Iododeoxyuridine is also an inducer while 5-fluorodeoxyuridine is ineffective when applied alone. BrdU is incorporated into the DNA of pea cells and the nuclei undergoes condensation just prior to the detection of the induced responses.


Post-transcriptional control of messenger RNA diversity in frog embryos. The control of mRNA diversity during frog development has been investigated. Nuclear and messenger RNA from the early neurula and larval stages of Rana pipiens were hybridized in vast excess to labeled single-copy DNA and resistance to S1 nuclease was measured. Mixtures of RNA populations were also hybridized with single-copy DNA as a measurement of sequence overlap. Neurula and larval nuclear RNA hybridize to 11.3% and 12.1% of the single copy DNA. A mixture of both nuclear RNA populations hybridizes to 10.8% of the DNA, indicating a great amount of sequence overlap between the two populations. The mRNA complexity almost doubles during this developmental period from 4.7% of the single-copy DNA complexity at the early neurula to 8.7% at the larval stage. Mixtures of nuclear and messenger RNA were used to hybridize single-copy DNA and the results indicate that mRNA sequences present on neurula polysomes, but not on larval polysomes, are found in larval nuclei. Furthermore, mRNA sequences found on larval polysomes, but not on neurula polysomes, are found in the neurula nuclei. The data indicate that post-transcriptional events appear to play a role in the qualitative control of mRNA diversity during development.


Vestibular adaptation to long-term stimuli. Experimental procedures are described which respectively enlarge stimulus duration in vestibular peracceleratory tests, and allow to rule out direct thermal effects on the vestibular nerve during long term calorisations. First experimental results indicate that time course of nystagmus during prolonged stimulations differs markedly in rotational and caloric tests. Whereas there is a distinct decline of response during rotation (in accordance to the predictions of current mathematical models), in caloric tests nystagmus reaches a steady state level, maintained for at least 15 min.


[RNA-binding proteins of Rana temporaria oocytes: incorporation into informosomes of oocytes in vivo]. The participation of RNA-binding protein in the formation of informosomes in vivo was studied using an intracellular microinjection technique. The RNA-binding protein of the frog Rana temporaria oocytes was isolated by affinity chromatography and was labelled in vitro without any loss of its activity. It was shown that during cultivation of the oocytes the specific incorporation of the injected RNA -- binding [3H]-protein into the ribonucleoprotein particles occurred. These particles were further described as informosomes, characteristic ribonucleoprotein particles of animal cells.


Immunoregulatory subpopulations of human T lymphocytes. Human T lymphocyte subpopulations can be identified on the basis of surface receptors for G or M immunoglobulins (TM and TG cells respectively). Isolated TM or TG cells can be analyzed in vitro for their morphology and their functional properties. In addition to unique characteristics observed in light and electron microscopy, TM and TG cells show a different pattern of responsiveness to PHA and alloantigens. The major functional difference so far observed concerns their interaction with B lymphocytes. TM cells provide help for the T-dependent responses of B lymphocytes to pokeweek mitogen (PWM), whereas TG cells suppress B lympohcyte activation in this in vitro system by inhibiting helper TM cells. Imbalances and/or functional alterations of TM and TG cells are present in various immunological disorders and are possibly involved in the pathogenesis of these diseases.


Electron cytochemistry of peroxidase activity in neutrophils of the bone marrow of the frog Rana temporaria. The bone marrow of the frog was studied during the spring and winter periods. During the winter period, one type of large granules in the neutrophils of the frog proved to be peroxidase-positive. The remaining, smaller granules are peroxidase-negative. In the spring, immature and mature neutrophils are encountered in the frog. The mature neutrophils of the frog in the spring and winter periods are analogous. The immature neutrophils contain peroxidase activity in the perinuclear space, the endoplasmic reticulum and some of the granules. Some of the peroxidase-containing granules are directly associated with the canals of the endoplasmic reticulum. In the immature neutrophils, during the formation of the peroxidase-containing granules, peroxidase-negative granules smaller in size are already present in the cytoplasm. Thus, the sequence of granule formation usual for mammals is apparently not observed in the neutrophils of certain amphibia. "Annular" mitochondria, encompassing part of the cytoplasm were detected in the neutrophils. These apparently represent a section through a spherical mitochondrion enveloping and isolating a certain zone of the cell. It is proposed that the mitochondria in this way, differing from the lysosomal mode, participate in the catabolism of the cytoplasm.


[Effect of substances of a polypeptide nature isolated from the thymus, cerebral cortex and substantia alba on the cellular and humoral indices of immunity in thymectomized mice]. Effects of low molecular weight polypeptides (M. W. lower 10,000) isolated from the calf thymus, cortex and white matter of the brain by extraction with acetic acid on the cellular and humoral immune responses were studied in experimental thymectomized mature CBA mice. Thymectomy reduced markedly the number of T-cells in the spleen. Accordingly, the ability to generate both Ig M and IgG antibody forming cells as well as humoral antibodies to thymus-dependent antigen, SRBC, was significantly suppressed in the animals. Subcutaneous administration of 1 micron/g (body weight) of the thymus and brain cortex polypeptides during 8 days not only completely restored T-cells population in the spleen and immune responsibility but also elevated these values 1.5-2 fold in comparison with sham controls which had been given saline solution. The preparation from the white matter of the brain lacked biological activity.


["Phenomenon" of vestibular compensation]. The role of spinal afferentation from the lower part of the body in establishing compensation of the consequences of the vestibular function abaissement was studied in experiments on guinea pigs. The ligation of the spinal cord at the level of thoracic segements performed under local anesthesia neither produced appreciable effect on the compensatory development at simultaneous or subsequent destruction of the labyrinth nor destroyed it in the preliminarily labyrinthectomized animals. The ligation of the spinal cord in the labyrinthectomized animals under ether or chloroform anesthesia was accompanied by a strong disorder of the compensation. The above substances also provoked analogous decompensation in the unilaterally labyrinthectomized animals with an intact spinal cord. The results obtained indicate that the reported disorder of the vestibular compensation induced by ligation of the spinal cord under ether anesthesia is consequent on an immediate effect of inhalation anesthetics on the compensatory mechanisms rather than is resultant of abolishing spinal afferentation from the lower part of the body.


