Evidence for corticotropin releasing factor (CRF) synthesis in the preoptic nucleus of Xenopus laevis tadpoles: a preliminary report based on lesion experiments. For the study of the hypothalamo-hypophysial system of Xenopus laevis tadpoles, hypothalamic lesions were made by means of the electrocoagulation technique. Lesioning of the ventral region of the preoptic nucleus resulted in a decrease of the number of ACTH cells in the pars distalis of the pituitary gland and in a diminution of the PAS-positive reaction of these cells. In addition, regeneration of the neurosecretory cells of the ventral region of the preoptic nucleus observed 6 weeks after lesioning was accompanied by the reappearance of normal PAS-positive ACTH cells in the pars distalis. It is suggested that the neurosecretory cells of the ventral region of the preoptic nucleus of Xenopus laevis tadpoles are related to the ACTH synthesizing cells, probably by producing CRF.


Electrophoretic variation of serum alpha1-antitrypsin treated with neuraminidase. Purified or crude sera of several different phenotypes of alpha1-antitrypsin were treated with neuraminidase. The mobility of the alpha1-antitrypsin zone was delayed on starch gel electrophoresis following incubation. Treated, M-type alpha1-antitrypsin migrated in a manner similar to the untreated S type. Treated S migrated in a similar way to the untreated Z, and treated Z did so more slowly or cathodally. After exhaustive treatment, alpha1-acidglycoprotein bands appeared near the usual M or F positions. Treated samples still possessed the antigenic activity of alpha1-antitrypsin. The concentration and trypsin-inhibitory capacity were not changed by treatment. Several evaluations and considerations are discussed concerning the metabolism of some types of alpha1-antitrypsin.


Chromosomal breaks in T and B lymphocytes in Fanconi's anemia. Three patients with Fanconi's anemia were analyzed for chromosome breaks. T and B cells were separated and grown in tissue culture with PHA and pokeweed antigen to ascertain the rates of breakage in these lymphocytic subpopulations. It has been found that there is no statistically significant difference in breakage rates in T and B lymphocytes. It is postulated that both T and B cells could be involved in the development of leukemia in Fanconi's anemia patients, assuming that chromosome breaks constitute a factor predisposing to the development of malignancy.


C3 metabolism in a patient with deficiency of the second component of complement (C2) and discoid lupus erythematosus. A patient with a hereditary deficiency of the second component of complement and discoid lupus erythematosus with features of systemic lupus erythematosus was studied. The propositus had a 9-year history of rash and arthralgia. Transient renal disease had completely resolved; there was a history of seizures. Examination of his serum disclosed antinuclear antibodies but no total haemolytic complement activity. C2 was absent. Serum concentrations of C1s, C3, C5 and C9 were elevated; other complement components were present in normal concentration, including C3 pro-activator. The patient's C3 pro-activator was electrophoretically converted by inulin and four of five lipopolysaccharides, but was poorly converted by aggregated human IgG. Two separate turnover studies with radiolabelled C3 showed fractional catabolic rates of 3-03 and 2-48% of the remaining plasma pool/hr (range of three normals: 1-62-2-18%/hr); and estimated C3 synthetic rates of 2-74 and 2-31 mg/kg/hr (range of three normals: 0-89-1-40 mg/kg/hr). Serum complement profiles of the patient's family demonstrated that the C2 deficiency was inherited as an autosomal codominant. One sibling, homozygous for C2 deficiency, and three other siblings, both parents and one daughter, all heterozygous for C2 deficiency, are in good health. Immunofluorescent studies of the patient's diseased skin exhibited substantial deposits of IgG, IgM, C1q, and C4 but not of later acting complement components, properdin, or C3 proactivator. These studies do not support the notion that inflammation in C3-deficient individuals with lupus erythematosus is mediated by the alternative complement pathway.


Cellular aspects of selective IgA deficiency. Five patients with no detectable serum IgA (less than 20 mug/ml) and one patient with low serum IgA were compared to normal subjects. The number of circulating E-RFC was normal as was the lymphocyte DNA synthesis induced by PHA, Con A, and streptokinase-streptodornase. The patients had normal numbers of IgA-bearing lymphocytes and normal or increased numbers of B cells. Purified anti-immunoglobulin antibodies specific for IgG, IgA and IgM induced a normal lymphocyte DNA synthesis as did PWM. The patients' lymphocytes were able in vitro to transform into actively secreting IgA plasmocytes. This transformation was determined by counting the IgA and immunoglobulin-containing cells and then measuring the IgA and IgG secretion in the cultures. In some patients PWM was selectively suppressive in IgA B-cell transformation into IgA secreting cells; in the other patients PWM had no effect on the IgA B-cell differentiation. PWM enhanced the IgG secretion in the patients' cultures as well as IgA and IgG secretion in the normal controls.


T and B lymphocytes and Reed-Sternberg cells in Hodgkin's disease lymph nodes and spleens. Lymphoid cells from twenty-four untreated Hodgkin's disease biopsies were examined for spontaneous sheep erythrocyte and sensitized ox erythrocyte rosette formation for the identification of T cell and cells with Fc and C3 receptors and surface immunoglobulin. Compared with normal tissues mean T-lymphocytes values were elevated in both involved lymph nodes and uninvolved spleens from Hodgkin's patients. Lymphocytes bearing C3 receptors were correspondingly reduced in these tissues. Involved spleen T-cell values fell within the normal range. In normal tissues the sum of lymphocytes with surface immunoglobulin and sheep erythrocyte receptors fell in the range 89-108%. In six biopsies of Hodgkin's tissue the sum was outside the normal range (121-142%). This observation is compatible with surface immunoglobulin-coated T cells. Surface marker characteristics and intracellular immunoglobulin studies of small lymphocytes, lymphoblasts and Hodgkin's cells suggested that the neoplastic cells were of B lymphocyte origin.


Smooth muscle antibodies in Mycoplasma pneumoniae infection. Paired sera from forty-five cases of Mycoplasma pneumoniae (MP) infection associated with acute lower respiratory tract illness were examined by immunofluorescence for antibodies to smooth muscle. Twenty-five (56%) of these cases had smooth muscle antibodies (SMA) of IgM class. A significant (greater than or equal to 4-fold) increase in titre of these antibodies was demonstrated in fifteen of thirty-five patients with a significant rise in titre of MP antibodies. SMA of IgG class occurred in eleven of forty-five cases (24%), but a 4-fold rise in antibody titre was found only in two cases. Three of forty-five sera (7%) from healthy donors contained SMA of IgM class and eight sera (18%) SMA of IgG class. MP antigen did not absorb SMA. Liver tests were performed in twenty-nine patients. In eighteen patients SGPT values were moderately or slightly elevated. There was no correlation between the occurrence of increased levels of transaminases and the presence of SMA in serum. In a patient with active chronic hepatitis, who had had a high titre of SMA exclusively of IgG class for 2 years, SMA of IgM class appeared transiently in association with an acute respiratory illness due to MP.


Antibody-dependent direct cytotoxicity of human lymphocytes. I. Studies on peripheral blood lymphocytes and sera of patients with systemic lupus erythematosus. Antibody-dependent direct cytotoxicity (ADDC) is generally believed to be unrelated to T-cell function in experimental animals. The role of ADDC in humans and its clinical usefulesss was evaluated in patients with systemic lupus erythematosus (SLE) and normal controls. Peripheral blood lymphocytes from patients with active SLE were unable to lyse antibody-coated target cells in vitro to the same degree as lymphocytes from patients with inactive SLE and controls. Sera from patients with active SLE suppressed ADDC by lymphocytes derived from normal controls and this abnormality was not corrected by overnight incubation or by extensive washing of lymphocyte preparations. Although there was poor correlation between ADDC and the proportions of B cells and null cells in effector lymphocyte populations from SLE patients and controls, it is concluded that this assay provides another means of determining immune competence in man.


Antibody-dependent direct cytotoxicity of human lymphocytes. II. Studies on peripheral blood lymphocytes, synovial fluid cells and sera from patients with rheumatoid arthritis. Antibody-dependent direct cytotoxicity (ADDC) by human lymphocytes was evaluated in patients with rheumatoid arthritis and normal controls. Purified peripheral blood and synovial fluid lymphocytes mediated normal ADDC when compared to control subjects. No correlation could be obtained between the percentages of T, B and null cells in effector cell populations and the degree of 51Cr released from target cells. Sera from 50% of patients with rheumatoid arthritis inhibited ADDC by normal lymphocytes; the degree of inhibition did not correlate with the titre of IgM rheumatoid factor. The pathogenic implications of these findings are discussed.


Inhibitory effect of sheep erythrocyte fragments on rosette formation of human T lymphocytes with sheep red blood cells. The effect of sheep red blood cells (SRBC) fragments on rosette formation of human peripheral T lymphocytes with SRBC was evaluated on the active and total T-rosette tests. The rosetting capacity of active rosette-forming cells was selectively and nearly completely inhibited by the pretreatment of lymphocytes with SRBC fragments. The decrease in total rosettes by blocking with SRBC fragments was almost parallel to that of active rosettes. SRBC fragments had no inhibitory effect on the rosetting capacity of a lymphocyte population in which active rosette-forming cells were removed by gradient centrifugation. These results suggested that active rosette-forming cells in human T lymphocytes have the receptors of high affinity for SRBC and these receptors readily bind SRBC fragments, resulting in block of rosette formation.


Multiple hereditary osteochondromata. Report of an early case. Observations on the early development of multiple osteochondromata in a 3-year-old girl suggest that aberrant growth of a peripheral segment of the growth plate, with extension primarily toward the metaphysis, but towards the epiphysis, impairs development of adjacent areas of epiphyseal cartilage.


Gastrointestinal complications of aortic bypass surgery. Gastrointestinal bleeding following abdominal aortic bypass surgery is not uncommon, as approximately 20% of patients with abdominal aortic aneurysms have peptic ulcer disease. We have recently seen three patients who presented with gastrointestinal bleeding secondary to the complications of their surgery. The cause of the bleeding was aortoduodenal fistula, graft erosion into the sigmoid colon and ischaemic colitis respectively. The correct diagnosis was only considered in one patient, although in retrospect it should have been suggested in all three. Our experiences with these complications and their clinical and radiological presentation form the basis for this paper.


Release of colony-stimulating activity from the isolated perfused rat liver. 1. Colony-stimulating activity appeared in the perfusate of the isolated rat liver during perfusions with either whole rat blood, rat plasma or an artificial perfusate of Eagle's medium and albumin. 2. Dialysable inhibitors of colony formation were also released during perfusions. 3. Colony-stimulating activity in artificial perfusate could be enhanced by the addition of rat plasma in vitro. Concentrations of cycloheximide that inhibited albumin synthesis by the liver did not inhibit the release of colony-stimulating activity.


