[Multiple sclerosis in Tunisia: cochleo-vestibular study (author's transl)]. The authors undertook in Tunisia a cochleo-vestibular study of multiple sclerosis. 94 cases were included, with the following results:--67 patients had spontaneous nystagmus in the dark.--With provocation studies: - 52 cases showed signs of an irritative-type central vestibular syndrome, - 25 cases had a deficiency-type central vestibular syndrome. All of the patients underwent audition studies but the results showed that only 5 patients had a bilateral and symetrical retro-labyrinthine type perception deafness.


[Detection of circulating immune complexes in lupus erythematosus by precipitation test with polyethylene glycol and complement consumption test (author's transl)]. Immune complexe-like materials have been detected by a precipitation test with polyethylene glycol (PEG test) in the sera of 14 (93.3 p. 100) of 15 patients with active SLE, in 11 (44.0 p. 100) of 25 patients with inactive SLE, in 5 (83.3 p. 100) of 6 patients with widespread DLE and 3 (42.9 p. 100) of 7 patients with localized DLE. A good correlation was demonstrated between the value of PEG test and the complement consumption activity measured by an anticomplementary method in the cases of SLE, but it was not clear in the cases of DLE. The anticomplementary activity has been observed in the macromolecular fractions obtained by gel filtration on Sephadex G-200 of serum samples from one patient with active SLE and one patient with widespread DLE. As the value of PEG test had shown a tendency to increase with the antinuclear antibody titer if the titer was higher than 1 : 128, it is probable that the antinuclear factors were the important elements of the composition of circulating immune complexes in the cases of SLE. Contrarily, in several DLE patients, PEG test was positive despite lacking antinuclear antibody. In most cases, no significant decrease of PEG titers after DNase action was recorded, suggesting the participation of complexes containing antigens different from DNA.


Beta-lactamase effect on ampicillin treatment of Haemophilus influenzae B bacteremia and meningitis in infant rats. Intraperitoneal injections of 250 mg of ampicillin per kg every 6 h for 30 h sterilized the blood and cerebrospinal fluid of infant rats infected with either a beta-lactamase-containing strain of Haemophilus influenzae type b or a strain lacking the enzyme. However, a single injection of 100 mg/kg sterilized the blood and cerebrospinal fluid of significantly fewer of those rats infected with the beta-lactamase-producing strain. The results suggest that resistance of beta-lactamase-containing strains of H. influenzae in vivo may be inoculum dependent, as demonstrated previously in vitro. The infant rat model appears suited for the quantitative delineation of the effect of beta-lactamase on the treatment of H. influenzae bacteremia and meningitis with beta-lactamase antibiotics.


Side effects of minocycline: different dosage regimens. The incidence of side effects due to two dosage regimens of minocycline was examined over a 5-day period. A total of 60 normal women volunteers were randomly assigned in a double-blind manner to either a group who took 100 mg of minocycline twice a day or a group who took 75 mg of minocycline twice a day for 5 days. Both groups were comparable from the standpoints of age, size, race, and the use of oral contraception, nicotine, and ethanol. They were seen on a daily basis, and symptoms were evaluated by both volunteers (from diaries) and physicians. Minocycline serum concentrations were determined on blood samples taken 2 h after the a.m. dose. Volunteers taking 150 mg of minocycline per day had significantly lower serum antibiotic concentrations than those taking 200 mg per day. However, both low- and high-dose groups exhibited similar incidence and prevalence of recorded symptoms, with the single exception of nausea, where the low-dose group had fewer symptoms than the high-dose group (P = 0.035). Symptomatic volunteers did not have higher serum concentrations of minocycline than their asymptomatic counterparts. When either weight or surface area was examined with antibiotic serum concentration there was a significant inverse correlation between the two on day 2 for both groups and also on day 4 for the low-dose group. It is concluded that, in women, a dose of 150 mg of minocycline per day is associated with the same degree of side effects as a dose of 200 mg per day.


[Familial dermo-chondro-corneal dystrophy (François' syndrome)]. The François' syndrome associates disseminated firm nodular subcutaneous lesions, a deforming arthropathy and a corneal dystrophy. Inheritance seems to be recessive. Six cases of this rare syndrome have been previously published. The syndrome is reported here in two Mexican brothers. In the authors' opinion the François' syndrome is not related to the xanthomatoses. It should be considered as a genetically recessive arthropathic nodular fibromatosis.


Choice of antibiotics in management of acute osteomyelitis and acute septic arthritis in children. A survey of 158 children with acute haematogenous osteomyelitis, and of 94 children with acute septic arthritis over an 8-year period was made to determine which bacteria cause these infections. In the osteomyelitis group the organism most frequently detected was Staphylococcus aureus (74% of cases). In 16% of cases streptococci were found. Staph. aureus was also the most frequently grown organism in cases of acute septic arthritis (55% of cases), but Haemophilus influenzae accounted for 24% of positive cultures. On the basis of the survey it is the current practice of the author to use a combination of methicillin or cloxacillin and penicillin for acute haematogenous osteomyelitis, and methicilline or cloxacillin and ampicillin for acute septic arthritis. The choice of antibiotics is vitally important as treatment must start before the results of culture are known. Repeated evaluation of trends in the pattern of causative organisms is strongly recommended, in order to be aware of changing sensitivity of organisms to antibiotics.


Lymphocyte abnormalities in ankylosing spondylitis. Peripheral blood T (SRBC rosette) and B (AgG- and C-receptor) lymphocyte subpopulations and responsiveness to phytohaemagglutinin (PHA) were assayed in 40 patients with ankylosing spondylitis and in 55 normal subjects. There was no significant difference in the lymphocyte concentrations or responsiveness to PHA between the two groups. However, the percentages of T lymphocytes were significantly lower in the patients irrespective of their HLA typing. This was probably due to an increase in the 'null' population since the percentages of both the AgG- and C-receptor cells were normal.


Anti-DNA antibodies in discoid lupus erythematosus. Follow-up study. Anti-DNA antibodies have been detected previously in patients with chronic discoid lupus erythematosus (DLE) despite the absence of overt systemic manifestations. 27 patients with DLE were followed up 3 years after the detection of anti-DNA antibodies in 7. None had developed other features of systemic lupus erythematosus. We conclude that the occasional finding of anti-DNA antibodies in patients with DLE does not predict those likely to develop systemic disease.


Extraanatomical bypass of the aortic root: an experimental technique. Endocarditis involving a prosthetic aortic valve is associated with persistently positive blood cultures and aortic regurgitation. With rare exception, it is a fatal disease. An experimental technique was developed that would allow for removal of the infected aortic prosthesis with debridement and permanent closure of the aortic root. An extraanatomical outflow for the left ventricle was created using a valve-containing conduit between the apex of the left ventricle and the descending thoracic aorta (apicoaortic anastomosis). The procedure was performed on 5 mongrel dogs through a left thoracotomy without use of cardiopulmonary bypass. Four survived the procedure and required no cardiotonic support. One died as a result of a technical problem. Intraoperative pressure determination revealed a 0 to 15 mm Hg gradient across the apioaortic prosthesis and a left ventricular end-diastolic pressure of 0 to 4 mm Hg. Both postoperative angiocardiogram and postmortem examination confirmed patency of the aortocoronary bypass grafts and good function of the prosthesis.


Hyperthermic effects of D-lysergic acid diethylamide (LSD) and its derivatives in rabbits and rats. Several D-lysergic acid diethylamide (LSD) derivatives including metabolites of LSD in animals liver and Streptomyces such as D-lysergic acid ethyl, 2-hydroxyethylamide (LEO), D-lysergic acid ethylamide (LAE), D-norlysergic acid diethylamide (norLSD) and synthetic N6-allyl-D-norlysergic acid diethylamide (allyl-nor LSD) were studied with rabbits and rats. In rabbits, the order of the hyperthermic activities was allyl-norLSD greater than LSD greater than LEO, LAE, nor-LSD, and this order was parallel to that of 5-hydroxytryptamine-like activities of these compounds which was previously studied. However, the order was LAE greater than LSD greater than norLSD IN RATS. The hyperthermic effect of LSD in rabbits was attenuated by methysergide, but not by atropine when these drugs were intraventricularly administered. These results suggest the possibility that the metabolites of LSD contribute to the hyperthermic effect  of LSD in rats but not in rabbits, and support the idea that 5-HT receptors are involved in the hyperthermia induced by LSD in rabbits.


Benefits of coronary artery bypass surgery. More than 150,000 Americans have undergone coronary artery bypass surgery since this operation was first available on a nationwide scale in 1968. From data now available, we can better evaluate the various mechanisms by which bypass surgery has been thought to relieve anginal symptoms, and whether or not this operation improves ventricular function and increases survival. Most individuals have substantial or complete relief of angina after surgery, no longer require nitroglycerin or beta blocking agents, and are able to return to full physical and sexual activity. A bypass graft restores the capacity of a proximally obstructed coronary artery to increase its blood flow level up to fourfold more than resting levels with exertion. Also, evidence is now appearing that bypass surgery can improve survival in patients with advanced coronary artery disease.


Uptake of 5-hydroxytryptophan by gustatory cells in the mouse taste bud. Monoamines in the taste bud cells of the mouse circumvallate papilla were studied by fluorescence histochemistry and electron microscopy. With administration of 5-HTP (5-hydroxytryptophan) after a pretreatment with nialamide, yellow fluorescence appeared in some of the taste bud cells, while no fluorescence was observed in untreated, L-DOPA treated on serotonin treated mice. Electron microscopic study after treatment with both nialamide and 5-HTP showed small dense-cored vesicles intermingled with small clear vesicles (30-60 nm in diameter accumulated at the membranes of the gustatory cells in typical afferent synaptic contacts with nerve terminals. Definite ultrastructural change in large dense-cored vesicles (70-100 nm in diameter) could not be observed. It is suggested that the gustatory cells of the mouse take up 5-HTP and convert it to serotonin. The synaptic vesicles in the gustatory cells are believed capable of storing and releasing serotonin which presumably acts as the neurotransmitter involved in the impulse transmission from the gustatory cells to the sensory nerve fibers.


[Information transfer in the human perception of body position in space during lateral tilt (author's transl)]. (1) With eight human subjects, the perception of position in space during lateral tilt was studied by a method of absolute judgement. A preliminary analysis showed that each subject required 1000 stimuli, with 36 stimulus categories, to attain statisticach subject required 1000 stimuli, with 36 stimulus categories, to attain statistical significance. (2) Since it would be hard to experiment with such long durations, normalized experiments were pooled. The average information transfer was 3.0 bit/stimulus. (3) A closer look at the information transmission in various portions of the stimulus-response relationship (partial information transfer) revealed that near the vertical, with the head uppermost, there is more information transmission than in all other positions. This is in contrast to the visual perception of line orientation. (4) The stimulus-response relationship shows a small nonlinearity. Deviations of the body position from the vertical, with the head down, are underestimated up to 70 degrees away from the vertical; deviations from the vertical with the head uppermost are underestimated up to 10 degrees away from the vertical but overestimated up to 70 degrees.