Selective absence of large forms of factor VIII/von Willebrand factor in acquired von Willebrand's syndrome. Response to transfusion. A previously healthy elderly man with mucocutaneous bleeding was found to have a benign monoclonal IgG gammapathy associated with criteria for severe von Willebrand disease (Factor VIII procoagulant activity, Factor-VIII-related antigen, and ristocetin cofactor activity, less than 10% of normal). Associated qualitative abnormalities of factor VIII/von Willebrand factor were demonstrated by radiocrossed immunoelectrophoresis and immunoradiometric assay. The late clinical onset and negative family history are in favor of an acquired form of vWD. The monoclonal gammapathy and abnormalities of factor VIII/von Willebrand factor have been stable over a 10-yr period. No inhibitor to Factor VIII procoagulant activity, ristocetin cofactor activity, or Factor-VIII-related antigen could be demonstrated. Following transfusion of cryoprecipitate (with a normal cross immunoelectrophoretic pattern), there was a rapid removal of the large forms of Factor.-VIII-related antigen, paralleled by a decay of ristocetin cofactor activity. The transfusion study of this patient with acquired von Willebrand disease type II (variant of von Willebrand disease) serves to emphasize the relationship between polydispersity of Factor VIII/von Willebrand Factor and functional heterogeneity.


Separation of functionally distinct human granulocyte-macrophage colony-stimulating factors. Human placental conditioned medium (HPCM) contans colony-stimulating factors (CSFs) required for the growth in vitro of neutrophilic granulocyte-macrophage (GM) and eosinophilic (EO) progenitor cells from human bone marrow. Fractionation of CSFs in HPCM was achieved by manipulation of the elution conditions on a column of phenyl-Sepharose. After equilibration of the phenyl-Sepharose column at high ionic strength (1 M ammonium sulfate), all of the CSF bound; one species of GM-CSF (alpha) and all of the elutable EO-CSF were eluted from the column simply by reducing the salt concentration, whereas the second species of GM-CSF (beta) was free of EO-CSF and was eluted only by increasing the concentration of tehylene glycol in the elution buffer. The two GM-CSFs were functionally distinct. GM-CSF alpha preferentially stimulated colony formation by day 14 of culture, and there was a decreased proportion of neutrophil colonies and increased proportion of macrophage colonies as the strength of the stimulus was decreased; GM-CSF beta, on the other hand, preferentially stimulated colony formation by day 7 of culture, and the proportion of neutrophil colonies was high (average 80%) and independent of the concentration of GM-CSF beta. GM-CSF alpha and GM-CSF beta were indistinguishable on the basis of apparent molecular size on tel filtration columns (molecular weight 30,000), charge properties on isoelectric focusing beds (isoelectric point, 4.9), and were not related to each other as a sialoglycoprotein is related to its asialo form. Adherent cell removal of the target bone marrow cells (to remove colony-stimulating cells) suggested that both GM-CSFs acted directly rather than by stimulating the production of GM-CSF. Mixing and titration experiments indicated that the differences in functional specificities of the two GM-CSFs (and the lack of EO-CSF associated with GM-CSF beta) were not due to the presence of specific inhibitory molecules or lower absolute levels of CSF in one fraction relative to the other. These two species of GM-CSF should be useful in separately enumerating subpopulations of different GM-progenitor cells inhuman hemopoietic disorders.


MLC-activation by acute lymphatic leukemia blasts. The MLC-activating potential of 25 ALL blasts (16 "common" ALL, 6 T-ALL, 3 not identified) was investigated. Mitomycin-treated leukemic blasts or X-irradiated lymphocytes were cultured with heparinized whole blood from different healthy donors. MLC activation by blast cells was expressed as percentage of MLC activation by X-irradiated lymphocytes. Leukemic blasts showed a heterogeneous pattern of MLC activation, ranging from 2% to 245%. Eleven out of 25 cases of ALL poorly stimulated the MLC (2% to 33% response). Twelve ALL stimulated a normal response (50% to 120%); and 2/25 ALL stimulated a supranormal response (more than 200%). Four of six cases of T-ALL stimulated the MLC as efficiently as irradiated lymphocytes, 2/6 were among the poor stimulators. Most poor stimulator blasts had, however, normal MLC-activating properties if, instead of whole blood, isolated lymphocytes were used as the responding cells. The poor activation of lymphocytes by some leukemic blasts in whole blood appeared to be associated with impaired release of blastogenic factor(s) during the MLBC. No evidence for active suppressor mechanisms was found. The significance of the MLC-activating properties of leukemic blasts for the classification and immunotherapeutic use of ALL is discussed.


Hydroxytryptamine uptake inhibitors block para- methoxyamphetamine-induced 5-HT release. 1 Activation of mycolonic twitch activity (MTA) of suprahyoideal muscle after p-methoxyamphetamine (PMA) administration in rats anaesthetized with urethane has previously been reported to be due to brain 5-hydroxytryptamine (5-HT) release. Increased MTA caused by PMA was blocked by chlorimipramine (0.1 to 1 mg/kg) and fluoxetine (0.3 to 3 mg/kg) but not by desipramine (3 mg/kg). 2 The 5-hydroxytryptophan-induced increase of MTA of suprahyoideal muscle in rats pretreated with pargyline was not blocked by chlorimipramine but was blocked by methysergide. 3 [3H]-5-HT was injected intraventricularly 30 min before the beginning of ventricular perfusion with artificial cerebrospinal fluid in urethane anaesthetized rats. PMA (mg/kg i.v.) increased the release of [3H]-5-HT in the perfusate after injection. Chlorimipramine (0.1 to 1 mg/kg) and fluoxetine (0.1 to 1 mg/kg), injected 10 min before the PMA injection, caused a dose-related blockade of the increased release of [3H]-5-HT induced by PMA. Desipramine at 3 mg/kg slightly inhibited the increased release of [3H]-5-HT caused by PMA but was inactive at 1 mg/kg. 4 It is proposed that these 5-HT uptake inhibitors block the increased MTA caused by PMA by preventing the PMA-induced release of 5-HT in the central nervous system.