Cardiac tamponade complicating closure of a median sternotomy. A case of intraoperative cardiac tamponade manifested during closure of a median sternotomy is presented. We postulate that cardiac tamponade was caused by acute dilatation of the cardiac chambers as a result of intraoartic balloon pumping in a patient with aortic and mitral regurgitation. It has been shown experimentally that acute rises in ventricular end-diastolic pressure result in increased intrapericardial pressure and that if a certain point on the pericardial pressure-volume curve is reached, cardiac tamponade will occur. Sternotomy closure was accomplished easily as soon as the need for intra-aortic balloon pumping diminished.


[Ulcerative colitis and pregnancy. Surgical treatment of acute complications (author's transl)]. Ulcerative colitis occurs chiefly in women in the reproductive age group. The disease tends to exacerbate in affected patients with florid colitis at time of conception. Toxic complications and severe haemorrhage are indications for surgical intervention at the earliest possible time. Treatment of choice consists of multiple enterostomies. Two female patients could be saved with this method. Proctocolectomies were always unsuccessful.


Acute hemorrhage from right-colonic hemangiomas: report of a case. A rare case of severe rectal bleeding beginning in adult life is reported. A diffuse, cavernous hemangioma of the right transverse colon was visualized by superior mesenteric arteriography and the diagnosis was confirmed by peroperative colonoscopy carried out through a colotomy. The patient was treated by right hemicolectomy. The diagnostic value of selective abdominal arteriography in patients who have severe rectal bleeding is demonstrated.


Immunoreactive insulin in bile and pancreatic juice of rat. Immunoreactive insulin (IRI) was determined in the bile and pancreatic juice of rats. Recovery studies in which known amounts of insulin were added to samples of these fluids indicated that the IRI values corresponded closely to total recovery. The concentration of IRI in bile and pancreatic juice was greater than the concentration in arterial plasma and less than the concentration in hepatic portal venous plasma adjacent to the liver. Elevation of the circulating IRI concentration in fasting anaesthetized animals by intravenously administered glucose or insulin produced a concomitant increase in the concentration of IRI in bile and pancreatic juice. In fasting conscious animals there was a circadian variation in the concentration of IRI in bile and pancreatic juice similar to the variation of IRI in the circulation. The total amount of IRI in the bile and pancreatic juice of fasting conscious rats during 24 h was in excess of 4 mU/kg body weight. These studies demonstrate that considerable quantities of IRI are transferred from the circulation into the bile and pancreatic juice of rats.


Anorectal melanoma: report of two cases. We have described the clinicopathologic findings in two cases of anorectal melanoma, and extracted the salient features from the medical literature. The disease is rare. Melanoma arises from the anal squamous membrane and very often spreads upward through submucosal planes, producing secondary satelites in the rectum. Trauma from defecation, vast lymphatic and venous systems in the anorectal region, and high invasiveness of the tumor cells eviden;ly account for early distant metastases. Histologically, the neoplastic cells often mimic other cancers. Treatment is surgical, with dismal end results.


Interrelation between anterior pituitary weight and ceruloplasmin level in serum after estrogen administration in male rats. Male rats were injected 11 mg estradiol benzoate in microcrystalline water suspension i. m. twice a week for three weeks. A significant correlation was found between the weight of anterior pituitary and ceruloplasmin level in serum (r = 0.358; P less than 0.01).


Diurnal variation in the effect of melatonin on plasma and muscle free fatty acid levels in the pigeon. Pigeons maintained on standard diet and held under 12 h daily photo-period in a controlled environmental room, were given intravenous injections of melatonin. A low dose (1.25 mg/kg body weight) of melatonin when given in the middle of the scotophase, produced a significant increase in plasma FFA when estimated at 20 min and 90 min post-injection, whereas no significant change was seen with injections given in the middle of the photophase. No significant change in muscle FFA level was obtained either during the photophase or the scotophase when estimated at 90 min postinjection. With a higher dose (5 mg/kg body weight) of melatonin given in the scotophase, on the other hand, a significant increase in both plasma as well as muscle FFA levels was obtained at 90 min post-injection but there was no effect on plasma FFA at 20 min or 90 min post-injection in the photophase and at 20 min in the scotophase. It is concluded that melatonin has a lipid mobilizing action in the pigeon when administered during the scotophase.


Adrenocortical changes in rats during acute and chronic administration of delta-9-tetrahydrocannabinol. Acute intraperitoneal administration of delta-9-THC (at doses of 10 mg/kg and 50 mg/kg) to adult male albino rats showed characteristic stimulating effect on adrenocortical tissues with marked depletion of ascorbic acid, cholesterol containing neutral lipids and significant increase in the delta5-3 beta -hydroxysteroid dehydrogenase and glucose-6-phosphate dehydrogenase activities in the zona fasciculata and zona reticularis regions. Adrenal weights and the histometric measurements of the adrenocortical areas showed no significant changes. Chronic intraperitoneal administration of delta-9-THC (at doses of 10 mg/kg per day for 15 days) produced marked accumulation of cholesterol containing neutral lipids, with increase in ascorbic acid and delta5-3 beta-hydroxysteroid dehydrogenase and glucose 6-phosphate dehydrogenase activities in the fasciculata-reticularis regions. Adrenal weights and histometric measurements revealed marked hypertrophy of the adrenal glands.


Investigation of luteinizing-inducing effect of pinealectomy or thyroidectomy in different types of anovulatory syndromes in rats. The luteinizing-inducing effect of pinealectomy and of thyroidectomy was investigated in the constant estrous anovulatory (CEA) syndrome caused either by frontal hypothalamic deafferentation (FHD) or by neonatal androgen treatment (NA). Thyroidectomy failed to induce luteinization in both types of CEA syndrome. However, pinealectomy provoked the formation of corpora lutea in FHD, but not in NA induced CEA syndrome. It was concluded that the luteinizing-inducing effect of pinealectomy is a specific consequence of the removal of the pineal hormone-like principles. The differences in the mechanism of development of the CEA syndrome following FHD or NA might account for the failure of pinealectomy to elicite luteinization in the NA induced CEA rat.


Dynamics of biosynthesis of thyroglobulin subunits and their polymerization in rabbit thyroid slices in vitro. The dynamics of biosynthesis and aggregation of subunits into thyroglobulin (TG) was studied in vitro in rabbit thyroid slices incubated from 5 to 300 min in the presence of I-14C leucine. The incorporation of labelled amino acid was followed in total soluble and microsome-bound proteins, as well as its distribution in soluble protein fractions. The incorporation of lebelled amino acid into soluble and microsome-bound proteins increased with time of incubation. The analysis of individual soluble proteins indicated that the label was incorporated very early, not only into 3--8S, but also into a protein which corresponds to the 12S fraction. Maximal incorporation into 12S protein was achieved after 60 min of incubation and then the intensity of incorporation decreased, followed by an increase in the relative and absolute amount of TG. The appearance of 14C-leucine in the TG region was not observed before 30 min of incubation. The dynamics of incorporation of 14C-leucine into thyroid proteins indicates very rapid transformation of newly synthesized 12S subunits into TG. After two hours of incubation the newly synthesized TG already showed the same sedimentation properties as pre-formed rabbit TG or native rat 19S TG-125I thus demonstrating that maturation, similar to polymerization, is a very rapid process, i. e. the newly formed TG is quickly transformed into its mature form. Our previous studies and the results presented in this work suggest that 12S protein may also have a precursor character. The presence of 12S protein in thyroid extract is not always the result of dissociation of TG, but also a consequence of polymerization of the basic subunits of TG.


Prolonged oral administration of potassium upon aldosterone biosynthesis by rat glomerulosa tissue in vitro. Steroid production rate of adrenals derived from rats drinking a 0.3 M KC1 + 5% glucose solution for 7 days was compared to that of control rats drinking a 5% glucose solution in order to investigate the effect of potassium loading upon the early and late step of aldosterone biosynthesis. Following potassium loading the quartered adrenals produced more aldosterone but less corticosterone as compared to the control. Potassium loading resulted in an increased aldosterone production rate by capsular adrenals (z. glomerulosa) provided that the corticosterone concentration in the incubation medium was elevated either by incubating it together with the decapsulated adrenal or adding exogenous corticosterone (4--16 mug/ml) to the medium. The corticosterone to aldosterone converting capacity of capsular adrenals is markedly higher in the potassium-loaded rats than in the controls. In the first 15 minutes of incubation the corticosterone production rate of the two groups was equal, aldosterone production rate by capsular adrenals of potassium-loaded rats, being higher than that of control animals. Corticosterone output of capsular adrenals from potassium-loaded rats decreased more rapidly in course of the incubation than it did in control tissue. These results suggest that the increase in aldosterone secretion in vivo following potassium loading is due to the stimulation of conversion of corticosterone to aldosterone in the glomerulosa cells. However, the endogenous corticosterone production during the incubation of glomerulosa cells from pottasium-loaded rats decreases so rapidly that the cells are not capable of producing more aldosterone than the control ones in spite of activated 18-hydroxylase.


Changes in uptake of 3H-progesterone by female rat brain and pituitary from birth to sexual maturity. 3H-progesterone uptake by various parts of the brain, pituitary and skeletal muscle was compared in newborn, 5-, 10-, 15-, 20-, 25- and 50-day old female rats at 1 hr after a single intraperitoneal injection of 50 muCi/100g body weight. High uptake values in newborn animals and in those aged 5 days were found in all tissues investigated. A sharp decrease in accumulation was observed from birth and/or 5th day of life. The uptake by pituitary was persistantly higher than those by other tissues investigated. The ratio of the concentration of radioactivity between the tissues and the cerebellar cortex increased significantly only in the posterior hypothalamus of adult females (at the age of 50 days). In the pituitary the ratio tissue/cortex was already significantly higher in newborns. The high level of brain radioactivity in the youngest animals is probably a manifestation of high plasma concentrations of tritiated progesterone. The striking decrease in uptake of radioactivity by the brain and pituitary during the first two weeks of life most likely reflects the decrease in level of plasma radioactivity, as shown indirectly by the concomitant decrease in labelled progesterone uptake by the skeletal muscle. The increase in tissue/cortex ratio in the posterior hypothalamus with attainment of sexual maturity suggests the first appearance of specific binding capacity for progesterone which is assumed to be present in the pituitary already from birth.


Morphological changes of thymus and the thyroid gland after postnatal extirpation of pineal body. Extirpation of the pineal body of newborn rats was followed by the disorganization of thymic structure, follicular transformation and proliferation, viz. malignant transformation of its epithelial and connective tissue elements. Increase of connective tissue was also observed in the thyroid gland. The follicular cells contained crystal-like bodies and vesicles, whereas in the parafollicular cells the endoplasmic reticulum was increased and light granules appeared.