A planned approach to coexistent cerebrovascular disease in coronary artery bypass candidates. Of a series of 874 consecutive candidates for coronary artery bypass screened for cerebrovascular disease, 49 (5.6%) were found to have significant extracarnial lesions. Combined surgical management of both carotid and coronary artery lesions was then undertaken, using a planned approach based on clinical and angiographic criteria. Cerebrovascular reconstruction was undertaken prior to establishment of cardiopulmonary bypass, utilizing staged procedures in good-risk and simultaneous reconstruction in poor-risk candidates. Our experience indicates that combined surgical treatment of the patient with unilateral carotid obstruction and coronary disease is attended by no greater risk than that attached to coronary revascularization alone. The coronary bypass candidate with bilateral carotid artery disease represents a more advanced clinicopathological state whose treatment is attended by greater risk of operative morbidity and mortality.


Inner ear degeneration in Reye's syndrome. A 9-year-old girl developed Reye's syndrome. On admission to the hospital on the fourth day of illness, responses for verbal stimuli had been absent, but were present for painful stimuli. Coma developed on the fourth day and was present until death ten days later. The fundus of both internal meatus showed vascular congestion of nerves with evidence of repeated hemorrhages. All structures within the endolymphatic system showed advanced degeneration, with exception of the tectorial membrane and cupulae. One ear showed vascular congestion, formation of thrombi, and free hemorrhage within the modiolus and spiral ganglion. In the other ear, congested vessels in nerves in the meatus were obstructed by swelling at the cribriform area. The degenerative changes corresponded closely to those produced experimentally in guinea pigs by interruption of arterial circulation to the inner ear.


Vestibular findings in 25 patients with Waardenburg's syndrome. Vestibular pathology does not seem to occur more frequently in patients with Waardenburg's syndrome than in those suffering from other types of congenital deafness. This was concluded from the results of vestibular examination by means of electronystagmography of 25 subjects with Waardenburg's syndrome from five families in the Netherlands. Ten of these subjects had a hearing loss, four of them bilateral and six unilateral. A review of the literature concerning more than 1,000 cases with Waardenburg's syndrome shows that in only 26 deaf patients was a very simple vestibular examination performed (in only three patients by means of electronystagmography).


Joint Committee on Aviation Pathology: XVII. Round window tear in aviators. It is the intent of this paper to draw attention to round window tears with the inherent symptomatology that has, for the first time, been documented in two aircrew members. It is also felt important to emphasize that the condition, regardless of etiology, could be catastrophic to the flying pilot and may be an up-recognized cause of otherwise unexplainable accidents. Not all cases of window tears cause vertigo-but some do. Not all vertigo so caused is prostrating--but some is. Because of the findings in two reported cases, it is postulated that round window membrane tear may be a significant cause of pilot disorientation. It may have been the root cause of peculiar and previously unexplained air crashes. The corollary of this hypothesis is the proposition that, in the future, careful attention be paid to the ears of pilots recovered from what would otherwise appear to be a mysterious accident.


A study of X chromosome linkage with field dependence and spatial visualization. The purposes of this report are to describe a design for the study of X linkage, to illustrate its application using cognitive test scores, and to offer a linkage hypothesis suggested by these data. Sixty-seven three-son families were examined for two X chromosome marker variables--red-green color vision and Xg(a) blood groups--and given a battery of cognitive tests of field dependence and spatial visualization abilities. Evidence was found to suggest that brothers who are identical in Xg(a) phenotype are more similar to each other in extent of field dependence than brothers who are different in Xg(a) phenotype. This result is tentative because of the small number of informative cases and the many linkage associations examined. If cross-validated, such a finding would be consistent with the proposition that an X chromosome gene contributes to the field dependence cognitive style.


Immunohistochemical identification of T-and B-lymphocytes delineated by the unlabeled antibody enzyme method. II. Anatomical distribution of T- and B-cells in lymphoid organs of nude mice. Tissue sections of the spleen, lymph nodes, and Peyer's patches were examined using an immunohistochemical demonstration of T- and B-lymphocytes in athymic homozygous nude mice possessing a known 1-5% of T-lymphocytes. Some of the few lymphocytes in the tyhmus-dependent periarteriolar zone of the spleen white pulp, the paracortex of lymph nodes and interfollicular area of Peyer's patches, were identified as T-lymphocytes. Also, singular T-lymphocytes occur in the thymus-independent marginal zone of the spleen white pulp, cortex and medulla of lymph nodes, as well as the follicular and subepithelial zones of Peyer's patches. These results indicate that the decreased numbers of T-cells in nude mice are found preferably in thymus-dependent regions, and to a smaller degree may recirculate in a similar way as in normal mice.


Isolation and characterization of membrane receptors for pokeweed mitogens from mouse lymphocytes. The major glycoproteins that bind pokeweek B-cell mitogen (Pa-1) and pokeweed T-cell mitogen (Pa-2) were isolated and identified from bone-marrow-derived lymphocytes (B-cells) and thymus-derived lymphocytes (T-cells) of C3H/He mice. The surfaces of the cells were 125I-labelled by using the enzyme lactoperoxidase, and the plasma membranes were isolated from the 125I-labelled cells. These membranes were solubilized with Triton X-100 and subjected to affinity chromatography on the affinity adsorbent prepared by coupling mitogen Pa-1 or Pa-2 to activated Sepharose 4B. The glycoproteins specifically eluted with di-N-acetylchitobiose from the affinity adsorbents were analysed according to their mobility on polyacrylamide-gel electrophoresis in sodium dodecyl sulphate. These glycoproteins were further identified by immunoprecipitation with specific antisera. Immunoglobulins, possibly immunoglobulins M and D, were identified in the eluate from the B-cell membranes, but they were not detected in the eluate from the T-cell membranes. The histocompatibility-2-complex proteins (H-2D, H-2K and Ia antigens) were found to be major receptor sites for the pokeweed mitogens on both B-cells and T-cells. However, mitogen Pa-1 (B-cell) has a stronger affinity to Ia antigens than does mitogen Pa-2 (T-cell).


[Pharmacokinetic aspects of overdosage and intoxication with oral anticoagulant drugs (author's transl)]. Three aspects of treatment with oral anticoagulant drugs are discussed: 1. Drug interactions which can lead to changes in the elimination or distribution of oral anticoagulant drugs with special emphasis on the time course of the augmentation of the anticoagulant effect. 2. Pharmacokinetic factors responsible for inter- and intrasubject differences in the response to oral anticoagulant drugs. 3. Treatment of overdosage and intoxication with oral anticoagulant drugs. Besides symptomatic treatment with vitamin K and prothrombin complex concentrate the interruption of the enterohepatic recycling by cholestyramine is demonstrated as a useful method for enhancing the elimination of phenprocoumon and warfarin from the body.


Glycosphingolipids of purified human lymphocytes. Biochemical analysis of the glycosphingolipids (GSLs) of human lymphocytes revealed qualitative and quantitative variations among purified lymphocytes from different tissues. The major neutral GSLs of tonsil lymphocytes are glucosyl ceramide (CMH), lactosyl ceramide (CDH), trihexosyl ceramide (CTH), and globoside. Thymocytes and peripheral blood lymphocytes (PBL) contain only traces of CTH and globoside, and PBL contain more CMH and CDH per cell than tonsil lymphocytes. Thymocytes and PBL contain relatively large amounts of more complex neutral GSLs that are present in only trace amounts in tonsil lymphocytes. Peripheral blood lymphocytes contained three and five times more lipid-bound sialic acid than thymocytes and toncil lymphocytes, respectively. Thymocytes and PBL contained mostly hematoside, whereas tonsil lymphocytes contained more complex gangliosides in addition to hematoside. The observed differences in GSL content among these cells may be related to their content of B cells, which comprise approximately 50% of tonsil lymphocytes, 10% of PBL and 0-2% of thymus cells, and/or the known differences in functional capacities of cells in different lymphoid organs. These findings suggest that cell surface GSLs may serve as markers for identification of functional subpopulations of human lymphocytes.


Antibodies to cytoplasmic antigens in lupus erythematosus. Serologic marker for systemic disease. Seven patients with classic cutaneous lupus erythematosus are described. Three of these patients had features satisfying four of the American Rheumatism Association (ARA) preliminary criteria for the diagnosis of systemic lupus erythematosus (SLE). Their sera, however, lacked antinuclear antibodies but demonstrated precipitating antibodies reactive against cytoplasmic RNP (La) and non-nucleic acid (Ro) antigens. Four additional ANA-negative patients lacking significant skin disease but having a lupus-like multisystem disease were found to have antibodies to soluble cytoplasmic antigens. Thirty-three of 130 ANA-positive SLE patients, but none of 16 discoid lupus patients, possessed these anticytoplasmic antibodies. These findings suggest that antibodies to Ro and La may be a marker for systemic disease in ANA-negative patients with 1) cutaneous lupus and 2) a distinct subpopulation of patients with a lupus-like syndrome without skin disease.


Circulating immune complexes and antinuclear antibodies in juvenile rheumatoid arthritis. Materials with the Clq binding properties of soluble immune complexes (IC) were found in sera from 11 of 51 consecutive (22%) children with juvenile rheumatoid arthritis (JRA) and in 17 of 20 adults with active seropositive rheumatoid arthritis (RA). IC appeared more frequently in children with systemic onset disease whereas antinuclear antibody (ANA) was found more frequently in sera from those with pauciarticular disease. Only 3 JRA sera contained anti-immunoglobulin (rheumatoid factor); those 3 also had high Clq binding activities. Seven of 50 patients (14%) carried HLA-B27 but B27 was not associated with high Clq binding activity or presence of ANA. The presence of free ANA more frequently in children with mild disease and IC more frequently in children with relatively severe disease suggests that children with systemic JRA may have a relative defect in antibody-forming capacity or reticuloendothelial function which results in decreased clearance of circulating IC. Alternatively, systemic, polyarticular, and pauciarticular JRA may represent a spectrum of clinically similar diseases resulting from different etiologic agents.