The effect of potassium on frog stomach muscle. 1 KCl relaxed strips of frog stomach muscle. Usually, the effect was biphasic, i.e. contraction followed by relaxation at a concentration of 137 mM. 2 The effect was mimicked by K2SO4. 3 Ouabain (5 and 10 micrograms/ml) blocked the relaxations and reversed them to contractions. The tension of strips was not affected. 4 Phenoxybenzamine (1 microgram/ml) and procaine (20 micrograms/ml) inhibited the relaxations, potentiated the contractile component and lowered the tone of the muscle. 5 The relaxation and the tone were inhibited by papaverine (5 micrograms/ml) but the contractile component was unaffected. 6 It is suggested that potassium-induced relaxations are mediated through the activation of the sodium pump.


Ultrastructural evidence for remodelling in a central noradrenergic pathway following electrolytic lesioning. Electrolytic lesions were carried out in the medial hypothalamus of adult rats to study remodelling responses in a central noradrenergic pathway, the medial forebrain bundle. Four days, two weeks, 4 weeks and 8 weeks post-lesion, the animals were perfused and processed for correlated fluorescence microscopic (FM) and electron microscopic (EM) study. FM evaluation 4 days post-lesion showed that, compared with control preparations, catecholaminergic fibers became thick, distorted and intensely fluorescent. With increasing survival times the caliber of these fibers became finer and fluorescence intensity was gradually diminished. Some of the small blood vessels in the vicinity of the lesion acquired an intense perivascular fluorescence. Electron microscopic examination of the lesion site 4 days post-lesion disclosed many degenerating axons and increased extracellular space. No increased extracellular space was discerned by 8 weeks post-lesion. After all survival periods greatly enlarged axonal profiles were seen, and these resembled 'growth cones' described in earlier tissue culture, developmental and peripheral nervous system studies.


The filum terminale of the frog spinal cord, a non transformed preparation: I. Morphology and uptake of gamma-aminobutyric acid. The filum terminale of the frog spinal cord is a rather pure glial cell preparation, largely devoid of neuronal elements. gamma-Aminobutyric acid (GABA) is taken up by the frog filum terminale (FT) via a Na+-dependent, ouabain-inhibited, saturable high affinity transport system with a Km of 2.7 x 10(5) M. The rate of the FT GABA uptake is significantly greater than the velocities observed in the spinal cord. In fact, the Vmax increases caudally beyond the level of the last root, and is maximal in the FT per se. beta-Alanine is a competitive inhibitor of the FT high affinity transport system for GABA (Ki 11.1 x 10(-5) M). In addition to GABA, the FT also takes up beta-alanine, glycine, glutamate and aspartate at rates significantly higher than those shown by the spinal cord of the frog. Light and electron microscope level radioautography clearly shows that GABA uptake occurs primarily in the glial cells and also in ependymal cells present in the FT. In that the FT contains few ependymal cells and a large number of glia, it is fair to state that most of the GABA accumulated by the FT reflects the glial transport of this amino acid. Unlike the adult frog, the spinal cord of the tadpole does not show any regional differences in the rate of GABA transport during early development. However, during later developmental stages, the rates of GABA transport increase in the caudal portion of the tadpole cord as compared to the more rostral areas. Close to metamorphosis, the terminal portion of the tadpole cord, which is destined to become the filum terminals of the frog, accumulates GABA at rates not greatly different from those observed in the FT of the adult frog. Therefore, the tadpole spinal cord is a useful preparation in which to study the dynamic properties of normal non-transformed glia as influenced by a changing neuronal population, whereas the frog FT is a unique preparation for the study of some properties of normal glia largely in the absence of neurons.


Lessons from the study of induced alterations in amino acids in patients with cancer. The sensitivity of the neoplastic cell to amino acid deprivation shows considerable variation. The responsiveness of human leukemic cell cultures to L-asparaginase indicates that those of T cell origin are much more sensitive to the action of this L-asparagine-depleting enzyme than those of B cell origin. A cautionary note is raised concerning amino acid analogs. Minor changes such as substituting selenium for sulfur to make seleno-L-methionine rather than L-methionine may lead to considerable differences in their respective metabolic polls in vivo. A systemic survey of amino acid requirements of human neoplasms is desirable before designing new analogs.


Differential effects of trypsin on the epidermis of Rana catesbeiana. Observations on differentiating junctions and cytoskeletons. The filamentous cytoskeletons of epidermal cells of the bullfrog (Rana catesbeiana) were investigated by electron microscopy. Following treatment with trypsin, sheets of epithelium were removed from swatches of abdominal skin. Trypsinization produces differential effects on the ultrastructure of the various cell layers. The desmosomes of all layers, except those of the stratum corneum, are split by trypsinization and the resulting desmosomal plaques fastened to tonofilaments are retracted into cells to form deep "inpouchings" of the plasma membranes, while tonofilament bundles become diffuse. Epidermal sheets were gently homogenized to form a suspension of cell remnants with damaged plasma membranes as indicated by vital dye exclusion tests and electron microscopy. Cytoskeletons retain their shapes, yet the lateral distances between individual tonofilaments within bundles appear to increase, thus forming diffuse lacelike structures. These observations support the suggestion that tonofilament bundles, when fastened to desmosomes, have elastic properties. The possible role of the cytoskeletons in the maintenance of cell size and shape in an ion-transporting epithelium is discussed.