Changes of glucose metabolism after thyroidectomy in chequered water snake (Natrix piscator). Thyroidectomy reduced the rate of removal of glucose from the blood of the chequered water snake (Natrix piscator) maintained at 40 +/- 2.0 degrees C.


Melatonin in rat pineal, plasma, and urine: 24-hour rhythmicity and effect of chlorpromazine. The concentrations of melatonin in rat pineals and in samples of rat plasma and urine were measured by bioassay or radioimmunoassay. Melatonin excretion during the daily dark period (0.98 +/- 0.07 ng/12 h, by bioassay; 1.40 +/- 0.08 ng/12 h, by radioimmunoassay) was much greater than during the light period (0.18 +/- 0.04 ng/12 h, by bioassay; 0.40 +/- 0.04 ng/12 h by radioimmunoassay). (The radioimmunoassay is not completely specific when applied to materials extracted from utine.) Pineal glands and plasmas obtained from animals killed during the daily dark period also contained much more melatonin than samples from animals killed during the light period. Pretreatment with chlorpromazine slowed the disappearance of exogenous melatonin from rat plasma and markedly raised the concentrations of endogenous melatonin in both the pineal gland and the plasma.


Movement of histones in chromatin induced by shearing. Methylation of accessible DNA within chromatin by restriction modification methylases from Haemophilus influenzae was used to detect movement of histones along the DNA strand during chromatin manipulation. Methylation at different stages of chromatin preparation was followed by titration of the nucleoprotein with ploy(D-lysine), digestion of chromosomal proteins with pronase and analysis of the DNA-poly(D-lysine) complex in steep cesium chloride gradients. Comparison of the specific radioactivities in the peak fractions of the free DNA and the DNA-poly(D-lysine) complex, respectively, reveals that lateral movement of histones, relative to specific sites in the DNA marked by restriction methylases, occurs during manipulation and fragmentation of chromatin.


Phytochrome-mediated assembly of polyribosomes in etiolated bean leaves. Evidence for post-transciptional regulation of development. Light operating through phytochrome controls the proportion of total ribosomes present as polyribosomes in etiolated leaves of Phaseolus vulgaris. Similar responses but with slightly different time courses are elicited by brief red light treatment and by continuous far-red or white light. The increase in polyribosome proportions after red light treatment reaches a maximum within 2 h, after which the proportion steadily declines. Light treatment appears to lead to increased proportions of polyribosomes in higher size classes. This is most evident with continuous white light, but probably also occurs with red and far-red light. The increase in polyribosomes is due principally to cytoplasmic ribosomes although proportionately greater effects are observed in chloroplast ribosomes. Although cordycepin inhibits RNA synthesis by 85-90% within 3 h there is no depression of the light-mediated increase in polyribosome proportions when leaves are preincubated in the inhibitor for 3 h. The data therefore indicate that phytochrome is capable of controlling protein synthesis, and thus development, at a post-transciptional level.


[The influence of bilirubin on the mechanogram of isolated, spontaneously beating frog heart]. It was shown that bilirubin in its unconjugated form does not only harm the central nervous system. Clinical observations indicate that the toxic effect under certain conditions affects also the heart. In a simple experimental model the influence of free bilirubin on the activity of the heart was investigated. In 69 adult frogs the heart was isolated and exposed sequentially to solutions of different bilirubin concentrations while the heart actions were registered. There was no significant change in the activity of the heart under the influence of bilirubin although concentrations were reached which according to the results of other authors harm cell cultures or evoke in the newborn disturbances in various organs. As a possible explanation, the nonsusceptibility of the heart muscle to bilirubin, the short duration of exposure, and the different behavior of newborn and adult organs is discussed.


[Thrombolytic and pharmacodynamic properties of Aspergillus ochraceus protease]. The protease isolated from Aspergillus ochraceus exerts a strong thrombolytic action. In in vitro experiments and in tests with animals (mice, rats, dogs) it caused lysis of fibrinous clots. The thrombolytic effect of the protease is conditioned by its direct influence on the fibrin. Besides the thrombolytic action the protease, used within a definite ranges of doses, displays antithrombotic properties. The paper cites data on the acute toxicity (in mice, rats and rabbits) and on the influence of the protease on isolated organs and on the organism of the animals.


[Participation of the serotonin-reactive brain structure in certain forms of behavior in golden hamsters]. A viviacious play of young hamsters is shown to be accompanied by a drop of the serotonin level in the brain stem and the subsequent slumber - by its rise, while the corticosteroids content of the peripheral blood with the playful  behavior experiences no changes. Iprazid and 5-oxytryptophan inhibit the playful activity, while dioxyphenylalanina (DOPA) does not influence it. A similar depression of the serotonin level in the brain stem was also noted in an aggressive behavior and stress conditions arising when adult male-hamsters are grouped together. A conclusion is drawn to the effect that changes in the content of serotonin in the brain stem are not associated with the emotional colouration of the condition, but rather reflect the transition from the somnolence to a highly active behavior.


[Mathematical analysis of the n-cholinolytic effects in experiments with the isolated frog rectus abdominis muscle]. An inquiry into the nature of antagonism of alpha-tubocurarine and gentamycin to acetylcholine based on a mathematical analysis of cumulative curves obtained in experiments with an isolated musculus rectus abdominalis of the frog showed that the character of changes in the parameters of cumulative curves occuring under the effect of alpha-tubocurarine cannot be explained from the standpoint of Clark's (1929) "employment theory", but accords with deductions drawn from a more general theory of allosteric interaction of substances and receptors (Karlin--1967) in competitive antagonism. Variations in the parameters of cumulation curves occurring under the action of gentamycin may be regarded as a manifestation of non-competative to acetylcholine in conformity with both theories.


[Vestibular tests in studying the spectrum of action of psychotropic preparations]. As demonstrated by tests set up on rabbits, substances endowed with sedative properties empede habituation to an inadequate stimulation of the vestibular receptors, whereas stimulators facilitate it. When used in small doses imipramin acts as a psycho-stimulator and in large ones--as a tranquilizor.


[Potentiation of the action of reserpine on the frog as a characteristic effect of antidepressants]. Antidepressants-thymoanaleptics chlorimipramine, amitriptyline, imipramine, melitracen potenciated the inhibitory effect of reserpine in a frog (supression of the turning over from the back reflex) stronger than did antidepressants with less marked thymoanaleptic action (Lu-5-003, azaphen, quipazine, desipramine) and also neuroleptics (promazine, stelazine, haloperidol) cholinolytics (atropine and benectyzine) and stimulator--phenamine. The potentiating action of tertiary amines--imipramine, amitriptyline protixen was stronger than that of the corresponding secondary amines (desipramine, norprotixen, nortriptyline). Phenothiazines with the chlorine atom in the second position (chlorprotixen and chlorpromazine) potentiated the effect of reserpine less intensively than did their chlorine-free analogues (protixen, promazine).


The role of the macrophage in genetic control of the immune response. The ability of an animal to mount an immune response is controlled by a number of autosomal dominant immune response (Ir) genes that are linked to the major histocompatibility complex of the species. In the guinea pig, alloantiserums raised by cross-immunization of inbred strain 2 and strain 13 animals specifically inhibited the in vitro proliferative responses of (2 X 13) F1 lymphocytes to those antigens the response to which is controlled by Ir genes linked to the alloantigens against which the serums are directed. A genetic analysis indicated that the inhibitory activity of the alloantiserums was directed against the alloantigens rather than the products of specific Ir genes. The interaction of antigen-pulsed macrophages with immune T lymphocytes is also mediated by the 2/13 alloantigens and alloantiserums are capable of inhibiting macrophage-T lymphocyte interaction. Studies involving combinations of macrophages and lymphocytes that differed at alloantigens or Ir gene products or both raised the possibility of the expression of the Ir gene product in the macrophage.


The role of suppressor cells in the pathogenesis of common variable hypogammaglobulinemia and the immunodeficiency associated with myeloma. The role of suppressor cells in the pathogenesis of immunodeficiency was analyzed using a technique that permits study of the differentiation of B lymphocytes into immunoglobulin-synthesizing plasma cells. Lymphocytes from normals synthesized 4,910 ng of IgM, 1,270 ng of IgA, and 1,625 ng of IgG per 2 X 10(6) cells when cultured for 7 days in the presence of pokeweed mitogen. In contrast the lymphocytes from patients with common variable hypogammaglobulinemia did not synthesize significant quantities of immunoglobulin. When lymphocytes from 9 of 13 patients with common variable hypogammaglobulinemia studied were cocultured with normal lymphocytes, the synthesis of immunoglobulin by the normal lymphocytes was depressed by 75-100%. A comparable suppression of immunoglobulin synthesis by normal lymphocytes was observed when they were cocultured with T cells from hypogammaglobulinemic patients. These studies suggest that in some patients the disease common variable hypogammaglobulinemia may not be due to an intrinsic defect of B cells alone but may be cuased or perpetuated by an abnormality of regulatory T cells that act to suppress B-cell maturation and antibody production. Peripheral blood lymphocytes from myeloma patients also had a drastically reduced capacity to produce polyclonal immunoglobulins. Three of 6 myeloma patients tested had circulating mononuclear cells that suppressed immunoglobulin production by cocultured normal lymphocytes. Purified T cells from myeloma patients did not mediate this suppressor effect. These observations suggest that one mechanism for the humoral immune deficiency observed in myeloma patients is a block of polyclonal B-cell maturation by suppressor cells.


Effect of serum proteinase inhibitors on the fertilizing capacity of rabbit spermatozoa. The in vivo effect of purified human alpha1-antitrypsin and inter-alpha-trypsin inhibitor on the fertilizing ability of capacitated rabbit spermatozoa was investigated and compared with that of the trypsin-kallikrein inhibitor from bovine organs (Trasylol). Only 250 mug of alpha1-antitrypsin/5 X 10(4) sperm in 0.05 ml inhibited fertilization by more than 50%. Lower concentrations of alpha1-antitrypsin (50 mug/5 X 10(4) sperm in 0.05 mo) and inter-alpha-trypsin inhibitor in amounts of 250 mug/5 X 10(4) sperm and less had no significant effect on the fertilization rate. Comparative experiments with Trasylol in conentrations of 50 mug/5 X 10(4) sperm resulted in more than 50% fertilization inhibition, whereas 100 mug/5 X 10(4) sperm decreased the fertilization rate by approximately 90%. The differences in the antifertility effect of these acrosin inhibitors toward capacitated spermatozoa may be due to differences in molecular weight. The effective concentrations required for fertilization inhibition appear to be relatively high under the experimental conditions used.