Pressure jump relaxation kinetics of frog skin open circuit voltage and short circuit current. The relaxation kinetics of frog skin open circuit voltage, Voc, and short circuit current, Isc, was studied by analyzing the effects of subjecting the tissue to sudden increments of hydrostatic pressure. Both Voc and Isc are perturbed by the pressure jump. Changes in Voc can be resolved into three components: a rapid decrease (phase I), a second, additional decrease with time constant 2.2 s (phase II), and finally a very slow increase found only in some preparations. The amplitudes of phases I and II are linear in the range of pressures studied (less than 350 atm) and have respective pressure coefficients of -1.2.10(-4) atm-1 and 3.7.10(-4) atm-1. Under short circuit conditions, phases I and II persist. The pressure coefficients of the amplitudes of phases I  and II, -4.3.10(-4) ATM-1 and -5.0.10(-4)ATM-1, respectively, are larger than those of Voc, but the time constant of phase II, 2.2 S, is the same. The sum of the amplitudes of phases I and II is directly proportional to Isc when it is inhibited with ouabain. It is argued that in both electrical states pressure perturbs the same transport mechanism giving rise to phases I and II of Voc and of Isc. The magnitude of the pressure coefficients of these processes implies that they arise from chemical reactions, rather than from simple, physical solution properties. Comparison of the pressure jump kinetics with the previous spectral analysis of the electrical fluctuations of frog skin suggests a common origin for both sets of phenomena.


Developmental abnormalities of the occipital bone in human chondrodystrophies (achondroplasia and thanatophoric dwarfism). Specific developmental malformations have been demonstrated in the occipital bone of two chondrodysplastic disorders (achondroplasia and thanatophoric dwarfism). Analysis of these malformations indicates that the occipital bone is primary affected in these disorders. In both cases, the endochondral-derived components of the occipital bone (the basioccipital, the two lateral parts, and the planum nuchale of the squama occipitalis) have failed to grow properly and are smaller and shorter than normal. On the other hand, the planum occipitalis of the squama, which derives from intramembranous ossification, is unaffected. In addition, the nature of these abnormalities indicates that the occipital synchondroses, together with the epiphyseal plates of other bones, are primarily affected in these two chondrodysplasias. The components of the occipital bone formed between the affected synchondroses failed to grow normally. The resulting malformation of the occipital bone is undoubtedly the cause of the shortening of the posterior cerebral fossa and of the considerable narrowing of the foramen magnum often described in these chondrodysplasias. It is postulated that growth disturbances between the affected occipital bone and the unaffected central nervous system results in the inadequacy of the posterior cerebral fossa and the foramen magnum to accommodate the growing brain. Consequently, compression of the brain at the posterior cerebral fossa or the foramen magnum levels could occur and thus lead to neurologic complications such as hydrocephalus and compression of the brain stem. It is suggested that the surgical removal of the fused posterior border of the lateral parts of the occipital bone (partial nuchalectomy) for the purpose of enlarging the narrow foramen magnum may be indicated in those chondrodysplastic children who develop these types of neurologic complications.


Haemobilia: a report of 2 cases.

Haemobilia: a report of 2 cases. Although rare, haemobilia should be included in the differential diagnosis of obscure gastro-intestinal bleeding, particularly when accompanied by colic and jaundice. Case histories of 2 patients with haemobilia of different origin are presented. One had traumatic haemobilia after suture of a liver rupture; in the other patient an aneurysm of a branch of the right hepatic artery ruptured into the hepatic duct. The first patient recovered spontaneously; the second required ligation of the affected blood vessel. Diagnosis and treatment of the haemobilia syndrome are discussed, with reference to the liver blood supply.


Crystalline dystrophy of the cornea: a light and electron microscopic study. We examined biopsy material from a case of stromal crystalline dystrophy of the cornea by light and electron microscopy. Frozen sections were positive for neutral fat and negative for cholesterol. Electron microscopy showed that the empty spaces at the site of the crystals did not have typical crystalline profiles. We concluded that in our case the crystals were composed of neutral fat.


Angioimmunoblastic lymphadenopathy with dysproteinemia. Three cases of angioimmunoblastic lymphadenopathy with dysproteinemia are reported which illustrate the characteristic clinico-pathological features. These cases also provide information relating to the ultrastructure of the interstitial acidophil material, the histopathological spectrum, where treatment has been attempted, and an incidence figure for an Australian community. Detailed immunological investigations in one case suggest that the disease is a primary B-lymphocyte abnormality.


Chemotherapy of cell-line-derived human colon carcinomas in mice immunosuppressed with antithymocyte serum. An in vivo model is described for assessing the antitumor activity of chemotherapeutic agents. Tumors derived from human colon carcinoma cell lines injected into antithymocyte serum (ATS) immunosuppressed mice were used. In this system, both antitumor effects and host toxicity can be quantitated, permitting calculation of a Therapeutic Index. Compared with other xenograft models, the present system is simple, experiments are completed in less than 2 weeks, and the use of cultured cell lines allows in vitro studies to be performed. The in vitro sensitivities of one colon cell line to 22 chemotherapeutic agents and of four cell lines to three agents is reported. Four drugs used in treating colon cancer (Mitomycin C, 5-FU, BCNU, and methyl-CCNU) show antitumor activity in vivo in this system. Each has a low therapeutic index. Further work with this model is indicated, with the goal of finding new drugs with high Therapeutic Indices.


Effect of sequence of administration of methotrexate, leucovorin, and 5-fluorouracil on mammary tumor growth and survival in syngeneic C3H mice. The administration of methotrexate (1mg/kg), leucovorin (1 mg/kg), and (after a 1-hr interval) 5-fluorouracil (50 mg/kg) selectively suppresses antibody production in C3H mice without inhibiting, but even stimulating, cell-mediated immunity. The effect of this regimen, given at weekly intervals, was tested on the growth of recently arising syngeneic C3H/HeJ mammary tumors. Inhibition of growth was found in three types of experiments: (a) when treatment was begun 2 days after tumor implantation, (b) when it was begun 3 weeks after implantation, and (c) when it was begun after surgical enucleation of the tumor. The extent of the effect varied from tumor to tumor, but in all cases tumor incidence or growth was significantly inhibited. Comparison was made between the above regimen and two other sequences of administration of the same drugs, which are only weakly immunosuppressive of antibody production and which do not stimulate cell-mediated immunity. Survival of tumor-bearing mice was greater with the administration of methotrexate, then leucovorin, followed by 5-fluorouracil, than it was with the other two administration schedules.


Phase II study of combined vincristine, adriamycin, cyclophosphamide, and methotrexate with citrovorum factor factor rescue in metastatic breast cancer. Fifty patients with metastatic breast cancer were treated with 8-day courses of vincristine (1 mg iv, Day 1), adriamycin (50 mg/m2 iv, Day 1), cyclophosphamide (100 mg/m2 orally, Days 1-8), methotrexate (200 mg iv by 3-hour infusion, Day 8), and citrovorum factor rescue (15 mg in 12, 18, and 24 hours after methotrexate, Day 8) at 3-4-week intervals. Forty-two patients had previously received treatment with hormones and 17 patients had received chemotherapy. Fifteen patients achieved a complete remission (CR) and 24 patients a partial remission (PR). There was a significant correlation between the response and the number of metastatic organs (0.01 less than P less than 0.02). The response rate was roughly uniform irrespective of the organ predominantly involved (0.2 less than P less than 0.3). The remission duration was significantly longer for the patients with CR compared with that for the patients with PR (0.001 less than P less than 0.01). The patients who achieved CRs and PRs survived significantly longer than the patients with no change and progressive disease (P less than 0.001). The toxic effects of the treatment were acceptable and no drug-related deaths occurred.


Correlation between local oxygen tension in muscle tissue and survival time in tourniquet shock. Tourniquet shock was induced in rabbits by constricting one hind leg for 4 hr. Local pO2 on muscle surface was measured by a multiwire platinum electrode according to the method of Kessler. Untreated animals showed anoxic tissue areas after a period of 30 min of reperfusion. All these animals died in shock within 3-4 hr. The glucocorticoid, prednisolone (20 mg/kg), and the proteinase inhibitor, Trasylol (20,000 KIU/kg), were administered 5 min bfore release of the tourniquet. These animals showed normal pO2 values during the 2-3 hr observation period. Tourniquet shock was prevented and survival time prolonged by these agents. This protective effect could be correlated to the oxygen tension in the ischemic region.


Myocardial consequences of coronary artery bypass graft surgery. The paradox of necrosis in areas of revascularization. Myocardial infarction after coronary artery bypass graft (CABG) surgery has been described clinically in up to 30% of patients but there is little morphologic information about the character and pathogenesis of the myocardial injury. We studied myocardium in the distribution of bypassed and nonbypassed coronary arteries for the presence of contraction band necrosis as compared to coagulation necrosis, in 58 autopsied patients who died less than 1 month after surgery. Operation related necrosis consisting of focal subendocardial contraction band necrosis was present to some degree in 48 (83%) patients. Regional transmural necrosis was present in 22 (38%) patients and was of two types. Contraction band necrosis occurred in 18 patients and was in the distribution of a patent bypassed coronary artery in 15 of them. Coagulation necrosis was found in four patients, and in each was in the distribution of a new graft-releated coronary artery occlusion. The results suggest that coronary artery reflow through widely patent grafts following the period of operative nonperfusion, rather than graft or intrinsic coronary artery occlusion, accounts for the majority of operation-related myocardial "infarcts" associated with CABG surgery. Thus, prevention of intraoperative myocardial injury must also focus on characteristics of the phase of myocardial reperfusion.


Mechanisms of cellular enzyme release. I. Alteration in membrane fluidity and permeability. Above-normal plasma enzyme activities resulting from increased release of intracellular macromolecules are an important diagnostic feature of Duchenne muscular dystrophy. These and other biochemical and histological characteristics of this disease are faithfully duplicated when imipramine and serotonin are administered to the rat. Imipramine, but not serotonin alone, causes release of enzymes from rat diaphragms and human lymphocytes and release of hemoglobin from erythrocytes in vitro. Ouabain causes neither. Imipramine-induced enzyme release is decreased by adding ATP to the specimen in vitro or in hypertonic solution. Imipramine inhibits the capping phenomenon (an aggregation of antigen/antibody complexes of the membrane) of human B lymphocytes labeled with fluorescein-conjugated antihuman immunoglobulins. Serotonin alone has no such effect, but, administered together with imipramine, it potentiates the inhibition of capping by imipramine.