Effects of intragastric water infusion and gastric distension on hypothalamic neuronal activity. The effects of gastric water infusion and distension were examined in neurons from various parts of the rat brain. Neurons in the lateral preoptic-lateral hypothalamic-medial forebrain bundle (LPA-LH-MFB) neuropil were sensitive to gastric water infusion and distension. Cells randomly selected and examined in other brain areas were less sensitive to the same stimulation which indicated that the effects were relatively specific. The results, in terms of changes in neuronal discharge frequency from an established baseline, indicate that many cells in the lateral preoptic-lateral hypothalamic-medical forebrain bundle area are affected by intragastric water infusion and gastric distension within a time period during which changes in drinking would normally occur. These neurons were also affected differentially by acute water deprivation. LPA-LH-MFB neurons in 24 hr water deprived animals were significantly more sensitive to water infusions and less sensitive to stomach distension when compared to cells recorded in animals maintained on ad lib eating and drinking. Some of these same neurons were also sensitive to changes in the extracellular concentrations of sodium and glucose. The results are discussed in terms of the involvement of the lateral preoptic-lateral hypothalamic-medial forebrain bundle neuropil in ingestive behavior.


[Current views on pathology and new therapeutic perspective. I: Immunological aspects of the neoplasm (author's transl)]. Several experimental data support the evidence that Osteogenic Sarcoma is a virus-originated neoplasm. Cytogenetic and immunological aspects of this neoplasm are examined by the Authors on the basis of the latest advances in this field, and in view of possible therapeutic applications.


Protein A-positive staphylococci serve as a selective B cell mitogen for lymphocytes from primary immunodeficiency patients. Staphylococcus aureus protein A-positive bacteria have recently been proposed as selective B lymphocyte mitogens. We have studied the lymphocyte response to such mitogens in bacteria in normal subjects and in patients with primary immunodeficiencies. Patients with primary T cell defects show a normal response to protein A-positive bacteria and impaired responses to PHA and Con A. In contrast, patients with Bruton agammaglobulinaemia respond normally to these T cell mitogens but not to the bacteria. Thus, protein A-positive bacteria fulfil the criteria for being a T cell-independent B cell mitogen for human peripheral blood cells.


The significance of antibodies to poly(adenosine diphosphate-ribose) in systemic lupus erythematosus. Poly(adenosine diphosphate-ribose) and ds-DNA binding activity have been measured in thirty-nine systemic lupus erythematosus (SLE) sera, nineteen rheumatoid arthritis sera, fourteen sera from non-SLE rheumatic and non-rheumatic diseases and in ten normal sera. Antibodies to poly(ADP-ribose) were found only in the SLE and in three SLE-like rheumatic diseases. Anti-DNA antibodies, on the other hand, were found not only in the SLE and SLE-like diseases, but also in rheumatoid arthritis and chronic active hepatitis. Estimation of poly(ADP-ribose) binding was, therefore, more specific for, and more discriminatory of SLE from other diseases, than the estimation of ds-DNA binding. The results indicate that the estimation of poly(ADP-ribose) binding in serum may be more useful in the diagnosis of SLE than the presently employed estimation of DNA binding using the Amersham kit. DNA-anti-DNA immune complexes are detected in some of the SLE sera after deoxyribonuclease I digestion, confirming earlier reports of the existence of circulating DNA-anti-DNA complexes in SLE patients. Snake venom phosphodiesterase treatment of some of the SLE sera also resulted in increased poly(ADP-ribose) binding activity, suggesting the existence of poly(ADP-ribose)-anti-poly(ADP-ribose) immune complexes in the circulation of SLE patients. This observation raises the possiblity that poly(ADP-ribose) immune complexes may play some part in the pathogenesis of some cases of SLE.


Altered regulation of mitogen responsiveness by suppressor cells in multiple sclerosis. Suppression of the lymphocyte response to concanavalin A (Con A), phytohaemagglutinin (PHA) and protein A from Staphylococcus aureus (SpA) by Con A-induced suppressor cells was measured in twenty-four patients with recently active-recovering multiple sclerosis (MS), twelve with inactive MS and twenty-three healthy controls. Patients with recently active disease displayed significantly greater suppression of the response to Con A. Suppression of the responses to PHA and SpA did not differ among the groups. Lymphocyte stimulation in cultures not showing suppression was similar in all three types of subjects. These results suggest a disturbance of lymphocyte regulation in patients with recently active-recovering MS and illustrate the potential usefulness of measuring the suppression of responsiveness to several mitogens.


Subpopulations of T lymphocytes in myasthenia gravis patients. Subpopulations of human peripheral blood T lymphocytes were examined in twenty-three myasthenic patients. T lymphocytes bearing receptors for the Fc portion of IgG (T gamma) were significantly increased in a third of the patients examined. T lymphocytes bearing receptors for the Fc portion of IgM (Tmu) were within normal values in all but two patients. Possible implications of these cells in the pathogenesis of myasthenia gravis are discussed.


5'-nucleotidase of B and T lymphocytes isolated from human peripheral blood. The ecto-5'-nucleotidase activities of highly purified T and B lymphocytes from human peripheral blood have been investigated using biochemical and histochemical techniques. The enzyme activity of the purified B cells was about 3.5 times that of the T cells. Using a histochemical assay, 21--55% of the B cells stained positively for 5'-nucleotidase, but only 2--22% of the T cells were positive. These results are discussed in relation to the low 5'-nucleotidase activities found on peripheral blood lymphocytes from patients with chronic lymphatic leukaemia and some patients with primary hypogammaglobulinaemia.


99M Tc-PIPIDA biliary imaging in children. A report of 99mTc-PIPIDA as a diagnostic biliary agent in children is presented. The laboratory values and radiologic examinations are included to demonstrate the clinical value of the agent in evaluating both congenital and acquired pediatric biliary tract abnormalities. The role of 99mTc biliary tract imaging in children is discussed.


Computed tomography: regression of periventricular enhancing lesions in multiple sclerosis. A patient with two periventricular enhancing lesions on computed tomography subsequently demonstrated clearing of these abnormalities without treatment and a clinical course compatible with multiple sclerosis. CT scans obtained in disease processes associated with acute demyelination and alteration of the blood-brain barrier can show enhancing lesions. These lesions can initially be confused with primary or secondary neoplasms, enhancing infarcts or arteriovenous malformation. The differential diagnosis of enhancing lesions without significant mass effect should initially include the acute phase of multiple sclerosis and other demyelinating processes.