[The mechanism of development of changes in the structure and histochemistry of a neuron]. The change of the neuron's functional state when the cells become unable to generate spike responses to orthodromic testing stimuli, was shown not to be reflected in the morphological pattern of motoneurons as revealed by means of Einarsson's RNA staining. High-frequency stimulation of the motoneurons generating neither postsynaptic nor spike potentials diminishes the neurons size and intensifies their staining. A strong activation of presynaptic endings followed by no active postsynaptic processes on the neuron's membrane was shown to be able to entail major morphological alterations.


[Passive and active elements of the potassium secretion system in the skin of frogs]. A method has been evolved for studying the potassium and sodium transport without labelled ions in the frog skin. The potassium secretion sharply increased after the nystatinum had been added to the solution near the skin surface. Suppression of the apical membrane sodium permeability with amiloride or triamteren did not alter the potassium secretion velocity. The nystatinum increased both the potassium and sodium permeability. The blocking agents for sodium channels became ineffective in the presence of nystatinum. During the nystatinum action on the apical membrane the work of Na/K pump does not change in the basal plasmatic membrane, while suppression of the pump with strophantinum abolishes the potassium secretion across the apical membrane. Vasopressin and aldosterone increase the potassium secretion. Therefore, the potassium secretion level depends on the apical membrane potassium permeability and is independent of the sodium transport across this membrane, it is due to potassium inflow into the cell with the aid of Na/K pump of the basal plasmatic membrane.


[Responses of the neuromuscular synapses of chronically desympathized frog muscles to tetanic nerve stimulation]. 7--14 days after unilateral section of the abdominal sympathetic trunk, the quantum content of the end plate potential (e.p.p.) in the m. sartorius of the frog decreased. The facilitation and fluctuations of the e.p.p. amplitude during train of stimuli (20, 50, 100/sec) became more obvious. Noradrenaline (5-10(-6) g/ml) increased the amplitude of a single e.p.p. and the facilitation during repetitive stimulation. The effect of noradrenaline in sympathectomized muscles was more obvious than in the control muscles. After the noradrenaline administration, the fourfold increase of calcium concentration did not change the amplitude of e.p.p.s or the time course of the facilitation. The mechanism of depolarization--secretion coupling in the nerve terminal seems to be impaired by the sympathectomy.


[The effect of cholinesterase inhibitors on the synaptic potentials of frog neuromuscular junctions]. Prostigmine and the organophosphorus inhibitor armine acting upon synaptic potentials in the frog cutaneous pectori nerve--muscle preparation, increased the m.e.p.p. and e.p.p. amplitudes, lengthened the rise and the half--decline time of the e.p.p. in a-proximately similar way. The effect of armine developed faster than that of prostigmine. The presynaptic effects of both anticholinesterases (increase of mepp's frequency and quantal content of epp's) developed much more slowly than their post-synaptic effects. After prostigmine pretreatment during 2 hrs, addition of armine caused lengthening of the time-course of e.p.p.'s. but the presynaptic effect was not altered. After armine pretreatment, prostigmine did not cause any further effect. Possible mechanisms for the presynaptic effects of the inhibitors are discussed.


[Morpho-physiologic characteristics of the fiber spectrum of a frog external eye muscle]. By means of histochemical and microelectrophysiological methods, fibers of the eye inferior rectus muscle of the frog were divided into types with phasic, tonic (two types), and transitional (two types) signs. Electric activity of the muscle in situ reveals presence of phasic, transitional, and tonic fibers in the given object (the tonic fibers being characterized by a wide range of the PSP parameters).


[The influence of synaptic processes on changes in neuronal structure]. Reactions of the spinal motoneurons to the afferent inflow were different in frogs: some neurons generated spike potentials during 10-min rhythmic stimulation (1/sec) of afferents, other neurons were switched on and off. The motoneurons and their synapses increased in size, their cytoplasmic matrix became lighter and ribosoms deteriorated. Antidromic activation did not entail modofocation of size or color of the cells. RNP granules were safe and local organellas degenerated. The role of synaptic processes in regulation of RNP metabolism and in intracellular reparation, was revealed.


Retinopathy in Pima Indians. Relationships to glucose level, duration of diabetes, age at diagnosis of diabetes, and age at examination in a population with a high prevalence of diabetes mellitus. The occurrence of retinopathy and its relationship to diabetes in 1,640 Pima Indians age 15 and over has been determined. Eighteen per cent of those with two-hour postload plasma glucose levels of equal to or greater than 200 mg./dl. had some evidence of retinopathy. Of those with retinopathy and diabetes, 7 per cent were found to have proliferative or neovascular changes, the remainder having microaneurysms and/or exudates. The frequency of retinopathy increased from 3 per cent among newly diagnosed diabetics to 47 per cent among those with diabetes of 10 or more years duration. No relationship was found with sex, age at diagnosis of diabetes, or age at time of examination when duration of diabetes was taken into account. The occurrence of retinopathy was confined largely to those who fell into the second or hyperglycemic component of the frequency distribution of plasma glucose levels in the population, indicating the significance of the bimodal glucose tolerance frequency distribution.


Coronary heart disease in the Pima Indians. Electrocardiographic findings and postmortem evidence of myocardial infarction in a population with a high prevalence of diabetes mellitus. The frequency of electrocardiographic evidence of coronary heart disease (CHD) and the rate of autopsy-proved myocardial infarction were determined in the Pima, a tribe of American Indians with a high prevalence of diabetes mellitus. The electrocardiograms of 701 Pimas, aged 40 years and over (85 per cent of the adult reservation population, 45 per cent of whom had diabetes) were read according to the Minnesota Code, and 120 postmortem examinations were reviewed for evidence of myocardial infarction. The frequency of CHD as evidenced by major Q-wave changes in the Pima (1.6/100) was about one-half that found in Tecumseh, Michigan (p less than 0.10). The relatively low rate of myocardial infarction at autopsy (15 per cent of males and 8 per cent of females aged 40 years and over) was consistent with the low prevalence of Q-wave changes. The subjects with diabetes had a higher rate of CHD than nondiabetics, both electrocardiographically and at postmortem examination, although the differences were not statistically significant. The low prevalence of CHD in the living Pima and the low rate of infarction at autopsy indicate that this tribe has a low frequency of CHD despite the extraordinarily high prevalence of diabetes mellitus.


Diabetes mellitus: incidence, prevalence, survivorship, and causes of death in Rochester, Minnesota, 1945-1970. The incidence and prevalence of diabetes mellitus in residents of Rochester, Minnesota, for 25 years (1945 to 1970) were determined from available medical records. The over-all incidence rate for diabetes is 133 new cases per 100,000 population per year (age-adjusted to 1970 U.S. white population). The rate increased with age for both men and women and was higher among men over 30 years of age. The average annual incidence rates per five-year period for juvenile-onset diabetes mellitus were low and variable and showed little change. Polyuria, polydipsia, glycosuria, lean habitus, loss of weight, and high levels of fasting hyperglycemia at initial diagnosis occurred more frequently in younger than in older patients. The peak incidence in 1960 through 1964 and the decrease in the following five years may be a reflection of the introduction of the AutoAnalyzer method for blood glucose in 1958. The average annual incidence rates for 1955 through 1959 and 1965 through 1969 were essentially the same. The over-all prevalence for diabetes mellitus is 1.6 per cent, with a higher rate among men than among women over 40 years of age; among school children the rate is 0.1 per cent. Survivorship in the diabetic population is lower than that in the general population. The leading cause of death was coronary heart disease, the death rate from it being higher than for the general population.


The immune response in cirrhotic rats. The induction of tolerance to orally administered protein antigens. Hepatic cirrhosis reduced the susceptibility of rats to the induction of tolerance by the oral administration of a protein antigen. Rats with portacaval shunt were rendered tolerant as readily as normal rats. The orally induced state of partial tolerance was shown to be dependent on thymus-dependent lymphocytes: B lymphocytes reacted normally to challenge when injected with T lymphocytes from normal rats. Several factors may contribute to the reduced responsiveness of the cirrhotic rats to the tolerance regime. First, the cirrhotic liver was shown to have a reduced capacity to separate immunogen from tolerogen. Second, because of the reduced phagocytic capacity of the liver, increased quantities of lipopolysaccharide, derived from intestinal microorganisms, enter the blood stream. These substances and products of hepatocyte necrosis have adjuvant activity and may therefore contribute to the changed state of responsiveness of rats with cirrhosis.


Fiberoptic endoscopy during laparotomy in the diagnosis of small intestinal bleeding. A patient with recurrent gastrointestinal bleeding caused by hereditary hemorrhagic telangiectasia localized to the small intestine is described. The diagnosis was made by combining two recognized investigational tools-fiberoptic endoscopy and laparotomy.


Mallory-Weiss syndrome. Characterization of 75 Mallory-weiss lacerations in 528 patients with upper gastrointestinal hemorrhage. Seventy-five Mallory-Weiss lacerations were visualized endoscopically in 58 of 528 patients evaluated acutely for upper gastrointestinal bleeding. The Mallory-Weiss mucosal laceration is more common than generally recognized, is usually associated with hiatal hernia and a prodrome of retching or vomiting, and the ingestion of alcohol or acetylsalicyclic acid, or both. The lacerations are most commonly gastric and are associated with other mucosal lesions which may in fact be the instigating cause of the retching and vomiting. Although blood loss may be considerable, 90% or more patients with this lesion can be managed nonsurgically with appropriate blood component replacement and occasional use of systemic pitressin. There was one fatality in this series of 58 patients with Mallory-Weiss lacerations.


Functional T cells in rabbits gut mucosal lymphocytes. Functional studies of Peyer's patch and gut mucosa lymphocytes were performed in mixed lymphocyte culture reaction (MLC), an in vitro measure of the capacity of T cells to recognize and respond to cell surface alloantigens. The data demonstrate the functional effectiveness of both these mucosal lymphocyte populations in one-way MLC, both as responding as well as stimulating cells. Both the responding and stimulating capacities of these cells in MLC were almost the same as observed for lymphocytes obtained from either spleen or thymus. We conclude that functional T cells exist in both rabbit gut lamina propria and Peyer's patches with approximately the same quantitative capacity to respond in the MLC.


T cell depletion in untreated adult coeliac disease. The proportional and absolute numbers of circulating thymus dependent lymphocytes (T cells) were reduced in untreated patients with coeliac disease but were normal after treatment with a gluten free diet. There was an inverse correlation between circulating T cell numbers and jejunal intraepithelial lymphocytes. This evidence suggests a possible role for T cells in the pathogenesis of coeliac disease and is a further example of disturbed cell mediated immunity in this condition.