Direct spectrophotometric determination of alpha-amylase activity in salive, with p-nitrophenyl alpha-maltoside as substrate. We describe a simple, direct kinetic method for determination of salivary alpha-amylase (1,4, alpha-D-glucan 4-glucanohydrolase, EC 3.2.1.1). The assay makes use of a well-defined substrate, p-nitrophenyl alpha-maltoside, which is hydrolyzed by alpha-amylase to a chromogenic product, p-nitrophenol. Activity is determined by directly monitoring the increase in absorbance of the reaction mixture. Amylase activity can be defined in international (IUB) units of micromoles of product/min per liter of saliva. For 22 healthy subjects, the mean +/- SD of amylase activity in mixed saliva was 2.77 +/- 1.12 U/liter. Activity and instrumental response were linearly related over the entire range tested (0.224 to 11.90 U/liter). The within-run precision (CV) over this range was better than 3% for all but the lowest activities. Values obtained with this assay correlate well with those obtained with a modified Nelson-Somogyi saccharogenic method (r = 0.979). The precision and simplicity of this assay suggest that it is the method choice for determining amylase activity in human saliva.


[Prognosis of acute upper gastrointestinal bleeding before and after the introduction of emergency endoscopy (author's transl)]. A total of 416 patients had been admitted to hospital for acute upper gastrointestinal bleeding, about half of them before the introduction of endoscopy. There was a highly significant decrease in total mortality rate by 18 percent since the introduction of endoscopy. The reason for this is probably largely the much higher number of correct identification or localisation of bleeding points by endoscopy (98 percent) when compared with conventional radiological examination.


[Small-nodular liver cirrhosis with marked portal hypertension due to vitamin A intoxication resulting from psoriasis treatment (author's transl)]. Clinical and histological signs of small nodular liver cirrhosis with portal hypertension were present in a 36-year-old man, three-and-a-half years after a seven-week course of treatment of psoriasis vulgaris with high doses of vitamin a (70 X 10(6) IU orally). Although there is no increase in serum level of vitamin A now, increased deposits of vitamin A in the perisinusoidal lipid storage cells (Ito cells) are still demonstrated by fluorescencespectrophotometry and under the electron microscope. Fundectomy with resection of the terminal oesophagus was necessary because of bleeding from oesophageal varices.


[Treatment of chronic pain with intracerebral stimulators]. A new technique for the functional treatment of chronic intractable pain with a cerebral stimulation system under the patient's control is described. A four-pole electrode, diameter 0.65 mm (made by Medtronic), was implanted under stereotaxic control into nine patients. Stimulation was via a substernally implanted receiver connected to the intracerebral electrode. A pocket-sized stimulator with circular aerial, intermittently placed on the patient's skin, was activated by the patient whenever pain occurred. Pain suppression was achieved in all instances, best results being obtained with stimulation of the medial mesencephalic lemniscus, including the sensory-motor thalamic nuclei. The pain-suppressing effect lasted for up to seven hours, so that three stimulations for 30-40 minutes daily were sufficient, as demonstrated by an observation period of up to 21 months. The only complication was transitory oculomotor paresis. The described reversible non-destructive stereotaxic and functional technique of stimulation is preferable to the coagulation method in the treatment of chronic intractable pain.


[DDAVP: alternative to replacement treatment in mild haemophilia A and von Willebrand-Jürgens syndrome (author's transl)]. 1-Deamino-8-D-arginine vasopressin (DDAVP), a vasopressin analogue, increases factor VIII levels in plasma. A female carrier of haemophilia A with known bleeding diathesis and markedly reduced factor VIII activity was successfully treated with DDAVP during bilateral Caldwell-Luc's operation for chronic maxillary sinusitis. An estimated 5000 U factor VIII concentrate was calculated to have been saved. DDAVP is the first alternative to replacement therapy in the treatment of moderate and mold forms of haemophilia A and von Willebrand's disease.


Isolation and characterization of presynaptically acting neurotoxins from the venom of Bungarus snakes. 1. Five presynaptic toxins have been isolated in pure form from the venom of Bungarus multicinctus and Bungarus caeruleus and named beta1, beta2, beta3, beta4, and beta-ceruleotoxin. 2. They differ in electrophoretic mobility and amino acid composition, while all have the same molecular weight (22000) and are composed of two subunits of molecular weight 9000 and 12000. 3. The toxins have phospholipase A activity when assayed with both natural and synthetic phospholipids, and this activity requires the presence of Ca2+ ions. 4. beta-Bungarotoxin (beta3) binds 1 mol of Ca2+ per mol of protein and this binding induces a conformational change as detected by fluorescence measurements in the presence of the dye 8-anilino-1-naphthalene sulfonic acid. 5. The phospholipase activity of all the toxins is lost when a critical histidine residue is modified with p-bromophenacyl bromide. 6. As a result of the modification the lethality of the toxins is greatly reduced. 7. Native toxin causes a rapid decrease in amplitude of end-plate potentials, followed by a transient increase and subsequent decrease, until transmitter release is completely abolished. The modified  toxin still causes the early decrease in release but toxin action does not progress to complete block. 8. The rate of blockage of transmitter release by native toxin is reduced in the presence of modified toxin. 9. It is concluded that phospholipase activity plays an important role in the action of this class of toxins at the neuromuscular junction.


Solution conformational analysis of 2'-amino-2''deoxyadenosine, 3'-amino-3'-deoxyadenosine and puromycin by pulsed nuclear-magnetic-resonance methods. The solution conformation of 2'-amino-2'-deoxyadenosine, 3'-amino-3'-deoxyadenosine, and 3'-amino-3'-deoxy-6-N,N-dimethyladenosine have been determined by nuclear magnetic resonance in aqueous and ammonia solutions. The analysis of the ribose moiety is based on the two-state S in equilibrium N model of Altona and Sundaralingam. Longitudinal proton relaxation time and nuclear Overhauser enchancement measurements have been carried out in order to characterize the orientation of the base relative to the ribose. Those studies indicate that 3'-amino-3'-deoxyadenosine and 3'-amino-3'-deoxy-6-N,N-dimethyladenosine exist in solution preferentially in the N-anti-g + conformations. On the other hand, 2'-amino-2'-deoxyadenosine adopts the S-syn-g +/t conformation families. It appears that the base is restricted to the anti conformation in the first two compounds, while in 2'-amino-2'-deoxyadenosine, one third of the molecules in the S state are in the anti range. These studies corroborate the previously proposed correlations between the N state of the ribose and the anti orientation of the base and between the S state of the ribose and the syn orientation of the base.


Human B cells alloantigens; separation from other membrane molecules by affinity chromatography. Human Ia-like alloantigens have been solubilized from membranes of B lymphoblastoid cell lines using sodium deoxycholate (DOC). They have been purified by affinity chromatography using specific rabbit antibodies bound to an agarose column, eluting the antigens at pH 11.0 in the presence of 0.5% DOC. The isolated, purified, material contained two proteins of molecular weights 35 000 and 27 000 by sodium dodecyl sulfate gel electrophoresis, apparently noncovalently associated with each other. The molecules were completely separated from the soluble products of the HLA-A, B and C loci and retained serological activity as measured by their capacity to inhibit the lysis of B lymphoblastoid cell lines by B cell-specific alloantisera.


Preliminary characterization of a glycoprotein having Fc receptor properties extracted from a T cell lymphoma (L-5178-Y). After incorporation of 14C-labeled amino acids and/or of [3H]fucose during in vitro culture of the Thy-1.1-bearing, Fc receptor-positive T lymphoma, L-5187-Y, attempts were made to purify the Fc binding structure(s). Following solubilization of a crude membrane pellet using sodium deoxycholate, the 110 000 x g supernatant was filtered on Sephadex G-200, and the fractions containing IgG-binding material were further purified by affinity chromatography (on IgG Sepharose 4B.) Sodium dodecyl sulfate polyacrylamide gel electrophoretic analysis of the final Fc-binding product showed an apparent molecular weight of 110 000 daltons. On subsequent reduction with 2-mercaptoethanol, five bands (mol. wts. 56 000, 36, 000, 25 000, 18 000 and 15 000) were observed, the latter two being probably degradation products. These results are in accord with some of the published data concerning B cell Fc receptors.


The effect of chlorpromazine on cell membrane resistance and capacitance. Chlorpromazine, at concentrations of 1-50 micron, caused rises of up to 10 mV in the p.d. across the isolated frog skin and decreases as great as 5% in membrane capacitance; at concentrations of 10 micron and above the resistance decreased, by up to 25%. Effects on p.d. and resistance were obtained only when the drug was added to the solution bathing the outside of the skin; these effects are possibly caused by increases in the sodium permeability of the outer membrane. The decrease in capacitance is not consistant with an increase in membrane area without changes in thickness or permittivity, but could be explained by a loss of water from the membrane.


Clonazepam and 5-hydroxytryptophan-induced myoclonic stereotypy. The effect of clonazepam on a behavioral model of increased whole brain serotonin activity was investigated. While clonazepam induces a moderate elevation of whole brain serotonin, the drug fails to potentiate or inhibit 5-hydroxytryptophan-induced stereotypy in young guinea pigs. These results suggest that despite drug induced alterations in serotonin concentration clonazepam does not hava an effect on the physiologic activity of serotonin and may not exert its pharmacologic activity by influencing serotonin within the brain.


Metabolic pathways of delta-aminolaevulinic acid in Rhodopseudomonas spheroides. Incorporation of delta-aminolaevulinic acid 5 14C and 4 14C into the inosine monophosphate pool and into porphyrins, was studied in cell suspensions of R. spheroides. The results contradict a direct incorporation of delta-aminolaevulinic acid into the purine ring of nucleotides through gammadelta-dioxovaleric acid. It would suggest a nonspecific incorporation after degradation of delta-aminolaevulinic acid without a transamination as a first reaction.


Ultrastructural autoradiographic study of blast cells in the mouse thymus. Interest for radioleukemia research. Ultrastructural autoradiographic studies of mouse thymic blast cells after H3 Tdr injection show that their fine nuclear structure is related to their position in the cell cycle. The variations in the composition of the subcapsular blast cell population during radiation-induced leukemogenesis indicate kinetic changes in thymic lymphopoiesis, which are probably due to the oncogenic process.


Steps in mitogenic action of concanavalin A. I. Colchicine- and cytochalasin B-sensitive events. The role of colchicine-sensitive and CB-sensitive events in Con A-inducible activation of spleen cells and thymocytes was investigated by exposing the cells to varying concentrations of the drugs, separately or in combination, in serum-free or serum-supplemented medium and at different stages of the culture. In the initial phase, thymocytes were much more sensitive to colchicine than spleen cells. In adequate doses, either colchicine or CB alone could completely inhibit the activation process; when both drugs were combined, the resulting effect was, as a rule, not additive. It was possible to separate the delayed spontaneous agglutination (which is a concomitant of the mitogenic effect of Con A) from activation by selectively inhibiting the former with CB. It was thus concluded that the delayed agglutination is not a prerequisite for activation to be accomplished. Following a low dose of CB which is known to inhibit cell mobility without blocking activation, agglutinates cannot form spontaneously, but do so when cell collisions are produced by mechanical forces.