Post-operative analgesia with diflunisal. An open study was carried out in 196 consecutive patients who had undergone orthopaedic (103) or gynaecological (93) surgical procedures to assess the effectiveness and tolerability of a single dose of 500 mg diflunisal in the relief of postoperative pain. Diflunisal was given when patients first complained of pain, and pain severity before and at intervals up to 6 hours after the dose was assessed using a 10-point analogue scale. The results showed that diflunisal was both prompt and prolonged in its effect. No pain was reported after 1 hour in 138 (71%) of the 196 patients, and after 6 hours only 13 (7%) still reported some pain. Diflunisal was equally as effective in the two groups. Side-effects were reported in 13 (7%) patients but these were ones commonly found in the post-operative situation.


A trial of 5-fluorouracil and Corynebacterium parvum in advanced colorectal carcinoma. This study has confirmed that patients who have advanced colorectal carcinoma have impaired responsiveness to delayed-hypersensitivity skin testing, and also have elevated levels of serum IgM. Serial observations of delayed-hypersensitivity skin tests, total lymphocyte counts, T-lymphocyte counts, B-lymphocyte counts, and serum immunoglobulin levels failed to reveal any consistent pattern of responses in patients treated with either chemotherapy alone or chemoimmunotherapy. In 33 patients chosen at random to receive either 5-FU alone or 5-FU in combination with intramuscularly administered C. parvum, there was no evidence of objective response or influence on survival. Intramuscularly administered C. parvum, in the dose and schedule used, produced no evident immunologic or therapeutic effect.


Epidermal growth factor in the submandibular gland and serum of mice with muscular dystrophy: chemical properties in dilute gland extracts. Epidermal growth factor (EGF) has been measured in extracts of submandibular glands from mice with hereditary muscular dystrophy. RIA results show that adult male and female dystrophic mice have significantly less submandibular gland EGF than do unafflicted controls. Despite the differences in gland content of the protein, serum levels of EGF are similar in both dystrophic and control animals. Furthermore, submandibular gland concentrations of amylase are normal in the dystrophic mice, indicating that not all proteins synthesized by the glands are affected. Gel filtration studied reveal that the elution properties of EGF in extracts of glands from dystrophic and control animals are indistinguishable. Unexpectedly, the chromatographic profiles indicate that most of the EGF in gland extracts elutes as a low molecular weight protein when the molecule is studied at low, biologically active concentrations; only a small portion of the protein is associated with a high molecular weight complex. Under the same experimental conditions, submandibular gland nerve growth factor maintains its association with other components in a high molecular weight form.


Role of adherent T cells and of B cells in the immune response to purified protein derivative of tuberculin. The role of monocytes, B cells, and adherent and nonadherent T cells in the response of purified protein derivative of tuberculin (PPD) as measured by [3H]thymidine uptake in vitro has been explored. The response to PPD was found to be highly dependent on monocytes, to approximately the same extent as previously found for the responses to concanavalin A (Con A) and phytohemagglutinin. The response to PPD was also found to be highly dependent on a population of adherent T cells different from the adherent T cell population involved in the response to Con A. In the case of PPD, the effect was additive, as opposed to the potentiating effect seen for Con A. Further, the adherent T cells involved in the response to PPD were much more sensitive to hypotonic shock than those in the response to Con A. B cells were also found to be important in the response to PPD, although only to a slight extent.


The in vitro migration of murine fetal liver cells to thymic rudiments. The migration of murine fetal liver cells to thymus rudiments was studied in vitro using a migration under agar technique. There appeared to be a minor population that migrated specifically to the thymus from the age of 10 to 14 days of gestation. The specificity of migration was demonstrated in 12-day fetal liver cells by a series of competition studies. The ability of these cells to colonize a thymus rudiment was shown by further incubation after invasion of the epithelial thymus rudiments: small colonies of lymphoid cells were present in invaded tissue but absent from uninvaded control tissue. At 13 to 14 days of gestation, there appeared an additional population that migrated specifically to the spleen, as demonstrated, again, with a competition protocol. Studies with avian and human tissue as attractants in the same system showed that migration was specific to the thymus and did cross species barriers. This observation was used to demonstrate a similar attractant activity in cell-free conditioned medium from human thymus epithelial cultures, and to demonstrate the absence of such a cell-attractant factor in the conditioned medium from the thymus of a child with previously documented severe combined immunodeficiency disease.


Stimulation of sodium transport by antidiuretic hormone in bullfrog intestine. Antidiuretic hormone (ADH) increased the potential difference (PD) and shortcircuit current (SCC) across the small intestine of the bullfrog. This effect was independent of those produced by amiloride or high calcium but was masked by a theophylline-induced effect. Net active sodium (Na+) absorption accounted for the observed electrical changes.


[Structure and antipyretic-hypothermizing effect of various pyrazol/1,5-a/pyrimidines]. Structures of condensation products between 3-phenyl-5-aminopyrazole and ethyl ethoxymethylenacetoacetate and ethyl acetonoxalate are demonstrated, namely ethyl 2-phenyl-7-methylpyrazolo[1,5-a]pyrimidin-6-carboxylate and ethyl 2-phenyl-7-methylpyrazolo[1,5-a]pyrimidin-5-carboxylate. Studies on antipyretic and hypothermizing activity of the acid derived from the latter and of other compounds are reported.