[Diverticulosis--diverticulitis].

[Diverticulosis--diverticulitis]. Diverticulosis is found in approximately 5% of people living in western industrialized countries. 95% of these are without any symptoms and do not need any therapy. For prophylaxis of diverticulitis a diet containing non-digestable cellulose and swelling substances is recommended. Complications of diverticulosis like diverticulitis, perforation abscess, or stenosis can necessitate a resection of the sigmoid colon. This should be done before life-threatening complications like free perforation or massive bleeding occur.


Lymphocyte trapping. Differential effects of ATS and irradiation on trapping in lymph nodes and spleen. Increased sequestration (trapping) of lymphocytes occurs in lymphoid organs following antigenic challenge. The effects of irradiation and anti-thymocyte serum (ATS) upon lymphocyte trapping in lymph nodes and spleen were studied. Both the above agents diminished or abrogated trapping in the draining lymph nodes; these same agents resulted in enhanced trapping in the spleen. Suppression of lymph node trapping could be achieved with a low dose of ATS and with 850 R, but not 200 R. These results are interpreted as showing that lymphocyte trapping in lymph nodes is initiated primarily by an ATS- and irradiation-sensitive population, whereas in the spleen a cell population resistant to the above agents activates the lymphocyte trap. These data demonstrate that the cell populations and/or the mechanisms involved in lymphocyte trapping in lymph nodes and spleens are distinct.


Isolated mesenteric injury due to blunt abdominal trauma. Isolated injuries of the small bowel mesentery or mesocolon with subsequent bowel infarction due to blunt abdominal trauma are rare. Two cases are described: 1 involving the mesentery to the terminal ileum and 1 involving the transverse mesocolon and middle colic artery, both with bowel infarction. The modes of clinical presentation and management of patients with injuries to the mesentery, mesocolon and mesenteric vessels following blunt trauma are described.


Activation of purified human T cells by mitogens: diminished mitogen-induced deoxyribonucleic acid synthesis in human T cells compared with autologous peripheral blood lymphocytes. Human thymus-derived (T) cells were isolated from peripheral blood after rosette formation with neuraminidase-treated sheep erythrocytes (SRBC). After separation on Ficoll-hypaque, SRBC were removed from T cells by treatment with tris(hydroxymethyl)aminomethane-NH4Cl. Human T cells and autologous peripheral blood lymphocytes (PBL) were then incubated with phytohemagglutinin, concanavalin A, or pokeweed mitogen. Human T cells, in the absence of other cell types, responded with less deoxyribonucleic acid (DNA) synthesis (measured by uptake of [3H]thymidine) than equal numbers of autologous PBL. Further experimentation established that, compared with autologous PBL, the diminished capacity of human T cells to be activated by mitogens was due neither to differences in the mitogen dose-response relationship nor to the time of peak DNA synthesis of T cells or autologous PBL. Fragments or components of SRBC were not detected on human T cells, and treatment of the T cell-SRBC mixture with tris(hydroxymethyl)aminomethane-NH4Cl did not contribute to the results. Increased cell density or a period of preculture before addition of mitogen also did not influence the degree of decreased DNA synthesis in human T cells compared with the response of autologous PBL incubated with the same mitogens. When mixed with the cells remaining at the Ficoll-media interface, purified human T cells did not suppress the mitogenic response of this cell mixture to PHA or concanavalin A. The data indicated that human T cells, in the absence of other cell types, were activated by mitogens to a lesser degree than autologous PBL. Furthermore, T cells responded with DNA synthesis after direct T cell-mitogen interaction.


Suppressive effect of a chronic helminth infection, schistosomiasis mansoni, on the in vitro responses of spleen and lymph node cells to the T cell mitogens phytohemagglutinin and concanavalin A. Chronic murine schistosomiasis mansoni is associated with depressed cell-mediated immune responses to Schistosoma mansoni egg antigens. The present study has examined the possibility that factors develop during infection that are capable of altering the response of lymphocytes to stimuli other than specific schistosomal antigens. Egg production begins at 5 weeks, and 1 to 3 weeks later there is a moderate degree of unresponsiveness of lymph node and spleen cells to the mitogens concanavalin A and phytohemagglutinin. This was associated with an altered dose response curve to the mitogens similar to that observed in antigenic systems. Seven weeks after the initiation of antigenic stimulation (egg prodiction lymph node and spleen cells from chronically infected animals were profoundly unresponsive to all concentrations of concanavalin A and phytohemagglutinin tested. These investigations suggest that, in addition to possible blockade by serum antibody, other suppressive factors may be involved in the spontaneous modulation of immunopathology in chronic schistosomiasis. These are detectable 1 to 3 weeks after the onset of egg production and are prominent at 12 weeks. Such findings are consistent with, but do not prove, the existence of suppressor T cells in chronic schistosomiasis.


T-dependence of human B lymphocyte proliferative response to mitogens. Human peripheral blood and tonsil lymphocytes were fractionated on anti-Ig-coated Sephadex columns or by centrifugation after rosetting with native sheep erythrocytes. Both methods allowed the recovery of B and T-enriched populations the purity of which was checked by fluorescein-labelled anti-Ig serum, E and EAC rosette formation, and heterologous antisera specific for B or T lymphocytes. The proliferative response of T cells to PHA, Con A, PWM, and ALS was not found different from that of unfractionated cells, whereas no response of the B cells could be observed to these mitogens providing that no contaminating T cells were present. Addition of T lymphocytes to these unresponsive B cells allowed them to respond to phytomitogens, but not to ALS. X-irradiated T cells could, to some extent, replace the diving T lymphocytes; no T-replacing factor could be found in cell-free supernatants from T cells, whether or not they had been activated by mitrogens. This model of B-T cooperation appears useful for studying the differentiation and maturation of human B lymphocytes.


Differences between lymphoid cell populations of guinea pigs and mice as determined by the response to mitogens in vitro. The stimulation of guinea pig lymphocytes by phytohemagglutinin (PHA), concanavalin A (ConA), methanol-extracted residues of tubercle bacilli (MER), purified protein derivative of tubercle bacilli (PPD), dextran sulphate (DS) and E. coli lipopolysaccharide (LPS), was determined and compared with that of mouse lymphoid cells. The sources of lymphocytes tested were spleen, thymus, lymph nodes and bone marrow. The degree of activation of DNA synthesis by PHA and ConA was higher in guinea pig thymocytes and lymph node cells than in corresponding sources of mouse lymphocytes. The optimum degree of stimulation by PHA and ConA was approximately the same in guinea pig thymocytes, while ConA was by far a better stimulator than PHA for mouse thymocytes. All four B-cell mitogens tested (MER, DS, PPD and LPS) activated DNA synthesis in mouse lymphoid cells while only MER and DS were effective in guinea pig lymphocytes. A guinea pig spleen cell population depleted from B cells was not stimulated, neither by DS nor by MER, while it still responded to PHA and ConA. These results indicate that the proliferative response due to MER and DS occurs in the B-cell compartment. It is suggested that the differences between guinea pigs and mice with respect to their ability to develop a cell-mediated type immunity and to respond to T-independent antigens are related to differences in the relative proportions and degrees of maturation of T- and B-cell subpopulations, as reflected by the selective responsiveness to various mitogens.


Passive sensitization of tissue cells. V. The detection of chicken antibodies cytophilic for basophils and eosinophils. Basophil-cytophilic antibodies were domonstrated in sera of chickens injected with either sheep erythrocytes or avian tuberculin. Normal chicken basophils were passively sensitized in vitro, and two types of cytophilic antibodies were shown by rosette-formation: (a) heat-stable, mercaptoethanol-resistant antibodies, and (b) heat-labile, mercaptoethanol-sensitive antibodies. Serum from one of the chickens injected with sheep erythrocytes also passively sensitized normal, non-basophilic granulocytes which would appear from stained smears to be eosinophils. Cytophilic activity in the serum correlated with the rosette-forming reactions of circulating eosinophils and/or basophils from each of the sensitized donor birds.


Antibodies to nucleic acids in myasthenia gravis. An increased frequency of antibodies to native DNA, thymocytes, and striated muscle was found in patients with myasthenia gravis (MG). The prevalence of such antibodies lends considerable support to the concept of MG as an autoimmune disorder and militates in favor of major abnormalities in the thymic dependent immune system. There was no correlation between serum-blocking activity to acetylcholine receptor protein and antibodies to thymocytes.


An acquired color defect of the opponent-color system. An acquired unilateral color defect in a 22-year-old man has been investigated with standard clinical tests and by using techniques which, it is thought, test specifically for the sensitivity of the luminance and opponent-color systems. The spectral sensitivity of the defective left eye, using 1 degree 200 ms. test flashes on a white background, has a single broad peak at about 550 nm. and resembles the photopic luminosity curve; in contrast, the normal curve, measured in the same conditions, has three peaks at about 440, 520, and 600 nm. However, the subject's spectral sensitivity curve for detecting 20 Hz. flicker is quite normal and is similar to his curve for 200 ms. flashes. It has recently been proposed that the three peaks of the normal curve for 200 ms. flashes reflect the activity of the opponent-color system, whereas the single peak for flicker detection is related to the luminance system. The preceding observations may thus be interpreted in terms of a specific loss of the subject's opponent-color system and this would explain his poor color discrimination. His luminance system appears to be normal, and evidence is presented for the maintained function of red- and green-sensitive (but not blue-sensitive) cones. The spectral sensitivity of the subject's right eye is nearly normal, suggesting a precortical origin of the defect; however, there seems to be some abnormality in this eye, indicating a less developed form of the same defect.


An intact globe technique for electrophysiological studies on cornea. An intact globe method was developed to determine the characteristics of the cornea of the bullfrog, Rana catesbeiana. With this method the anterior chamber could be perfused and the transcorneal potential difference (PD) and electrical resistance determined. It was found for the endothelium plus stroma (epithelium scraped) that the PD was essentially zero and the electrical resistance was only a small fraction of that of the intact cornea. Elevation of K+ or decrease in Cl- concentration in the anterior chamber produced in intact corneas a large and rapid change in PD while with the epithelium scraped (stroma and endothelium intact) these elevations produced a negligible change in PD. It is concluded that ions can rapidly move across the endothelium and stroma of the cornea.


Similarity in properties and mapping of three Rec mutants of Haemophilus influenzae. Three Rec- mutants of Haemophilus influenzae have been studied with respect to their transformability, ultraviolet and mitomycin C sensitivities, spontaneous and ultraviolet-induced deoxyribonucleic acid breakdown, inducibility of lysogens, and the linkage of the three mutations to a streptomycin resistance marker. The data indicate that the three mutations cause the same phenotypic changes, and that they are all on the same gene. Transformability of the mutants is different when two different media are used for competence development, although transformability with the two competence methods is not different in a Rec- strain that is mutant at another gene.