[Effect of pancreatic kallikrein and trypsin inhibitors on the course of postischemic toxemia]. By studying the ECG findings, dynamic changes of the body temperature, the edemas size in the tourniquet-compressed limbs, vascular permeability of the skin, general condition and survival rate on a model of post-ischemic toxemia in albino rats it was found that the serum containing antibodies to pancreatic kallikrein (200 gamma/ml), when introduced parenterally (1 ml per 100 g of the mass), increases the survival time of the animals and prevents the development of gangrene in the ischemic limbs. The curative effect of the antikallikrein serum resembles the action of ingitril, a polyvalent inhibitor (1.5 Un. per 100 g of the mass). The antitryptic serum employed in analogous doses stands by its action close to the normal rabbit's serum.


[The treatment of a case of postpartum uterine atony with consumption coagulopathy by therapeutic pharmaco angiography (author's transl)]. Two hours following a twin stillbirth at 30 weeks gestation, a 25 year old primipara developed severe postpartum hemorrhage due to uterine atony and complicated by increasing consumption coagulopathy. Bilateral catheterization of the internal iliac arteries was attempted but only achieved on the left. For the next 22 hours the patient received 0.2 units per minute of Pitressin by intra-arterial infusion. The infusion was done more and more intermittently as the bleeding stopped and the coagulation parameters normalized. An emergency ligation of the internal iliac arteries or an emergency hysterectomy was thus avoided in this patient.


[The influence of pregnancy on the immunologic reactivity of the mother. Investigations into the reactivity of the peripheral lymphocytes to mitogens and on the percentage rate of B-cells in the maternal blood during pregnancy (author's transl)]. In pregnant women, lymphocytes from the peripheral blood were stimulated with phytohemagglutin (PHA), pokeweed mitogen (PWM) and Concanavalin A (CON A) and the percentage rate of B-cells was determined. Compared to healthy non-pregnant controls of the same age, no significant difference in the stimulation or the percentage of B-cells was found at different times of pregnancy. The observation of other authors was confirmed that phytohemagglutin induced stimulation of lymphocytes in non-pregnant controls can be diminished by the addition of pregnant serum instead of the normally used homologus AB serums. Our results lead to the conjecture that the diminished reactivity of the maternal lymphocytes during pregnancy is not intrinsic to the lymphocyte in pregnancy but more likely due to inhibitory substances in the serum of the pregnant patient.


[Pharmacological studies of ketoprofen (19583RP) III. Anti-inflammatory, analgesic and antipyretic actions in subcutaneous administration (author's transl)]. It has been already reported that ketoprofen (KP) has a potent anti-inflammatory action comparable to indomethacin, but with oral administration, gastric mucous membrane disturbances occur. In the present work, we administered the Na salt of KP (KP-Na) subcutaneously and found that the anti-inflammatory action was more potent in subacute and chronic inflammations than in the acute one. As an acidic compound, KP-Na had a relatively potent analgesic-antipyretic action and a medical efficacy comparable to the usual non-steroidal anti-inflammatory drugs given orally. The efficacy in case of subcutaneous administration was 2 to 3 times stronger over both acute and chronic inflammations than in case of oral administration. On the other hand, the gastric mucous membrane disturbance was decreased to about 1/3 in case of subcutaneous administration, thus the gastric disturbance could be abated and the medical efficacy can be increased when KP-Na is given subcutaneously. As KP-Na was less irritative at the injection site, the compound could be used clinically for a subcutaneous administration.


Alpha-1-antitrypsin phenotypes in Lyon, France. Alpha-1-antitrypsin Pi phenotyping was performed by thin-layer isoelectric focusing on samples from 1653 healthy white blood donors. The variants were confirmed by the acid-starch gel technique and crossed immunoelectrophoresis, with complete agreement between the two methods. The allele frequencies in this population were PiM, 0.9019; PiS, 0.0713;PiZ,0.0142; PiI, 0.0036; PiF, 0.0036; PiV, 0.0024. In addition, some rare phenotypes (MX, IS, LM) were noted. No difference was noted in the distribution of the variant alleles between males and females. The significance of the high frequency of the S allele is related to the ethnic origin of this population. The agreement of the results obtained by our technique and by acid-starch gel electrophoresis confirms the validity of Pi determination by thin-layer isoelectric focusing.


Preparation of high titered rabbit anti-mouse IgE sera. A method for the consistent production of rabbit anti-mouse IgE sera is described. Rabbits were immunized intravenously with IgE-anti-normal mouse serum mixtures over 8 to 12 weeks. Ten microliters of the anti-mouse IgE sera completely neutralized all the mouse IgE in the serum (50 microliter) possessing a passive cutaneous anaphylaxis titer of 20,000.


Adenosine deaminase activity during the growth cycle of T and B lymphoid cell lines. Adenosine deaminase activity per cell was constant during the growth of B cells but increased during logarithmic growth of T cells and decreased as T cell viability decreased during the stationary phase of growth. This elevated enzyme activity appears to be a biochemical marker for T cell leukemic blasts.


Effect of iodoacetate on T lymphocytes of young AKR mice. The influence of sulphydryl inhibitor iodoacetate on properties characteristically associated with T-lymphocytes was examined in young AKR mice. Thymocytes from mice receiving 100 microgram of iodoacetate showed a decrease in cortisone sensitivity, and responded more vigorously to phytohemagglutinin (PHA). Spleen cells from treated mice also exhibited greater reactivity to both PHA and concanavalin A (Con A). The uptake of 14C-iodoacetate by the thymus, spleen, lymph nodes, bone marrow, liver and kidneys also was investigated. One hour, 2 h and 18 h after the intraperiotoneal injection of 10-3 M iodoacetate, the thymus demonstrated a higher incorporation of the labelled compound than did the spleen, lymph nodes or other organs examined. These findings provide further evidence to indicate that the immuno-enhancement observed following iodoacetate treatment is related to its effect on T lymphocytes.


Anti-DNP antibody response after the topical application of DNFB in mice. A single painting or daily paintings for 5 days with dinitrofluorobenzene (DNFB) on the abdominal skin of mice induced both contact sensitivity, detectable by ear swelling, and, hapten-reactive helper T cells, detectable by the augmented anti-bovine serum albumin (BSA) antibody response on challenge with dinitrophenyl-BSA. Contact sensitivity was induced within 7 days and helper activity within 14 days after the sensitization. Anti-hapten antibody response in the spleen or regional lymph nodes of such mice, however, was negligibly small during the 15 days after a single painting. Failure to respond with anti-hapten antibody production of mice given only a single painting was shown to be due to the shortage of B cells reactive to the hapten. Daily paintings for 5 days did not necessarily result in the augmented antibody response. By contrast, a strong anti-hapten antibody response was observed in mice receiving two paintings at an interval of 10 days. In these mice, hapten-specific B memory cells as well as hapten-reactive T cells were detected. Thus, the anti-hapten antibody response after the topical applciation of simple chemicals may depend upon the priming of B cells, and the response must be mediated by the cooperation of T and B cells both reactive to the antigen.


Trypanosoma rhodesiense infection in B-cell-deficient mice. B-cell-deficient C57B1/6J mice (suppressed from birth with goat anti-mu) and controls (treated from birth with normal goat serum) were infected with Trypanosoma rhodesiense. There was a significant (P less than 0.01) decrease in duration of survival of the mu-suppressed mice compared with that of controls. Whereas both mu-suppressed and control mice had an initial rise in parasitemia of similar magnitude, only the control mice exhibited a subsequent period during which the parasitemia fell to undetectable levels. In control mice, immunization with irradiated organisms prevented the development of detectable parasitemia after challenge with viable trypanosomes. However, immunization did not alter the course of infection in B-cell-deficient mice. These results indicate that immunity to T. rhodesiense infection in mice is dependent on B-cell immunocompetence.


Surface glycoprotein patterns of normal and malignant human lymphoid cells. I. T cells T blasts and leukemic T cell lines. Exposed surface glycoproteins of resting and in vitro activated human T lymphocytes and leukemic T-cell lines were labelled by the galactose oxidase-tritiated sodium borohydride method. The labelled glycoproteins were separated by polyacrylamide slab gel electrophoresis and visualized by autoradiography. The basic glycoprotein patterns of the T lymphocytes and T blasts were found also in the leukemic T cells. The glycoprotein pattern of the T-cell lines was easily distinguishable from that of other hematopoietic cell lines. The findings suggest that: (1) surface glycoprotein analysis might be useful for the identification of cell lines and for the differential diagnosis of hematopoietic malignancies; and (2) cells of cultured T lines may be used for the identification and preparation of T lymphoid differentiation antigens and perhaps also tumor-associated surface molecules.


Effects of daunomycin and radiation on cell-survival and repair of DNA single-strand breaks. The combined action of Daunomycin and irradiation was investigated using mouse L-929 cells in culture. Survival of cells was measured with the colony assay. Sedimentation in alkaline sucrose gradients was used to study repair of DNA single-strand breaks (SSB) in the presence of various concentrations of Daunomycin. A small increase in radio-sensitivity, as measured by decreasing Do, was obtained for doses of Daunomycin that are considerably toxic to the cells (0.1 microgram/ml). However, the Dq values remained constant even at high concentrations indicating that Daunomycin does not interfere with recovery processes. The rate of rejoining of SSB remained constant up to 1.0 microgram/ml, whereas concentrations of Daunomycin as high as 10 microgram/ml reduced the velocity of repair by a factor of 13. Our data show that concentrations of Daunomycin similar to those required for other DNA-binding drugs are required to inhibit SSB repair. For clinical purposes, no increase in tumour-killing efficiency may be expected from a combined treatment with Daunomycin and radiation.


gamma-radiolysis of N2O-saturated aqueous solutions of trehalose and trehalose/amino acid mixtures. gamma-radiolysis of aqueous N2O-saturated solutions of alpha, alpha-trehalose (10(-2) M) with doses of 500 krad yielded glucose (G = 2.3), gluconic acid lactone (G = 0.25) and--after reduction--iditol (G = 0.15), mannitol (G = 0.05), 5-deoxy-glucitol (G = 0.34) and 2-deoxy-glucitol (G = 0.14). The decomposition of trehalose (G = -5.9) was reduced, if equimolar amounts of amino acids (alanine, leucine, phenylalanine, methionine or cysteine) were present during irradiation. The extent of this reduction has been correlated with the .OH radical-scavenging properties of the added amino acids. Cysteine also protected trehalose by the repair of initially formed trehalose radicals and almost completely suppressed the formation of products. The addition of the remaining amino acids led to an increase of molar product yields (glucose, 5-deoxy-glucitol and 2-deoxy-glucitol), which was related to the decomposition of trehalose. This finding was explained by hydrogen transfer from the amino acids to precursor radicals of the products.