Studies on the relative roles of pituitary and progesterone in the induction of meiotic maturation in the amphibian oocyte. In the amphibian, gonadotropins act on the epithelial cells surrounding the oocyte to produce and/or release progesterone which in turn acts at the oocyte surface to initiate the resumption of meiotic maturation. Since maturation is reported to require continuous exposure to gonadotropins but only brief (5--15 min) exposure to progesterone, it was of interest to reexamine the interrelationships between the two hormonal stimuli as well as the kinetics of progesterone production, metabolism, and biological activity. Germinal vesicle breakdown (GVBD) required continuous exposure to 0.005 pituitaries/ml for 6.0 h to produce 50% GVBD that occurred at 10.1 h. Actinomycin D (10 microgram/ml) completely inhibited pituitary induced GVBD when added during the first 5--6 h; 50% inhibition occurred when added at 7.3 h. Thus, actinomycin D continues to inhibit 1--2 h after the requirement for pituitary stimulation. Pituitary stimulation produced a 4-fold increase in 3H-acetate incorporation into progesterone and acetate conversion to progesterone was relatively constant during successive 2 h pulses throughout the 10 h period leading to GVBD. There was no significant metabolism of 3H-acetate derived progesterone when follicles were treated with pituitary extract, although the same follicles rapidly concentrated and metabolized exogenous 3H- or 14C-progesterone. The length of continuous progesterone exposure required for 50% GVBD varied from 11 h at 0.158 microM to less than 0.08 h at 15.8 microM. The time to 50% GVBD was only delayed by about 10% (1.5 h) when maximal and minimal progesterone levels were compared. A comparison of 3H-progesterone uptake and response (GVBD) as a function of [progesterone]0 indicated that uptake of 2--4 mumol 1(-1) cell water will induce 50% GVBD. These results indicate that a threshold uptake must be reached to initiate resumption of meiosis and that this level must be maintained throughout the period leading to nuclear breakdown. Under physiologic conditions, gonadotropins stimulate progesterone production and this progesterone is protected from, or inaccessible to, steroid metabolizing enzymes.


Sensitization of resistant myeloid leukemia clone cells by anti-cancer drugs to factor-stimulating differentiation. Studies were made on the effect of cancer chemotherapeutic drugs on in vitro differentiation of a clone (R4) of mouse myeloid leukemic cells (MI) that is resistant to inducers. Treatment of the cells with 50% ascitic fluid (an inducer) plus 0.25 microgram/ml of adriamycin or 0.3 microgram/ml of daunomycin induced phagocytic activity and suppressed cell growth, but had little effect on cell viability; treatment with ascitic fluid or the drugs alone had no effect. In combination with ascitic fluid, mitomycin-C, hydroxyurea, 5-fluorouracil, or bleomycin also induced phagocytic activity, but 6-mercaptopurine, amethopterin, or aminopterin did not. These drugs also induced other differentiation-associated properties, lysozyme activity, and locomotive activity. The present results indicate that some cancer chemotherapeutic drugs sensitive resistant leukemic cells to an inducer of cell differentiation.


Development and early prognosis of esophageal varices in severe chronic active liver disease (CALD) treated with prednisone. The prevalence of gastroesophageal varices and gastrointestinal hemorrhage was determined in 124 patients with severe chronic active liver disease (CALD) receiving prednisone and followed regularly for up to 10 yr. Varices were demonstrated by contrast radiography in only 19 patients (15%). Ten had varices before therapy and nine developed them after 12-102 mo of observation (mean, 38 +/- 9 mo.). The likelihood of developing varices within 5 yr after therapy was 8% in all patients and 13% after documentation of cirrhosis. Hemorrhage from varices occurred in only 1 patient, who had varices and bled once before treatment. Bleeding from other sites was commoner in the presence of varices (P less than 0.025), but mortality was not increased by bleeding from any site. The probability of upper gastrointestinal bleeding was only 6% within 5 yr after therapy and the 5 yr survival was 93%. In severe CALD, varices are an uncommon initial finding and do not develop quickly or hemorrhage early after steroid therapy.


Morphological identification of alpha-1-antitrypsin in pulmonary macrophages. Pulmonary macrophages, as seen in the routine sputum samples obtained from smokers, reveal bipolar orangophilic spurs. By use of histochemical and immunofluorescence techniques, these spurs have been found to contain alpha-1-antitrypsin. Morphological localization of this substance within the pulmonary macrophages may be significant.


The regulation of lymphocyte functions by the macrophage. Macrophages may exert a regulatory influence at various stages in the life of the lymphocyte - they may influence the non-antigen-driven differentiation of lymphocytes - as exemplified by the effects on thymic differentiation; they may establish the mode and form of antigen to be presented or recognized by the lymphocyte; may regulate the lymphocyte's antigen-driven functions. Each of these critical regulatory steps needs explaining in molecular terms and integrated and placed in the context of the other regulatory functions of lymphocytes. The control of secretion of MP is an eloquent example of the molecular complexities and the intricate congrol mechanisms - internal and external - operating at each step of each regulatory process. A final comment concern the question of macrophage heterogeneity. Is the same cell performing all the functions of degradation, presentation, and secretion - or cytotoxicity? Or do we have subpopulation, each with a different role? This issue is not settled. The unitarians argue that the phagocytes pass through different stages of differentiation and that each function may become more or less prominent at each stage. Certainly, the manner in which each macrophage function is assayed can condition the outcome: for antigen presentation, one adds 1 % of macrophages to cultures of spleen cells; for cytotoxic assays, the figure is 50 to 100 macrophages per tumor cell! It is our feeling that until such time as membrane molecules are identified and used as probes for differentiation or for identification of subsets we will not resolve this issue. Along these lines, macrophages have been found to have Ia antigens (Hämmerling et al. 1975, Schwartz et al. 1976) and can be divided into two sets on the basis of the presence or absence of Ia. Dorf and I have found - by cytotoxicity - that only about 35 to 50% of peritoneal macrophages bear Ia molecules (Dorf & Uanue 1977). Exceptionally, some exudates will bear up to 75% positive cells. Neither Ia-positive nor Ia-negative macrophages change significantly after prolonged periods of culture. Whether these results indicate two defined subsets of macrophages is now being investigated.


T and B cells in the transfer of immunity against Trichinella spiralis in mice. Enriched populations of T or B cells, prepared by nylon wool filtration of mesenteric node lymphocytes from mice infected with Trichinella spiralis, were capable of transferring immunity to normal and irradiated syngeneic mice. In cell recipients there was an early loss of fecundity by the worms and an accelerated expulsion from the intestine. Treatment with anti-Thy 1.2 serum, to deplete contaminating T cells, severely reduced or abolished the protective activity of enriched B-cell fractions. Replacement of contaminating T cells by normal T cells restored the capacity of B-cell fractions to reduce worm fecundity but did not result in worm expulsion. As it was shown that comparatively small numbers of T cells (3 x 10(6)) alone were effective in transferring immunity to irradiated mice it is suggested that the T cells act not as helper cells, but are involved in the generation of changes in the intestinal environment that are detrimental to worm survival.