Ultrastructural characterization of capsulated Haemophilus influenzae type b and two spontaneous nontypable mutants. Capsulated Haemophilus influenzae type b and two spontaneous mutants (classes I and II variants) were characterized by transmission and scanning electron microscopy. When cells were treated with type b-specific antiserum prior to manipulations for electron microscopy, sectioned capsulated cells had electron-dense, fibrous capsular antigen-antibody complexes around them. In negatively stained preparations, the complexes appeared as electron-transparent zones surrounding cells. In contrast, only residual electron-dense, extracellular material was seen in sectioned, untreated, capsulated cells, and electron-dense "bridges" connected adjacent cells in negatively stained preparations. No extracellular capsular material was seen around the class I and II variants. Characteristic electron-translucent regions were always observed within the cytosol of the class I cells, both in thin sections and by negative staining. These areas were located adjacent to the cell envelope separating the plasma membrane from the dense cytoplasmic matrix. At times, electron-dense, thread-like material extended from the dense cytoplasmic matrix to the plasma membrane. No such regions were seen in the capsulated and class II cells. Class I cells fixed with methanol or suspended in NaCl or phosphate-buffered saline prior to treatment with fluorescein-tagged type b-specific antiserum (FTA reagent) exhibited, by immunofluorescence, patches of capsular antigen along their sides. However, when fixed with glutaraldehyde or OsO4 or suspended in tris-(hydroxymethyl)aminomethane plus Ca2+ buffer prior to treatment with FTA reagent, no patches of capsular antigen were seen. Subsequent exposure of the latter cells to methanol followed by treatment with FTA reagent resulted in the reappearance of the patches of capsular antigen. Thus, in the class I variant the capsular antigen is unlikely to be surface located. Scanning electron microscopy revealed that class I and II variant cells within undisturbed colonies were regularly aligned side-by-side, whereas cells within colonies of the capsulated strain were randomly distributed.


Synthesis of envelope polypeptides by Haemophilus influenzae during development of competence for genetic transformation. Six polypeptides with apparent molecular weights of 95,000, 90,000, 80,000, 67,000, 64,000, and 43,000 were found to be characteristic of the cell envelopes of competent Haemophilus influenzae, and were synthesized entirely during the period of competence development. Two polypeptides with apparent molecular weights of 58,500 and 40,500 were synthesized during growth as well as during competence development, but were only associated with the envelope fraction of cells that had developed competence. The kinetics of synthesis of the competence-related envelope polypeptides showed a lag period of approximately 20 min. The observation of this lag period raises the question as to whether some of these competence-related polypeptides might be involved in the process of deoxyribonucleic acid uptake, since the development of this property also exhibits a sigmoid time course during competence development.


The mechanism of action of ethanolamine ammonia-lyase, an adenosylcobalamin-dependent enzyme. The source of the third methyl hydrogen in the 5'-deoxyadenosine generated from the cofactor during catalysis. Ethanolamine ammonia-lyase is an adenosylcobalamin-dependent enzyme which catalyzes the conversion of ethanolamine and propanolamine to ammonia and the corresponding aldehydes. A mechanism has been proposed for this and other adenosylcobalamin-dependent reactions which involves cleavage of the carbon-cobalt bond of the cofactor followed by abstraction of a substrate hydrogen atom by the adenosyl fragment to form 5'-deoxyadenosine. In support of this proposal, a previous study demonstrated that the deamination of propanolamine by ethanolamine ammonia-lyase is accompanied by the reversible cleavage of the carbon-cobalt bond of the cofactor, with the production of 5'-deoxyadenosine (Babior, B.M., Carty, T.J., and Abeles, R.H. (1974) J. Biol. Chem. 249, 1689-1695). The present study is concerned with the origin of the third hydrogen atom on the methyl group of the 5'-deoxyadenosine produced in that reaction. The 5'-deoxyadenosine isolated from an incubation mixture initially containing enzyme, [5',5'-D2]adenosylcobalamin, and [1,1-D2]propanolamine was chemically degraded so that the 4' and 5' carbon atoms were, respectively, converted to the carbonyl and methyl carbons of acetaldehyde. Analysis of the p-nitrophenylhydrazone of the acetaldehyde by gas-liquid chromatography-mass spectroscopy revealed 3 deuterium atoms/molecule, indicating that two of the methyl hydrogens originated from adenosylcobalamin and the third was donated by substrate. This observation provides further support for the participation of 5'-deoxyadenosine in the mechanism of adenosylcobalamin-dependent reactions.


DNA synthesis and turnover in the bullfrog tadpole during metamorphosis. 125I-labeled deoxyuridine (IdUrd) has been used to estimate the turnover of DNA in liver, tail, and hind limb during spontaneous and triiodothyronine-induced metamorphosis. It was found that the total amount of liver DNA remained constant and there was no significant loss of the label from the liver DNA, which would be expected if there was an increase in DNA turnover during metamorphosis. Also, the change in specific activity of liver DNA parallels that of tail DNA during spontaneous metamorphosis. These data suggest that metamorphic transitions in the tadpole liver do not involve significant changes in DNA turnover. It was observed that the incorporation of label into hind limb DNA showed a high variability among individual animals as compared to liver and tail tissue. The data presented suggest that the observed variability is not a random phenomenon but related directly to the rate at which animals will metamorphose.


Multidrug chemotherapy in pulmonary treatment of osteosarcoma. Forty-three patients with osteosarcoma were treated with amputation and adjuvant chemotherapy utilizing a four-drug combination of cyclophosphamide, vincristine, phenylalanine mustard, and adriamycin (CONPADRI-I regimen). Twenty-four patients (56 per cent) remained free of metastases twelve to sixty-one months after diagnosis. Ten of the twenty-four have been disease-free for more than three years. Another group of thirty patients was treated with amputation and a five-drug adjuvant chemotherapy program which included the administration of massive doses of methotrexate with citrovorum factor (COMPADRI-II regimen). Twenty of the thirty (67 per cent) remained free of metastases from twelve to twenty-six months after amputation (median, sixteen months). Two deaths related to methotrexate toxicity occurred. Late metastases developed in three patients (at sixteen, nineteen, and twenty-six months after operation) in the group treated with the COMPADRI-II regimen.


BCG osteomyelitis: experience in the Stockholm region over the years 1961-1974. Eighteen cases of bone and joint tuberculosis in children were diagnosed in the Stockholm region (about 1,500,000 population) over the period 1961-1974. BCG infection was verified by culture and identification of bacterial type in seven, all after 1968. The same origin can be presumed in most of the remaining eleven cases, in spite of the absence of bacterial verification. The increased frequency of complications after BCG vaccination may necessitate a revision of the vaccination programme. We recommend operative treatment, which has not led to any growth disturbances or impairment of joint function, although the lesions were invariably localised close to growth zones and joints.


Synaptic activity of frog retinal photoreceptors. A peroxidase uptake study. The uptake of horseradish peroxidase (HRP) into membranous structures, detectable by light and electron microscopy, is used here to monitor the synaptic activity of photoreceptors of isolated frog retinas maintained in the dark or under various illumination conditions. The major findings are: (a) Neurotransmission from photoreceptor terminals seems to involve the same types of endocytic membrane-retrieval processes that occur at other nerve terminals. Presumably, the endocytic processes compensate for exocytic events associated with neurotransmission. The retrieved membrane is "recycled" to form vesicles. Some of these accumulate near the synaptic ribbons, perhaps indicating reutilization for exocytosis. On the other hand, some retrieved membrane evidently is degraded via multivesicular bodies that appear to undergo "retrograde" transport from the receptor synapses to the myoid regions. (b) Photoreceptor terminals take up much HRP in the dark. Steady illumination markedly decreases uptake by rods. Uptake by cones is notably reduced only at illumination intensities higher than those that have maximal effects on rods. (c) The decrease in rod HRP uptake with light is reversible when retinas are allowed to adapt to the dark, if the light exposures used were at intensities that bleach very little visual pigment. Such "recovery" is not observed after light exposures that bleach a considerable amount of visual pigment. The cones recover their dark levels of HRP uptake even after light exposures that bleach considerable amounts of visual pigment. The changes in HRP uptake that we observe parallel expectations for photoreceptor synaptic neurotransmission derived from indirect physiological evidence.


[Intracystic hemorrhages of pancreatic pseudocysts]. The authors report ten personal cases and review the literature on 99 other cases, and study primary intra-cystic hemorrhage or post-operative hemorrhage, occurring during the course of pancreatic pseudo-cysts. These hemorrhagic pseudo-cysts are very often associated with chronic pancreatitis; they may rupture into a hollow viscus, the peritoneal cavity or into Wirsung's duct. Arteriography and duodenoscopy with retrograde pancreatography, should permit one to reduce the frequency of these latent, undiagnosed forms until surgical operation has been carried out. Even more dangerous, post-operative bleeding often represents a recurrence of primary intra-cystic hemorrhage which has been undiagnosed or neglected. Surgical treatment should give a greater place to operations of excision which simultaneously treat false cyst, the responsible vascular lesion and the chronic casual pancreatitis.


The effects of epinephrine infusion in patients with von Willebrand's disease. Epinephrine infusion causes variable increases in the components of the Factor VIII (antihemophilic factor) complex in patients with von Willebrand's disease. The increase in antihemophilic factor procoagulant activity was greater than that of Factor VIII-related antigen and von Willebrand factor activity in two patients with von Willebrand's disease. Similar increases in the three individual factors were demonstrated in two other patients. A 4-10-fold increase in Factor VIII-related properties was identified in each of these individuals after infusion. One patient has been studied with very severe von Willebrand's disease; none of the Factor VIII-related properties increased despite two infusions of epinephrine. Bleeding times were normalized or remained normal in the two patients whose von Willebrand factor activity was greater than 25 U/100 ml. It remained prolonged in those three patients whose von Willebrand factor activity levels remained below that concentration. The increase in procoagulant activity was transient in all patients and t 1/2 values were estimated to be between 0.8 and 3.4 h.


Suppression of B-cell differentiation by leukocytes from hypogammaglobulinemic patients. Leukocytes from patients with several forms of immunodeficiency characterized by apparently differing defects in B-lymphocyte maturation produced few or no plasmacytoid cells in vitro, and were capable of suppressing the generation of plasma cells in co-culture with cells of normal persons, in the presence of pokeweed mitogen. Such inhibition was commonly observed in cultures which included cells from patients with primary immunoglobulin deficiency, but was not seen to a significant degree in identical co-cultures of cells from normals. The suppression observed was not dependent upon mixed leukocyte culture reactivity. Both sheep erythrocyte-rosetting lymphocytes and adherent cells appeared to participate in these effects in some patients. In one patient with common variable immunodeficiency, but not in several others, removal of suppressing cells permitted the patient's remaining cells to differentiate into plasma cells in vitro. Because of the diverse syndromes in which suppression was observed, it is likely that, in at least some hypogammaglobulinemic patients, the suppression is secondary to the disease process rather than being the primary pathogenic mechanism.