Pulse radiolysis of lactate dehydrogenase. Pulse radiolysis has been used to investigate the rates and transient spectra for the reactions of free radicals with beef heart lactate dehydrogenase at pH 7. Analysis of the results leads to second-order rate-constants for eaq-, .OH, .I, .Br2-, .I2- and .(CNS)2- which are, respectively, 24, 21, 10, 0.55, 0.43 and 0.15 in units of 10(10) M-1 s-1 with uncertainties of +/- 20 per cent. Those for .I and .I2- are similar to the corresponding rate-constants for the related enzyme alcohol dehydrogenase. The spectra of the transient species produced by .OH, .Br2- and .(CNS)2- all showed evidence for reactions with tyrosine and tryptophan residues, and in general terms the magnitudes of the rate-constants appeared to increase with the oxidizing abilities of the radicals. The implication of the results for understanding the mechanism of deactivation by free radicals is discussed.


Change in radiosensitivity of sea-urchin eggs during early cleavage stages. III. Effects of split-dose irradiation with 137Cs gamma-rays. When sea-urchin eggs were irradiated with 137Cs gamma-rays, their radiosensitivity, expressed by the percentage which formed pluteus larvae, fluctuated during the early cleavage cycle. Split-dose irradiations were made both in the sensitive and resistant phases. For eggs in the sensitive phase, the effect of the first exposure of 500 rad was not diminished during the interval before the second exposure. Eggs irradiated in the resistant phase were only slightly damaged. Results imply that fluctuations in radiosensitivity of sea-urchin eggs are caused mainly by different degrees of non-repairable damage in each phase of cleavage rather than by different recovery abilities.


Cellular immunity in patients with leprosy. Circulating T lymphocytes and their response to PHA in leprosy. The ability of peripheral blood lymphocytes from polar lepromatous (LL), borderline lepromatous (BL) and borderline tuberculoid (BT) patients to transform in vitro in the presence of phytohemagglutinin was found to be significantly reduced. A significant reduction in the percentage and absolute number of T lymphocytes was observed in LL cases. In BL cases the number of T lymphocytes was decreased, but the reduction was proportional to the reduction in the total lymphocyte population was observed only in bacteriologically positive cases.


The effects of X-radiation  on thymidine-transport kinetics and DNA synthesis in Ehrlich ascites tumour cells in relation to extracellular thymidine concentration. A TdR carrier-transport system, believed to be facilitated diffusion, has been shown to exist in Ehrlich ascites tumour cells. It is suggested that this system is the predominant transport mechanism at low extracellular concentrations (less than 1-5 micron). The transport system was damaged considerably by 5 krad X-radiation, resulting in a 30-35 per cent reduction in the  initial total TdR uptake rat at low extracellular concentrations and 15-20 min after irradiation. The extent of the damage was dependent on the age of the cells as was reflected by relative decreases in V max and Km. It can be concluded that the enhanced depression in 14C-TdR incorporation into DNA of irradiated cells when low precursor concentrations were used for monitoring, is partly attributed to the radiation-induced damage to the carrier-transport system. The permeability constant for passive diffusion in asynchronous E.A.T. cells and the endogenous natural rate of dTTP synthesis in S-phase cells were estimated.


Free radical scavenging in protection of human lymphocytes against chromosome aberration formation by gamma-ray irradiation. Human peripheral blood lymphocytes were exposed to 300 rad gamma-rays, in the presence or absence of radical scavengers, and the change in the frequency of chromosome aberrations was analysed with attention directed to the protection by scavengers against the formation of primary damage leading to chromosome aberrations. The results showed that the damage involved in the formation of exchange-type aberrations was efficiently protected by scavengers, and about 60 per cent of them resulted from indirect action that could be abolished by alcohols. The SH-compounds afforded additional protection. The comparison of protective ability with reaction rates demonstrated that the indirect effect was due to the reaction of OH radicals possibly to DNA as a target molecule, and the involvement of H and eaq-was unlikely. In contrast to the exchange-type aberrations, terminal deletions were not significantly protected, suggesting that the damage leading to the terminal deletion differed in its nature from the leading to the exchange-type aberration.


Heat sensitivity of Haemophilus influenzae containing defective prophage. Strains of Haemophilus influenzae that carry a defective prophage are more sensitive to heat than is a strain that does not, even in the presence of a rec-1 mutation, which normally renders prophage noninducible. The prophage of HP1c1, a nondefective phage, does not affect the heat sensitivity.


Ceruminous tumor of the jugular foramen. Intrapetrous ceruminomas are rate tumors; only four cases have been reported in the literature. These had characteristics of cerebellopontine angle tumors. Our case had the characteristics of a tumor of the jugular foramen.


An analysis of transport, exchange, and binding of sodium and potassium in isolated amphibian follicles and denuded oocytes. The uptake and efflux of 22Na and 42K were studied in fully grown, prophase-arrested Rana pipiens follicles and denuded oocytes. Follicles and denuded oocytes contained large stores of both cations, although only 15--30% of the total Na+ and 1--2% of the total K+ pools exchanged within the 12-h period studied. 22Na uptake and efflux in follicles exhibited two-step kinetics with a smaller rapidly exchanging fraction (12--15%, t0-5 approximately 15 min), and a larger slow fraction (t0-5 approximately 0-5 day). Denuded oocytes displayed a single kinetic fraction with a rate constant similar to the slow fraction of follicles, suggesting that the fast fraction was associated with the follicular epithelium. Both follicles and denuded oocytes exhibited fast and slow 42K uptake kinetic fractions, although the follicular fast fraction was considerably larger. Conversely, whereas follicular 42K efflux also proceeded with two-step kinetics, 42K efflux from denuded oocytes appeared as a single slow fraction. Thus, a large portion of the fast K+ fraction taken up by the follicles is attributable to the somatic cells of the follicular envelopes. Na+ and K+ activities were measured in the cytoplasm of the in situ (follicular) oocyte using cation-selective microelectrodes. After correcting for Na+ and K+ associated with the follicular envelopes, only about 6% of the total ooplasmic Na+ and less than 1% of the K+ could be accounted for as bound or sequestered.


[Immediate and late results of 171 therapeutic portal systemic by pass operations (author's transl)]. The authors reports the results of 171 cases of portal systemic by pass operations out of a total of 200 cases operated on between 1968 and 1974. These 171 patients all presented with a syndrome of portal hypertension with oesophageal varices and all had previously bled. Their age lay in 70 p. cent of cases between 40 and 60 years and the cause of hypertension was in 96 p. cent of cases an intra-hepatic block. 41 p. cent were included in the risk group A according to Child's classification, 51 p. cent in Group B and 8 p. cent in Group C. The routine operation was side-to-side portacaval anastomosis (75 p. cent). The operative mortality was 5.2 p. cent in all with 1 p. cent in cases with risk A, and 15 p. cent in cases with risk C. The fall in portal pressure was on average 15 cm of water, i.e. 41 p. cent of the initial pressure. The overall survival after 5 years was 65 p. cent, 70 p. cent for risk A and 26 p. cent for risk C. In 69 p. cent of cases the cause of death was liver failure. Encephalopathy, studied in 76 patients over an average period of 3 years, occurred in 39 p. cent of cases, and in 13 p. cent of the latter it was serious. In the 12 p. cent of survivors, we noted recurrent hemorrhage, but in only 2.5 p cent of cases did the bleeding definitely come from oesophageal varices. In the light of these results, the authors judge positively the surgical treatment of portal hypertension.


The cytoarchitecture of the torus semicircularis in the Tegu lizard, Tupinambis nigropunctatus. The torus semicircularis (TS) of the Tegu lizard extends from the superficial caudal mesencephalon, dorsal to the exiting trochlear nerve, to a position ventral to the middle part of the optic tectum and its ventricle. It has an oblique orientation with the caudal pole abutting the midline while the rostal end is lateral and slightly ventral. The TS consists of a central nucleus and several adjacent cell groups. The central nucleus and the laminar nucleus, situated medially, extend the entire length of the TS while the cortical nucleus, situated dorsally and laterally, is present only in the caudal superficial portion. The central nucleus is composed of ovoid neurons with branched, radiating dendrites. The dendrites are directed medially and laterally with spines on the distal portion of the dendritic tree. The laminar nucleus consists of three to five neuronal layers. It is mainly composed of fusiform neurons with one dendritic trunk from each extremity of the soma. There is little branching and few dendritic spines. The cortical nucleus is a laminated region consisting of alternating layers of neurons and lateral lemniscal fibers. The neurons of the superficial layers are fusiform with their long axis perpendicular to the long axis of the brainstem. They possess two main dendritic trunks which parallel the laminae and are covered with dendritic spines. The deeper layers consist of pyramidal neurons with three dendritic trunks, secondary branches, and few spines. The long axis of these neurons extends from the center of the TS to the periphery. Two dendritic trunks extend dorsally or laterally towards the surface, while the third extends towards the central nucleus. The dendrites, thus, extend across the laminae. In addition, a cell-free lateral zone is described.


The cerebellum of the bullfrog tadpole (Rana catesbeiana). The cerebellum of the premetamorphic bullfrog tadpole differs from the cerebellum of other frog species in its morphology and maturational state. The cell mass beneath the floor of the lateral recess and bordering its lateral wall that has been reported to form the auricular lobe in other species is absent, the auricular lobe abutting the medial wall of the lateral recess instead and is continuous with the corpus cerebelli. The corpus cerebelli, although immature and yet to acquire an external granular layer, is already massive and displays an incipient molecular, Purkinje cell and granular layers. Cytodifferentiation in the auricular lobe and corpus cerebelli is similar, their constituent cells being in various stages of development. Fully mature cells are absent, but a small population of Purkinje cells and glia in the auricular lobe and along the marginal zone of the corpus cerebelli show advanced development. The orientation of these Purkinje cells is parallel to the pia and appears to approximate the course of the vestibulo-lateral commissural fibers. In the ventral part of the corpus cerebelli, developing climbing fibers are present but Purkinje cells are poorly developed.