Mast cells in severely T-cell depleted rats and the response to infestation with Nippostrongylus brasiliensis. The effect of severe T-cell depletion on mucosal mast cells of the small intestine and on connective tissue mast cells has been studied in adult thymectomized, irradiated, bone marrow reconstituted (B) rats. Under normal conditions, intestinal mucosal mast cell numbers do not differ significantly between B rats, normal age matched rats and non-thymectomized irradiated controls. Connective tissue mast cells are significantly fewer in the tongues of B rats than in normal rats, but the difference is atributable to an effect of irradiation. Infestation with Nippostrongylus brasiliensis produced approximately equal increases in mucosal mast cells in non-thymectomized irradiation controls and in normal rats. In B rats there was no increase in mucosal mast cells following infestation. B rats failed to expel the parasites normally. Failure of mast cell proliferation was not due to the effects of the persisting worm burden. Antihelminthic treatment at the time of worm expulsion by normal rats did not reveal a hitherto masked mast cell response in B rats. Nippostrongylus infestation did not reveal evidence of thymus-dependency of connective tissue mast cells. As in athymic nude mice, mucosal mast cells in the rat have been shown to be T-cell dependent during the proliferation that follows infestation with an intestinal nematode parasite.


Peanut agglutinin (PNA)-binding properties of murine thymocyte subpopulation. Surface receptors for peanut agglutinin (PNA), a lectin with D-galactose specificity, were detected on mouse thymocytes using fluorescence microscopy. Depending on mouse strain, 69-85% of unseparated thymocytes could thus be characterized as PNA+. Electrophoretic fractionation of thymocytes from normal or immunosuppressive drug-treated donors revealed an inverse relationship between PNA-binding properties and cell electrophoretic mobility (EPM). Thus, all thymocytes recovered in the lowest EPM fractions were strongly PNA+ whereas those in the highest EPM fractions were in the majority PNA-. Most of the cells collected in the intermediate EPM range were PNA+ but staining with the fluoresceinated lectin appeared weaker than for the low EPM thymocytes. Reciprocal experiments in which thymocytes were separated by PNA-mediated aggregation into fractions with different affinities for the lectin and then subjected to physical analysis, definitely established that PNA+ cells are of lower EPM than PNA- cells and that these two cell types also differ in size distribution. These data show that the four physical subpopulations of thymocytes previously described present distinctive PNA-binding properties: Th1 and Th2 cells can be classified as strongly PNA+, Th3 cells as less intensely PNA+, and Th4 cells as mostly PNA-.


Immunological senescence. II. Normal in vitro colony formation by B cells from old mice. The in vitro frequency and proliferative capacity of B cells and B-cell precursors in old mice was assessed. The mitogenic response to the B-cell mitogens lipopolysaccharide (LPS) and dextran sulphate decline later and at a slower rate than the T-cell response to phytohaemagglutinin and concanavalin A. The response to precursor B-cell mitogen dextran sulphate was not depressed in the bone marrow of aging mice. In addition, the dose response and kinetics of LPS and mercaptoethanol stimulated B-cell colony formation in agar was identical in spleen cells from young and old mice. These results indicate that the intrinsic proliferative capacity of B cells from old mice is normal.


Generation of memory cells. III. Antibody class requirements for the generation of B-memory cells by antigen--antibody complexes. Immunizing mice with preformed antigen--antibody complexes is a highly effective means of generating B-memory cells. In the present study we have compared the capacity of mouse anti-dinitrophenyl (DNP) IgM, IgG1, IgG2 and IgA antibodies to generate DNP-specific memory, when given in complex with antigen (DNP-KLH: keyhole limpet haemocyanin). Two monoclonal IgM antibodies exerted no adjuvant effect, whereas a monoclonal IgA antibody was effective, IgG2 antibodies were a more powerful adjuvant than IgG1, regardless of whether anti-DNP or anti-KLH antibodies were used. Furthermore, the capacity of DNP-KLH--antibody complexes to localize in splenic lymphoid follicles could be ranked IgG2 greater than IgG1 greater than IgA; IgM complexes did not localize in follicles. These results correlate well with data (presented elsewhere) on the capacity of these different antibodies to activate mouse complement, and confirm that C activation is an essential requirement for both follicular localization of immune complexes, and for the generation of B-memory cells. Although activation of the alternative complement pathway is sufficient to effect both processes, the results with IgG2 antibodies raise the possibility that classical pathway activation may be more effective.


Cyclophosphamide intensifies the acquisition of allergic contact dermatitis in mice rendered B-cell deficient by heterologous anti-IgM antisera. Treatment with cyclophosphamide (Cy) before contact sensitization regularly intensifies the induced sensitivity. The immunopotentiation is specific and appears due to toxicity for suppressor cells. It has been proposed that the target of Cy immunopotentiation is a suppressor B cell. We have studies allergic contact dermatitis (ACD) in mice rendered B-cell deficient by chronic treatment, beginning at birth, with a goat antiserum against mouse IgM. The ACD induced in these B-cell deficient mice was equal to that induced in intact mice. The hypersensitivity was readily and equally immunopotentiated by Cy in normal and in B-cell deficient mice. It appears that the suppressor cell that is the target of Cy immunopotentiation is not a B cell but rather a T cell.