Heterogeneity of lymphocyte subpopulations in severe combined immunodeficiency. Evidence against a stem cell defect. Surface markers typical of T and B lymphocytes were present on varying proportions of peripheral blood lymphocytes from three infants with severe combined immunodeficiency disease. Despite this, functions mediated by T and B cells were either absent or very minimal in all three, including cell-mediated responses in vivo; the in vitro proliferative response to mitogens, allogeneic cells, or antigens; effector cell function in lymphocyte-antibody lymphocytolytic interaction assays; and in vitro synthesis of IgG, IgA, and IgM. In contrast, mononuclear cells from one of the infants were tested and found capable of lysing both human and chicken antibody-coated erythrocyte targets normally. Co-cultivation experiments with unrelated normal control lymphocytes failed to demonstrate suppressor cell activity for immunoglobulin synthesis in these infants. Augmentations of immunoglobulin production from 310 to 560% over that expected on the basis of individual culture data were noted in co-cultures of one of the infants' cells with two different unrelated normal control cells. These findings suggest that that infant may have had a T helper cell defect or that his T cells were unable to produce soluble factors necessary for B cell differentiation. The finding of cells with differentiation markers characteristic of T and B lymphocytes in each of these patients, though in variable quantities, is further evidence for heterogeneity among patients with the clinical syndrome of severe combined immunodeficiency and argues against the concept that their immunodeficiency was due to a stem cell defect.


Radioligand receptor assay for 25-hydroxyvitamin D2/D3 and 1 alpha, 25-dihydroxyvitamin D2/D3. A competitive protein binding assay for measurement of the plasma concentration of 1 alpha, 25-dihydroxyvitamin D3 [1alpha, 25-(OH)2D3] has been extended to include the immediate precursor of this hormone, 25-hydroxyvitamin D3 (25-OHD3). In addition, the assay system is capable of measuring the two metabolic products of ergocalciferol, namely. 25-hydroxyvitamin D2 (25-OHD2) and 1alpha, 25-dihydroxyvitamin D2 [1alpha, 25-(OH)2D2]. The target tissue assay system consists of a high affinity cytosol receptor protein that binds the vitamin D metabolites and a limited number of acceptor sites on the nuclear chromatin. By utilizing a series of chromatographic purification steps, a single plasma sample can be assayed for any of the four vitamin D metabolites either individually or combined. Therefore, the assay procedure allows for both the quantitative and qualitative assessment of the total active vitamin D level in a given plasma sample. To show that the binding assay was capable of measuring 1alpha, 25-(OH)2D2 as well as 1alpha, 25 (OH)2D3, two groups of rats were raised. One group, supplemented with vitamin D3, produced assayable material that represented 1alpha, 25-(OH)2D3. The other group, fed only vitamin D2 in the diet, yielded plasma containing only 1alpha, 25-(OH)2D2 as the hormonal form of the vitamin. The circulating concentrations of the two active sterols were nearly identical (15 ng/100 ml) in both groups, indicating that the competitive binding assay can be used to measure both hormonal forms in plasma. In a separate experiment, 1alpha, 25-(OH)2D2 was generated in an in vitro kidney homogenate system using 25-OHD2 as substrate. Comparison of this sterol with 1alpha, 25-(OH)2D3 in the assay system showed very similar binding curves; the D2 form was slightly less efficient (77%). Comparison of the respective 25-hydroxy forms (25-OHD2 vs. 25-OHD3) at concentrations 500-fold that of 1alpha, 25-(OH)2D3, again suggested that the binding of the D2 metabolite was slightly less efficient (71%). Finally, the assay was employed to measure the total active vitamin D metabolite pools in the plasma of normal subjects and patients with varying degrees of hypervitaminosis D. The normal plasma levels of 25-OHD and 1alpha, 25-(OH)2D measured in Tucson adults were 25-40 ng/ml and 2.1-4.5 ng/100 ml, respectively. Both sterols were predominately (greater than 90%) in the form of vitamin D3 metabolites in this environment. Typical cases of hypervitaminosis D exhibited approximately a 15-fold increase in the plasma 25-OHD concentration, and a dramatic changeover to virtually all metabolites existing in the form of D2 vitamins. In contrast, the circulating concentration of 1alpha, 25-(OH)2D was not substantially enhanced in vitamin D-intoxicated patients. We therefore conclude that hypervitaminosis D is not a result of abnormal plasma levels of 1alpha, 25-(OH)2D but may be cuased by an excessive circulating concentration of 25-OHD.


Clinical and laboratory studies into the pathogenesis of malacoplakia. Three cases of malacoplakia are described. Electron microscopic studies revealed intact bacteria or bacteria in varying states of degradation within phagolysosomes of the malacoplakic macrophages. Michaelis-Gutmann bodies arise within the phagolysosomes. These findings suggest that the bacteria incorporated within the phagolysosomes persist as dense amorphous aggregates which later become encrusted with calcium phosphate crystals to form the laminated Michaelis-Gutmann bodies. Possible explanations for the unusual macrophage response in malacoplakia are: (1) infection with an unusual strain of bacteria, (2) an immunological abnormality affecting intracellular killing of organisms, and (3) an abnormality affecting intracellular digestion of organisms. In considering each of the possibilities, we have been unable to detect any unusual strain of infecting organisms in association with malacoplakia, and in vitro studies have revealed normal T lymphocyte response to mitogen and normal monocyte bactericidal capacity. According to the history, each patient had reason to have a compromised immune status; in only one, however, was this demonstrated.


Defective T cell function in atopic dermatitis. The cellular immune system of 37 patients with atopic dermatitis (AD) was assessed by measuring peripheral blood T and B cells and the in vitro lymphocyte response to graded doses of phytohemagglutinin (PHA) (background and 6 concentrations of PHA from 100 to 1.6 mug). These were then correlated with clinical severity, ecosinophil counts, and serum IgE levels. The IgE levels (1,482 IU +/- 252 SEM), eosinophil counts (977 +/- 143), and absolute number of B cells (958 +/- 123) were significantly (p less than 0.05) higher than in age-matched controls (70 IU +/- 28, 182 +/- 79, and 480 +/- 60, respectively), and each significantly (p less than 0.05) correlated with the clinical severity. By contrast, percent B lymphocytes (20 +/- 1), percent (51 +/- 2) and total (2,357 +/- 217) T cells did not differ from controls. Eleven patients had low percent T cells (less than 40%); clinical and laboratory evaluation in these patients did not differ from the remaining 26. Lymphocytes from AD patients had higher background deoxyribonucleic acid (DNA) synthesis than controls (suggestive of increased number of B cells) and significantly depressed responses at the low PHA concentrations (6.3, 3.1, and 1.6 mug), which significantly correlated (p less than 0.05) inversely with IgE levels. These studies suggest a subtle defect in T lymphocyte function leading to increased B cells and increased IgE production.


Clinical conference: Severe obstructive lung disease in a 14-year-old girl with alpha-1 antitrypsin deficiency. Severe, largely irreversible obstructive lung disease, compatible with emphysema, was found in a 14-yr-old white girl who had been considered to have chronic asthma. She also presented a serum alpha-1 antitrypsin deficiency. Serum values for alpha-1 antitrypsin on two occassions were 95 and 105 mg/100 ml; Pi type was SZ. A family survey disclosed a 13-yr-old brother with the same pattern of alpha-1 antitrypsin  deficiency. His serum value was 122.5 mg/100 ml; Pi type was SZ. He was asymptomatic and showed minimal pulmonary function abnormalities.


Interaction between lymphocytes and inflammatory exudate cells. II. A proteolytic enzyme released by PMN as a possible mediator for enhancement of thymocyte response. Mouse polymorphonuclear leukocytes (PMN) were harvested from the peritoneal cavity stimulated with sodium caseinate. The cells were cultivated in vitro and the supernatant of these cultures (SUP) was tested for enhancing potency on DNA synthesis by syngeneic thymocytes stimulated with phytohemagglutinin (PHA). The enhancing potency of the SUP was markedly influenced by duration of the donor cultures of PMN, population density of the cultures, and protein concentrations in the medium, respectively. The enhancing factor in the SUP was found to be non-dialysable, heat-labile, and stable in the pH ranging between 3 and 9 but labile in the pH below 2 or above 10; its m.w. was approximately 19.000 when measured by gel-filtration on Sephadex G-75. The factor had a proteolytic activity on 3H-acetyl hemoglobin (3HHb) at neutral pH (7.2). Both the proteolytic activity and thymocyte-helping potency of the SUP were similarly abolished by adding protease inhibitor (Trasylol) in soluble form, or by passing through a column of the inhibitor insolubilized. It was thus assumed that the enhancing effect of PMN on thymocyte response was associated with a neutral protease released from the cells: it may be termed a lymphocyte-helping protease (LHP).


Identification of T and B cell subpopulations in human peripheral blood: electrophoretic mobility distributions associated with surface marker characteristics. The electrophoretic mobility distributions of human peripheral blood lymphocytes isolated on Ficoll-Hypaque gradients were characterized by laser Doppler spectroscopy. Three major subpopulations were spectrally resolved, due to differences in their mobility, when an electric field was applied to the scattering cuvette. The fastest component (centered at 2.35 mum/sec/V/cm for 25 degrees C, 0.28 M sucrose medium of 0.005 ionic strength) passed through nylon fiber columns and was identified as a T cell subpopulation. The slowest component (1.85 mum/sec/V/cm) which was further enriched by a one-step rosette procedure with sheep erythrocytes, reacted with activated-complement (C3) and antiserum to human immunoglobulins and was therefore identified as a B cell subpopulation. The intermediate component (centered at 2.15 mum/sec/V/cm) appears to be another T cellsubpopulation. Although cells of this component did not pass through nylon fiber columns, they did rosette with sheep erythrocytes. Furthermore, these cells did not appear to have surface immunoglobulins or complement receptors.


Mouse lymphotoxin.

Mouse lymphotoxin. The addition of PHA to C3H mouse spleen cells in tissue culture led to the production of lymphotoxin (LT). Cytotoxicity was assayed by addition of the culture fluids to syngeneic target cells labeled with tritiated thymidine; after an incubation period of 72 hr the amount of radioactivity released into the supernatant was measured. The LT activity in unfractionated culture fluids survived lyophilization, remained unchanged for many weeks at 4 degrees C, and progressively decreased on heating at 56 degrees C for periods from 1 to 9 hr. Based on the G-200 Sephadex distribution coefficients for several preparations, the m.w. of mouse lymphotoxin was about 41,000 daltons. Lymphotoxin from three different spleen cell production runs was recovered from isoelectric focusing columns in sharply focused peaks, the pH of which ranged from 4.4 to 4.8.