Reward versus performance in self-stimulation: electrode-specific effects of alpha-methyl-p-tyrosine on reward in the rat. The hypothesis that alpha-methyl-p-tyrosine (AMPT), an inhibitor of catecholamine synthesis, reduces brain stimulation reward was tested, using a measure of reward previously shown to be relatively unaffected by variables that alter performance but not reward. The rewarding effectiveness of stimulation was determined by the location of the sharp rise in the function relating running speed in an alley to the number of pulses received as a reward. For some electrodes, AMPT depressed self-stimulation performance (speed of running) without producing any sizable effect on the measure of reward (location of rise). For other electrodes, the rewarding effectiveness of the stimulation was greatly reduced by AMPT and restored by L-dopa. These opposing results could be repeatedly demonstrated on different electrodes in the same rat. The electrode-specific differential sensitivity to AMPT suggests neurochemically disparate substrates for reward.


The usefulness of color vision testing and color fields. Routine color vision testing to differentiate between hereditary and acquired color deficiencies and the usefulness of color fields.


Separation and partial characterization of fractions derived from frog lung homogenates. A possible marker system for amphibian pulmonary surfactant. The purpose of this study is to determine if inframammalian vertebrate (amphibian) lung contains certain nonspecific esterases that have been identified as enzyme markers for mammalian (rat and mouse) pulmonary surfactant. Density gradient centrifugation procedures were utilized to concentrate any surface-active material in frog lung homogenates. Lipid and protein analyses of one of the derived fractions and of pulmonary lavage fluid were consistent with other techniques indicating that these preparations were surface active. A comparison of the nonspecific esterases in the derived fractions and the pulmonary lavage fluid allowed the identification of a nonspecific esterase that has an electrophoretic mobility comparable to one of the nonspecific esterases already identified as an enzyme marker for mammalian (rat and mouse) pulmonary surfactant. These results indicate that these enzyme markers may be useful in the further investigation of the surfactant systems of other inframammalian vertebrates.


Catecholamine innervation of the forebrain in the bull frog, Rana catesbiana. The innervation of forebrain catecholamine (CA) were experimentally investigated with use of sensitive fluorescence method of glyoxylic acid formaldehyde in the brain of the bull frog, Rana catesbiana. The CA of the olfactory bulb is supplied by CA neurons situated in olfactory bulb. And CA neurons in the hypothalamus contribute the main source for the forebrain CA except olfactory bulb. The hypothalamic CA neurons also give rise to long descending axons to innervate the brain stem. Judging from their anatomical aspects it seems that the structure homologous to mammalian nigro-neostriatal dopamine or mesolimbic dopamine system is not present in amphibian brain.


Comparative anatomy of the topography of catecholamine containing neuron system in the brain stem from birds to teleosts. The distribution of catecholamine (CA) containing neurons and nerve terminals in the brain stem from birds to teleosts were studied by means of histofluorescence technique of glyoxylic acid-formaldehyde combining method. The present study revealed that there exists marked differences in topography of CA neurons between higher vertebrates (birds and reptile) and lower vertebrates (amphibia and teleosts). Tegmental CA neurons in the birds and reptile are much more developed than those of amphibia and teleosts. Furthermore, huge collection of CA neurons (presumably dopamine) in the midbrain tegmentum could not be found in the amphibia and teleosts. On the other hand, distribution of CA nerve terminals in the brain stem is essentially similar throughout the vertebrate brains, though there exist modifications by secondary pattern distortions which correspond to various conditions of their lives. Some possible significance of CA neurons on development of central nervous system was discussed.


[The activities of the transmitter metabolizing enzymes monoamineoxidase and acetylcholinesterase and of succinic dehydrogenase were investigated in the telencephalon of Salmo irideus (teleostei) (author's transl)]. The activities of the transmitter metabolizing enzymes monoamineoxidase and acetylcholinesterase and of succinic dehydrogenase were investigated in the telencephalon of Salmo irideus (teleostei). There is a high reaction intensity of the MAO in the whole telencephalon with graded differences between the various nuclear areas. It is highest in the area dorsalis telencephali; the larger fiber tracts (Tr. olfactorius medialis et lateralis; medial and lateral forebrain bundle) are strong MAO-positive, too. There is an extremely weak AchE-reaction throughout the telencephalon. The only nuclear area which is clearly histochemically demonstrable is the regio Dm2 in the area dorsalis telencephali. The SDH-activity is demonstrable in the whole telencephalon. The activity is higher in the area dorsalis as compared with the area ventralis telencephali. In the ependym there are only a few traces of MAO- and SDH-reaction products. The recent findings may support, from a histochemical view, the hypothesis that the telencephalon of Salmo irideus represents brain structures which show structural and functional similarities with limbic telencephalic structures of higher vertebrates.


Vaginal colonization with Corynebacterium vaginale (Haemophilus vaginalis). Vaginal cultures for Corynebacterium vaginale and confidential questionnaires were obtained from unselected young women who consulted a gynecologist in a student health service. In all, 466 women were studied, 150 (32.2%) of whom were colonized with C. vaginale. Logit analysis defined four factors that were significantly associated with colonization with C. vaginale: nonwhite race, use of oral contraceptives, no history of marriage, and a history of pregnancy. Sexual experience had little influence on colonization; C. vaginale was isolated from 16 (29%) of 56 sexually inexperienced women and from 40 (41%) of 98 women who had had intercourse with six or more men. After a few patients with trichomoniasis were excluded, there was no association between colonization with C. vaginale and an abnormal vaginal discharge, either as reported by the participant or as noted by the examining physician.


Rapid loss of factor XII and XI activity in ellagic acid-activated normal plasma: role of plasma inhibitors and implications for automated activated partial thromboplastin time recording. Rapid prolongation of the aPTT of normal plasma upon incubation with ellagic acid containing aPTT reagents was observed. The aPTT prolongation was not due to time-dependent changes in pH in the incubation mixture or loss of activity of the labile coagulation factors VIII and V but occurred as a result of rapid progressive inactivation of ellagic acid-activated factors XII and XI. Prolongation of the aPTT and loss of contact factor activities was not observed in plasma incubated with particulate activator reagents. This finding seemed to indicate that adsorption of factors XII and XI to larger particles during the activation process might protect these factors from inactivation by naturally occurring plasma inhibitors. Evidence is presented which supports previous findings that C1-INH, alpha1-AT, and antithrombin (in the presence of heparin) contribute to factor XIIa and XI a inactivation in ellagic acid-activated plasma and that plasma albumin may compete with factor XII for ellagic acid binding. The data indicate that ellagic acid-containing aPTT reagents have unfavorable properties which seriously limit their usefulness in the clinical laboratory, particularly in respect to recording of the aPTT with certain fully automated clot timers.


Structure-activity relationships of corticosteroid feedback at the hypothalamic level. Structure-activity studies on the corticosteroid fast and delayed feedback receptor mechanisms controlling the secretion of corticotrophin releasing factor (CRF) were carried out with the rat hypothalamus in vitro. The secretion of CRF was induced by acetylcholine (3 pg/ml). The fast feedback receptor appears highly specific, and the structure essential for efficacy involves an 11beta-hydroxyl group and an unblocked 21-hydroxyl group. Several steroids showed antagonism and so the binding site is not very specific. 18-hydroxy, 11-deoxycorticosterone, progesterone, 17alpha-hydroxyprogesterone and 11-deoxycorticosterone were antagonists of fast feedback. The delayed feedback receptor required either an 11beta-or a 21-hydroxyl group for efficacy. The binding site required a 17-hydroxyl group when the 11beta- or 21-hydroxyl groups were absent. Binding also involved the 3-oxo,4,5-ene structure since steroids in which these are absent were inactive.


Regulation of B-lymphocyte clonal proliferation by stimulatory and inhibitory macrophage-derived factors. A functional subpopulation of murine B lymphocytes proliferate in semisolid agar culture in the presence of 2-mercaptoethanol to form colonies. The effects of diffusible macrophage-derived factors on this focal proliferation was investigated using a two-layer culture system which prevented macrophage-lymphocyte contact and permitted B-cell activation to be critically assessed under conditions of extremely low cell densities. Adherent peritoneal macrophages incorporated within underlayers of spleen or lymph node cell cultures potentiated both the number and size of developing B-cell colonies. These effects were most striking when low numbers of spleen or lymph node cells, or macrophage- depleted lymphoid cell suspensions were used. Thus, macrophage-depleted lymph node ceils gave rise to virtually no colonies, but colony-forming ability was restored by the presence of an optimal number of macrophages. When the number of macrophages exceeded that required for optimal stimulation, colony formation was suppressed; an effect which was largely prevented by indomethacin, an inhibitor of prostaglandin synthesis. Under these conditions, stimulation and inhibition of B-cell activation by macrophages could be dissociated, indicating that each signal is selectively controlled by individual molecules elaborated by the macrophage. With an appropriate number of macrophages required for B-cell activation, and sufficient indomethacin to inhibit the accumulation of macrophage-derived prostaglandin, B-lymphocyte clonal proliferation was a linear function of the number of B cells placed in culture. In the absence of macrophages, B-cell colony formation was potentiated by both lipopolysaccharide and intact sheep erythrocytes through a mechanism different from that of the macrophage-derived stimulatory factor. In addition to their direct stimulatory effect on B-cell proliferation, lipopolysaccharide and sheep erythrocytes were each capable of modulating the production and/or release of B-cell stimulatory and inhibitory factors by the macrophage. Parallel studies of conventional mitogen- stimulated lymphocyte cultures did not show a requirement for macrophages and confirm that the semisolid assay is uniquely suited to studies on the regulatory role of the macrophage in B-cell activation.


Enrichment of specific suppressor T cells and characterization of their surface markers. A simple method is described which allows antigen-specific suppressor T cells to be enriched by greater than 100-fold. The enriched cells have the following characteristic markers: Ig-, Thy-1+, Ly-1-, Ly-2,3+, and I-J+. More than 30% of this population could be killed directly by an antiserum specific for the I-J subregion gene product in the presence of complement.


Role of IgD in the immune response and tolerance. I. Anti-delta pretreatment facilitates tolerance induction in adult B cells in vitro. Adult spleen cells from C57BL.Ige mice, which generally are resistant to in vitro tolerance induction in the B-cell compartment, became hyporesponsive (tolerant) when cultured with antigen in the presence of an anti-allotype serum. Both antigen and anti-delta had to be present for this effect, which was hapten-specific and did not occur in C57BL/L mice, which lack the Ig5-1 allotype of the delta-chain detected in this system. Preculture with anti-mu serum plus antigen, in contrast, did not cause tolerance induction in adult spleen B cells of either strain. These results suggest that the surface IgD may act as a failsafe receptor to prevent tolerance induction in adult B cells. Tolerance studies with spleen cells from mice with markedly reduced numbers of IgD+ve cells, because of regimen of repeated injections of anti-delta serum beginning at birth (delta-suppressed mice), confirmed the importance of membrane IgD in preventing tolerance, because such delta-suppressed mice were hypersusceptible to tolerance by antigen alone. Inasmuch as immature B cells lack IgD on their surface, these studies suggest that acquisition of IgD is an important maturational step in the ability of murine B cells to discriminate tolerogenic and immunogenic signals.