Alveolar macrophages. III. Studies on the mechanisms of inhibition of T-cell proliferation. Unlike macrophages from the peritoneal cavity or the peripheral blood, rat alveolar macrophages actively inhibit mitogen-induced T-cell proliferation. Studies on the characteristics of this inhibitory activity revealed the following: the macrophages must be live, but mitomycin C does not block their activity; they must be added to lymphocyte cultures soon after the initiation of the cultures, and prolonged pre-incubation of macrophages in vitro diminishes their cytostatic activity; suppressive activity is most obvious in cultures of rapidly proliferating lymphocytes, and the lymphocytes themselves may be syngeneic or allogeneic; the suppressive activity may be duplicated by a low molecular-weight dialysable component of macrophage culture supernatants, and suppression of RNA synthesis is as readily demonstrable as suppression of DNA synthesis in target cells; the cytostatic effects of alveolar macrophages in lymphocyte cultures do not appear to result from target cell destruction. Studies involving repeated endobronchial lavage of rats revealed the presence of two alveolar macrophage subpopulations, one (weakly adherent in vivo) supportive to T-lymphocyte proliferation, and another (strongly adherent) strongly inhibitory; the latter population comprises the majority of alveolar macrophages in normal rats.


Characterization of the chemical and physical properties of a novel B-lymphocyte activator, endotoxin protein. Endotoxin protein, a novel mouse B-lymphocyte mitogen, is a hydrophobic acidic compound composed of approximately 85% protein and 2.2% glucosamine, but no 2-keto-3-deoxyoctonate. Endotoxin protein also contains lipid, and analysis of the fatty acids in this material demonstrated the presence of beta-hydroxymyristate, a marker for lipid A. In addition, analysis of endotoxin protein by polyacrylamide gel electrophoresis showed that it is heterogeneous, containing four or five major polypeptides, depending upon the bacterial species from which it was isolated. The mitogenicity of endotoxin protein was diminished by alkaline hydrolysis, but not by treatment with hydrochloric or acetic acid. Furthermore, its activity was resistant to digestion with trypsin, chymotrypsin, and pronase and was only partially degraded by papain.


Effect of protein deficiency on suppressor cells. The effects of moderate protein deficiency on the in vitro response of spleen cells to phytohemagglutinin in A/Jax mice were studied. The response of spleen cells from protein-deficient mice to phytohemagglutinin was found to be enhanced as compared with that of cells from control animals. Since inadequate development or function of suppressor cells in the protein-deficient mice offered a possible explanation for the enhanced lymphoproliferative activity, cocultures of spleen cells from protein-deficient and control animals were tested for their responses to phytohemagglutinin. Suppression of [3H]thymidine incorporation was detected in coculture of 25% mitomycin-treated spleen cells from control animals and 75% spleen cells from protein-deficient mice. The suppressor (regulator) elements in control spleens were found to reside in the adherent cell population.


Is DNA synthesis a requisite for the differentiation of B lymphocytes into immunoglobulin-secreting plasma cells? A great amount of conflicting evidence has been reported concerning the relationship between DNA synthesis and the differentiation of immunoglobulin-secreting cells. With the development of flow cytometry, it has become possible to analyze lymphocyte populations by making rapid and accurate measurements of cellular DNA content, and by estimating the fraction of cells that synthesize DNA. Application of these techniques to an in vitro culture system stimulated by the T-dependent sheep red blood cell antigen has allowed us to investigate cell division in populations of antibody-forming cells. DNA synthesis is apparently not a requisite for the differentiation of immunoglobulin-secreting cells.


Experimental Pneumocystis carinii pneumonia in different strains of cortisonized mice. Pneumocystis carinii pneumonia was produced in eight different strains of mice by the administration of corticosteroids, low (8%)-protein diet, and tetracycline in the drinking water. Heavier degrees of P. carinii infection were most consistently found in C3H/HeN mice; intermediate levels occurred in BALB/c AnN, C57BL/6N, B10.A(2R), AKR/J, and Swiss Webster mice; lighter degrees were found in DBA/2N and DBA/IJ mice. Histopathologically, P. carinii organisms were morphologically indistinguishable from human and rat P. carinii, and elicited a predominantly mononuclear response that was similar among the various mouse strains. The optimal cortisone acetate regimen was 1 mg injected subcutaneously twice weekly. Higher doses shortened the life span of the mice, presumably by inducing overwhelming bacterial infection. This problem occurred not only in different strains of mice, but also in the same strain of mice obtained from different breeders. Thus, cortisonized mice should be useful in the study of experimental P. carinii infection. Success of this model depends on the corticosteroid dose, as well as the strain, source, general health, and preexisting microbial flora of the mice chosen for study.


The use of hapten-polysaccharide conjugates for the induction of B-cell tolerance involving IgE responses. II. Specific tolerance induced by sulphonamide-substituted levan in the mouse. Normal and sulphonamide-sensitised mice were made tolerant with 4-sulphanilamido-benzoic-diaminopropyl-carboxymethyl-levan (4-SABA-Le). The tolerance was demonstrably specific for the sulphanilamide moiety of the hapten and extended to related compounds sharing this determinant. Mice sensitised with a 4-SABA-chicken gamma-globulin conjugate developed fatal anaphylactic shock after the injection of sulphanilamide or 4-SABA-ovalbumin, whereas allergic mice subsequently tolerised with 4-SABA-Le did not. The tolerance induced was long lasting since mice were still tolerant 7 months after an injection of an optimal dose of tolerogen. Suppressor cells play little or no part in the maintenance of this unresponsive state since even a 10-fold excess of splenic cells from tolerant animals failed to inhibit the responsivenss of 4-SABA-primed cells when they were transferred together into irradiated recipients.


Further studies on splenic material inhibiting the growth of lymphocytes in vitro. A material inhibiting growth of lymphocytes from different lymphoid organs and from different species was purified from calf spleen. A factor with a molecular weight of about 45,000 dal (estimated by gel chromatography) inhibited DNA synthesis and mitosis irreversibly in thymocytes and caused degenerative changes in the nucleus and the cytoplasm of the thymocytes as judged by electron microscopy. However, no decrease in cell number and no increase in dye uptake in a dye exclusion test were found. Growth inhibition was also demonstrated for human melanoma cells, but not for rat liver cells. The cytotoxicity and the lack of absolute lymphocyte specificity of the factor speak against its character as a lymphocyte chalone. A probably identical factor was purified from calf liver, but was not found in extracts of calf thymus.