Studies on the maturation of immune responsiveness in the mouse. II. Role of the spleen. Experiments were designed to investigate the role of the spleen in the development of the murine immune system. By using mice splenectomized within 24 hr of birth, as well as mice with a hereditary, congenital absence of the spleen, the primary immune response to sheep erythrocytes was examined. The immunocompetence of lymph node cells from spleenless or control mice was assessed in vitro, in organ and in cell suspension cultures, and in vivo, by transfer into lethally irradiated syngeneic recipients followed by antigenic stimulation. The immunologic capacities of thymus and bone marrow cells were similarly tested by injection separately or in combination into irradiated syngeneic mice. Lymph node cells from spleenless animals appeared fully competent both in vitro and in transfer experiments. Neither neonatal splenectomy nor congenital absence of the spleen significantly reduced the capacity of bone marrow or thymus cells to participate in the immune response to sheep erythrocytes.


Relationship of CIF, LT, and PIF released in vitro by activated human lymphocytes. II. A further functional comparison of LT and PIF activities on HeLa and L-929 target cells. Lymphotoxin (LT), and proliferation inhibition factor (PIF) activities found in 5-day supernatants of mitogen-activated human lymphocytes (SAL) were further compared. In agreement with previous results, the activities could not be distinguished functionally. Quantitative differences in the amount of activity detected in the SAL could be accounted for on the basis of target cell differences, concentration of the lymphocyte effector molecules in the supernatant, and the parameter employed to assess cell function. Growth inhibitory activity detected at high supernatant dilutions was completely reversible, whereas the cytotoxic activity detected at low supernatant dilutions was irreversible. When the active medium was fractionated on DEAE, two peaks of inhibitory activity were detected. Depending upon the amount of activity and target cell, both peaks of activity were growth inhibitory or cytotoxic. Since both peaks of material affected HeLa and L-929 cells, the materials were not species specific. Thus, it appears that cloning inhibitor factor, LT, and PIF activities may actually be measures of the same stable materials found in 5-day activated lymphocyte supernatants.


The requirement for esterase activation in the anti-immunoglobulin-triggered movement of B lymphocytes. In this paper we bring evidence suggesting that there is activation of an esterase upon reaction of anti-immunoglobulin antibodies (anti-Ig) with murine B lymphocytes. B lymphocytes upon exposure to anti-Ig cap the ligand-receptor complexes and immediately afterward become briefly motile. It is this latter step which is inhibitable by exposure to di-isopropyl phosphofluoridate (DFP). Various experimental manipulations indicated that treatment with anti-Ig activates the cell for motility which, however, is not manifested until the temperature is raised to 37 degrees C. The cell incubated with anti-Ig at cold temperatures becomes susceptible to the effect of DFP, suggesting that the antibody-treated cells are activated up to but not beyond the DFP inhibitable step. Exposure of cells to DFP and removal of it before their treatment with anti-Ig does not affect the anti-Ig-induced response. Four lines of evidence indicate that the reduction of lymphocyte movement of DFP is due to the inhibition of an esterase activated by the combination of antibody and cell: 1) The inhibition by DFP is irreversible; once DFP has reacted it can be washed away and the antibody-treated cell is still inhibited. 2) The inhibition increases with time of contact of lymphocytes and DFP and with the concentration of DFP. 3) A very poorly phosphorylating phosphonate, phenyl ethyl pentylphosphonate is completely inactive under conditions where an excellent phosphorylating phosphonate, p nitrophenyl ethyl pentylphosphonate maximally inactivated the cells' movement. 4) The amino acid esters, tosyl L arginine methyl ester and benzoyl arginine methyl ester specifically prevent the inactivation by DFP. The last finding suggests that tosyl L arginine methyl ester and benzoyl L arginine methyl ester might be substrates for the putative antibody-induced lymphocyte esterase. Lymphocytes incubated with antibody in the cold for more than 30 min lose their ability to move when the temperature is raised, suggesting that there is a time-dependent deactivation of the cell.


Regulatory factors produced by lymphocytes. I. The occurrence of multiple alpha-lymphotoxins associated with ribonuclease activity. Supernatant fluids of mitogen-activated human tonsil lymphocytes contain large amounts of a factor toxic to mouse L cells. This substance, with a m.w. of 80,000 +/- 5,000 daltons, is called alpha-lymphotoxin (alpha-LT), to differentiate it from another toxin elaborated by mitogen activated human blood lymphocytes, called beta-lymphotoxin (beta-LT), which differs from alpha-LT in size (45,000 +/- 5,000 daltons), antigenicity, and stability. Further purification of alpha-LT by sequential phosphocellulose and DEAE-cellulose chromatography and polyacrylamide gel electrophoresis (PAGE) identifies a series of cytotoxins differing in ion exchange characteristics and electrophoretic mobilities. The three PAGE fractions (PAGE Ia, Ib and II), recovered in 2, 4.6, and 21% yield from the starting serum-free culture supernatant, represent purifications of 24-, 24- and 1851-fold, respectively. Each cytotoxic fraction has a ribonuclease activity. Comparison of RNase and mouse L cell cytotoxic activities of the three alpha-LT fractions shows that both activities for all three fractions have a similar temperature stability pattern and that both are similarly inhibited by DNA, single strand forms better than double strands, by glycerol in 5 to 20% concentration, and by protein denaturing reagents. These observations suggest, but do not prove, that mouse L cell toxicity and RNase activity are mediated by the same substance, which appears to occur in multiple or isozymic forms.


C1q (c1) receptor on human platelets: inhibition of collagen-induced platelet aggregation by C1q (C1) molecules. Hemolytically active human C1q incubated with EA before the addition of complement inhibited the immune hemolysis. On the contrary, heat-inactivated preparation (30 min 56 degrees C) was ineffective. Preincubation of EA with bovine collagen also resulted in a decreased hemolysis. When aggregation was measured by a turbidimetric method in citrated human platelet-rich plasma, it was found that hemolytically active human C1q (C1) alone does not induce platelet aggregation. However, in its presence the platelets failed to aggregate or exhibited a significantly reduced aggregation response to bovine collagen. The inhibition by C1q depended on the preincubation time with platelets. Heat treatment (30 min 56 degrees C) destroyed the inhibitory action of C1q (C1). The effect of C1q proved to be highly specific because different C1q preparations at their inhibitory doses in collagen-induced platelet aggregation did not influence the response to other aggregating agents (bovine thrombin, ADP, horse anti-human thymocyte globulin, goat anti-baboon platelet antiserum). The results prove that collagen and C1q are capable of binding to the same site(s); namely, to those of EA and human platelets; furthermore, they suggest the presence of a receptor for C1q (C1) on human platelets.


Suppressor cells in spleens from "nude" mice: their effect on the mitogenic response of B lymphocytes. A cell population recovered after velocity sedimentation fractionation of "nude" but not haired mouse spleen cells suppressed the response of spleen cells from both "nude" and haired mice to the B cell mitogen, LPS. The suppressor activity of these cells was abrogated by treatment with anti-theta antiserum and complement but not by treatment with the same antiserum preabsorbed with mouse brain. It is possible that these suppressor cells come from the pool of T cell precursors known to be present in "nude" bone marrow and spleen, and that they do not require thymic influence in order to perform the suppressor function.


Isolation and partial characterization of AgF-1:a rat lymphocyte membrane antigen. A genetically defined, serologically identified antigen of the rat lymphocyte membrane (AgF-1) has been isolated. Viable spleen and lymph node cells, prepared by Ficoll-Hypaque density centrifugation from Fischer rats, were radioiodinated with soluble lactoperoxidase. Extracts obtained with Nonidet P-40 were shown to contain numerous radiolabeled proteins including cell-surface globulin. AgF-1 was isolated from these extracts by precipitation with a highly specific alloantibody in conjunction with xenospecific anti-globulin antibody and polyethylene glycol (PEG). The use of PEG greatly increased the efficiency of the double antibody technique. The putative antigenic peak was eluted from sodium dodecyl sulfate polyacrylamide gels (SDS-PAGE) and specific antigenic activity was recovered. Removal of the SDS from these eluates was achieved by equilibration with urea and passage over an anion exchange resin. Renaturation, as evidenced by specific inhibition of complement-mediated cytotoxicity, occurred upon the removal of urea by dialysis. The m.w. of the purified antigen was estimated to be 35 to 40,000 daltons by SDS-PAGE and was unaffected by reduction with 2-mercaptoethanol. Amino acid composition was roughly similar to those reported for the major histocompatibility antigens of the rat and other species.


Generation of specific helper cells and suppressor cells in vitro for the IgE and IgG antibody responses. Normal splenic lymphocytes from BDF1 mice were cultured on ovalbumin (OA)-bearing syngeneic peritoneal adherent cells for 5 days and their subsequent helper function was tested by an adoptive transfer technique. Lymphocytes harvested from the culture were mixed with DNP-KLH-primed spleen cells and transferred into irradiated syngeneic mice followed by challenge with DNP-OA. The results showed that the cultured lymphocytes has helper function for both IgE and IgG anti-DNP antibody responses. Depletion of mast cells and T cells in the peritoneal adherent cell preparations did not affect the generation of helper cells in the culture. The helper function of the cultured lymphocytes was abolished by the treatment with anti-theta antiserum and complement and was specific for ovalbumin. The OA-specific helper T cells were generated in vitro by the culture of a T cell-rich fraction of normal spleen on T cell-depleted OA-bearing peritoneal cells. If the normal splenic lymphocytes or T cell-rich fraction were cultured with 10 mug/ml of OA in the absence of macrophages, cultured lymphocytes lacked helper function. The transfer of splenic lymphocytes or splenic T cells cultured with soluble OA to normal non-irradiated mice, however, suppressed both IgG and IgE antibody responses of the recipients to subsequent immunization with DNP-OA. The suppressor cells were sensitive to anti-theta antiserum and complement and their activity was specific for OA. The cultured cells transferred into normal mice did not suppress anti-hapten antibody response to DNP-KLH. Normal lymphocytes cultured on OA-bearing macrophages and had helper function in adoptive transfer experiments failed to suppress antibody response of non-irradiated recipients to DNP-OA. The results indicate that OA-bearing macrophages primed T cells and generated helper T cells, whereas the culture of normal lymphocytes with soluble OA in the absence of macrophages generated suppressor T cells.