Antigen mediation of a late-acting suppressor T-cell activity. Carrier-specific suppressor T cells can suppress antibody secretion by high avidity IgG plaque-forming cells (PFC) within 90 min in vitro. This process can be blocked by the inclusion of soluble carrier in the cell mixture or by the exposure of target cells to anti-carrier antibodies or pronase. Moreover, suppression can be augmented by PFC exposure to the soluble hapten-carrier conjugate. Finally, carrier specificity may be overcome by preincubation of the target population with a hapten-heterologous carrier before addition of heterologous carrier ATC. Thus, it is likely that high avidity suppression depends upon immunogen bound to the surfaces of antibody-secreting cells which serves as a target for suppressor cells or molecules.


B-cell activation by lipopolysaccharide. Distinct pathways for induction of mitosis and antibody production. The role played by macrophages in two effects of lipopolysaccharide (LPS) on the immune system of the mouse-substitution for helper T cells and induction of B-cell mitosis-has been investigated. C3H/HeJ mice are unresponsive and do not produce (as other strains do) antibody to 2,4,6-trinitrophenol (TNP) conjugated with autologous mouse erythrocytes (MRBC-TNP) in the presence of LPS. We found that C3H/HeJ spleen cells produce antibody to MRBC-TNP when (a) LPS and macrophages from LPS-responsive C3HeB/FeJ mice or (b) tumor necrosis serum ([TNS] induced by LPS in responsive mice) are added. The mitotic response was not restored. The findings suggest that adjuvanticity and mitogenicity represent distinct pathways of B-cell activation by LPS, subject to different regulatory mechanisms.


Immunological unresponsiveness to thymus-independent antigens: two fundamentally different genetic mechanisms of B-cell unresponsiveness to dextran. The immune response of mice to the alpha-l-6 epitope of dextran (Dx) B512 was found to be under genetic control. The congenic mouse strains A, A.CA, A.SW, A.TH, and A.TL exhibited a specific defect in their response to alpha-l-6. Also strain CBA/N was unresponsive to alpha-1-6, but the mechanism of unresponsiveness was found to be different. Unresponsiveness to alpha-l-6 in congenic A strains was not due to suppressor cells. Although these strains failed to respond to the alpha-l-6 epitope, they responded strongly to the hapten Fluorescein isothiocyanate (FITC) conjugated to Dx, indicating that the Dx can function as an efficient carrier in these strains. Dx was a potent polyclonal B-cell activator in congenic A strains as well as in high responder strains. Polyclonally-activating concentrations of lipopolysaccharide (LPS) failed to induce the synthesis of anti-alpha- l-6 antibodies in congenic A strains, although antibodies of all other specificities studied were produced. However, in high responder strains, LPS induced the synthesis of anti-alpha-l-6 antibodies. It was concluded that congenic A strains do not express V genes coding for antibodies against alpha-l-6. In contrast, strain CBA/N failed to respond to both the alpha-l-6 and FITC epitope on Dx, whereas they could respond to FITC conjugated to horse erythrocytes. Dx induced a very small, if any, polyclonal antibody response in B cells from CBA/N mice or male CBA/N x DBA hybrids, whereas Dx was a very potent polyclonal B-cell activator in female hybrids. It is concluded that CBA/N mice are nonresponders to Dx or haptenated Dx, because the cell population that can respond to the polyclonal B-cell activating properties of Dx is severely depleted.


B-lymphocyte Fc receptor-associated non-H-2 antigens are determined by a single polymorphic locus which is linked to the Mls locus. Certain non-H-2 alloantigens are associated with murine B-lymphocyte Fc receptors in that pretreatment of spleen cells with alloantibodies against these antigens inhibited binding of Ig complexes to B-cell Fc receptors. This inhibition was specific in that: (a) as has been shown previously, the Fc portion of the alloantibody was not required to produce inhibition; and (b) antibodies against some non-H-2 antigens but not antibodies against others (including some that were expressed on B cells) caused such inhibition. Backcross experiments revealed that the B-cell Fc receptor-associated non-H-2 antigens were determined by the gene(s) of a single background locus in each of the three strains tested (A/J, B10, and CBA/J). This locus was poly-morphic in that at least four different B-cell Fc receptor:associated non-H-2 antigens were identified (one each in the A/J, B10, and CBAJJ and one antigen shared or crossreactive between the B10 and CBA/J). These antigens were primarily but not exclusively expressed on B lymphocytes as determined by immunofluorescence studies, and on the basis of capping experiments they did not appear to be identical to B-cell Fc receptors. Linkage studies revealed that the locus which determined these antigens was not linked to the albino locus nor the heavy chain allotype locus and expression was neither sex-limited nor an X-linked recessive trait. However, this locus was closely linked but not identical to the Mls locus (apparent recombination frequency 6.8 percent). Thus, two closely linked non-H-2 loci both determine the expression of antigens which have characteristics similar to Ia antigens. One locus is polymorphic and determines the expression of antigens which are primarily expressed on B cells and are specifically associated with the Fc receptors of these cells. The other (Mls locus) determines antigens which are stimulatory in mixed lymphocyte cultures. These observations suggest that there may be a second gene complex which is the analogue of the I region of the H-2 complex.


B-cell tolerance. IV. Differential role of surface IgM and IgD in determining tolerance susceptibility of murine B cells. During ontogeny IgD appears later than IgM on splenocytes of neonatal mice (1) and at a time when mice develop a markedly increased immune responsiveness (2). Based on these observations, it was suggested that IgD serves as a "triggering" isotype for induction of immune responses, whereas surface IgM functions as a tolerizing receptor (3). To test this hypothesis, the susceptibility of adult splenocytes (which are predominantly mu(+)delta(+)[4-6]) and neonatal splenocytes (which bear predominantly IgM [mup(+); 1, 4-6]) to tolerance induction were compared. The results indicate that neonatal splenic B cells responsive to thymus dependent (TD) antigens are exquisitely susceptible to tolerance induction compared with those from adult mice (7-9). However, cells from both adult and neonatal mice were highly susceptible to tolerance induction when thymus independent (TI) antigen was used as immunogen (8). These results suggest that the major precursor for the TD response is a mu(+)delta(+)-cell which appears late in ontogeny and is resistant to tolerance induction and that the mup(+)-cell is the major precursor for the TI response and is highly susceptible to tolerance induction. Other differences between responders for TI and TD antigens have been described previously (10-12). To test this concept, adult splenocytes were treated with papain under conditions in which IgD, but not five other surface molecules, was removed (13). Such treated splenocytes were shown to be markedly susceptible to tolerance induction, resembling TD responders from neonatal animals. This experiment was interpreted as indicating that IgD confers resistance to tolerance induction on mu(+)delta(+)-cells. To prove this interpretation, it is necessary to show that specific removal of IgD with anti-delta also results in increased susceptibility to tolerance induction and that treatment with anti-mu does not have a similar effect. In the present studies, we have removed surface IgM or IgD by antibody-induced capping and assessed the tolerance susceptibility of the treated cells. Our results demonstrate that removal of IgD, but no IgM, from TD responders increases their susceptibility to tolerance induction.


Lymphocyte culture: induction of colonies by conditioned medium from human lymphoid cell lines. The presence of phytohemagglutinin or pokeweed mitogen in cultures of human peripheral blood mononuclear cells in agar is known to stimulate the formation of lymphoid colonies. We now report that similar colonies can be induced in the absence of plant lectins upon addition of filtered and ultracentrifuged conditioned medium (CM) obtained from certain human lymphoblastoid cell lines. Colony formation required at least 6 X 10(5) mononuclear cells per milliliter, and optimum results were obtained at concentrations of 1 X 10(6) cells/ml in the presence of 20% CM (50-500 colonies per 10(6) cells cultured). Individual cells within colonies displayed uniform morphological characteristics of lymphoid cells, and the majority formed rosettes with sheep erythrocytes, suggesting that they were of T-cell type.


Frog lysozyme. V. Isolation and some physical and immunochemical properties of lysozyme isozymes of the leopard frog, Rana pipiens. Frog Lysozyme has been purified by sequential application of acid extraction, salt fractionation, CM-cellulose chromatography, heat treatment, and gel filtration. Eight isozymes of purified lysozyme were found to be stable during prolonged storage. Isozymes were separated by preparative polyacrylamide gel electrophoresis, Ninety percent of the lytic activity of frog ovarian egg was represented by forms 7 and 8, the most highly charged isozymes. Seventy-eight percent of frog liver lysozyme activity was that of form 4. Forms 7 and 8 differed from form 4 by being larger (apparent molecular weight of 18,000 vs. 16,000), by remaining active in more acidic environment, and by exhibiting a dependency upon NaCl for activity. Antiserum prepared against frog form 4 did not react with frog forms 7 and 8 and antiserum to chicken egg-white lysozyme did not react with any frog lysozymes. All frog lysozymes showed identical reversible binding to deaminated chitin. Apparent size differences and lack of immunological cross-reactivity suggest that at least some of the isozymes are non-allelic.


Interpretation of plantar reflexes: biasing effect of other signs and symptoms. Twenty neurologists were asked to judge a number of plantar responses on film. Each film was preceded by a slide with a fictitious abstract of history and examination (minus the plantar reflex). The main part of the presentation only served to disguise the fact that two films, both showing equivocal toe movements, were presented twice at the same sitting, but with opposing information as to the probability of a Babinski sign. Interpretation of these identical pictures differed significantly (P less than 0.01), conforming to the information given. Thirty other neurologists who rated the films without previous data showed no such change of opinion.


Microtubules associated with postsynaptic 'thickenings'. Using a new albumin technique, microtubules can be seen closely related to or associated with the postsynaptic 'thickening' of mature and immature central nervous synapses. Thus in conventionally fixed synapses (without albumin pretreatment) where microtubules cannot usually be observed running into the postsynaptic dense material, this material could perhaps, in part, consist of the debris of in vivo microtubules. Smooth endoplasmic reticulum (ER) is often seen associated with the microtubules near the postsynaptic 'thickening'. Microtubules, and possibly smooth ER, may have an important role in the initiation of synapse formation and in the maintenance of mature synapses.