Inhibition of human granulocyte function by methotrexate. Patients receiving cytotoxic drugs are at an increased risk of bacterial infection. Drug-induced leukopenia may be responsible for depression of host defenses; however, there is little information concerning the qualitative effects, if any, of cytotoxic agents on granulocyte antibacterial activity. Since methotrexate is now being used in massive doses in vivo, we investigated the effects of this drug on antibacterial and metabolic functions of normal polymorphonuclear leukocytes in vitro. Phagocytosis, quantitative protein iodination, and staphylococcal killing of normal polymorphonuclear leukocytes were found to decrease with exposure to increasing concentrations of methotrexate. The effects of methotrexate on these cell functions were rapid in onset and readily reversed by washing the cells, suggesting a locus of action on the cell membrane rather than at the level of nucleic acid synthesis. Exposure of cells to similar concentrations of folic acid or folinic acid produced no impairment of bacterial phagocytosis, suggesting that the observed effects are specific  for methotrexate. The concentrations of methotrexate that produced these impairments are readily achieved in vivo and may alter antibacterial defenses in patients receiving this therapy.


Altered toxicity of 5-fluorouracil following treatment with Corynebacterium parvum. Recent studies have demonstrated that systemic Corynebacterium parvum increases serum granulocyte-macrophage colony-stimulating factor and stimulates the proliferation of granulocyte-macrophage progenitor cells. It was hypothesized that more rapid cycling of granulocyte-macrophage progenitor cells would render the cells more sensitive to a cell cycle-specific chemotherapeutic agent. The colony-forming ability of bone marrow granulocyte-macrophage progenitor cells was assayed in vitro with soft agar cultures. C. parvum given before 5-fluorouracil in C57BL/6 mice increased the granulocyte-macrophage progenitor cell toxicity, the lymphopenic effect, and the lethality of 5-fluorouracil. When C. parvum was given after 5-fluorouracil, there was more rapid rebound of granulocyte counts to normal or supranormal levels.


Effects of human leukocyte conditioned medium on mouse haemopoietic progenitor cells. Medium conditioned by human peripheral blood leukocytes (HLCM) was studied for its in vitro effects on haemopoietic progenitor cells (CFU-s and CFU-c) present in mouse bone marrow. HLCM has poor colony stimulating activity in semi-solid cultures of mouse bone marrow cells, but invariably increases the number of colonies obtained in the presence of plateau levels of semi-purified colony stimulating factor (CSF). In liquid cultures, HLCM appears to contain a potent initiator of DNA synthesis in CFU-s, an activity which coincides with an increased CFU-s maintenance and causes a three- to four-fold increase in CFU-c number. It is apparent from this study that HLCM, in addition to stimulating colony formation in cultures of human bone marrow cells, has a profound in vitro effect on primitive haemopoietic progenitor cells of the mouse, which cannot be attributed to CSF.


A comparative study of physiological and structural changes at the myoneural junction in two species of frog after transection of the motor nerve. Structural and functional behaviour of motor end-plates after transection of the motor nerve has been studied in two species of frog: Rana esculenta and Rana temporaria. The physiological results show that in both species there is a transient cessation of spontaneous activity followed by a resumption of miniature end-plate potentials (min. e.p.p.s.) after denervation. The characteristics of these potentials (frequency, distribution of amplitudes, time-course) are similar in the two species. However, some differences have been observed: Firstly, the period of silence lasts for 2--4 days in the case of Rana temporaria whereas it is prolonged to about 15 days in Rana esculenta. Secondly, the resumption of min. e.p.p.s. is gradual and after the 10th day of denervation remains constant in Rana temporaria. It is inconstant independent of the period of denervation in Rana esculenta. The morphological results show that the Schwann cell is constantly in contact with the post-synaptic membrane after about 6 days of denervation in both species. It is suggested that either the Schwann cell is capable of functioning for a limited period of time in Rana esculenta or is activated to produce min. e.p.p.s. only in certain cases.


Influence of changes in composition of the cerebrospinal fluid on the secretory activity of the subcommissural organ in Rana esculenta. A quantitative histochemical and autoradiographic study by means of scanning cytophotometry. The effects of injection into the brain ventricle of noradrenaline dissolved in amphibian Ringer's solution, and of amphibian Ringer's solution alone on the secretory activity of the subcommissural organ (SCO) of Rana esculenta were studied. The parameters used were: the amount and mean concentration of secretory material within the SCO stained by aldehyde fuchsin (AF), the amount and mean concentration of radioactive material within the SCO labelled by 35S-cysteine, and the growth rate of the Reissner's or liquor fibre (LF). A significant influence on the amount of secretory material in the SCO was found to be exerted by the injection of amphibian Ringer's solution alone. This influence can be explained as an inhibition of the release of secretory material by the SCO. Injection of noradrenaline dissolved in amphibian Ringer's solution appeared to have a smaller inhibitory effect. Noradrenaline may be presumed to have only slightly stimulated the secretory activity of the SCO.


Scanning and transmission electron microscopy of the subfornical organ of the grass frog (Rana pipiens). The ventricular surface of the subfornical organ of the frog is made up of ependymal cells with numerous apical microvilli, occasional cytoplasmic protrusions and many vacuoles projecting into the lumen of the third ventricle. Between these cells dendrites of cerebrospinal fluid-contacting neurons reach the ventricle to terminate in bulbous enlargements. In addition, flask-shaped encephalo-chromaffin cells, containing granulated vesicles and aggregates of filaments in their cytoplasm, project into the cerebrospinal fluid. Surrounding the centrally located capillaries are enlarged dendrites and axons of heterogeneous morphology, some of which appear to originate within the subfornical organ, intermingled with dendrites and axons of normal structure. The glial cells in this region, especially the microglial cells, often contain large lipofuscin inclusions, suggestive of degeneration and subsequent phagocytosis of some of the enlarged dendrites and axons. The normally scarce neurosecretory peptidergic axons become more evident and form typical Herring bodies in stalk-transected animals. Neuronal perikarya of varying morphology are predominantly located peripheral to the region of enlarged dendrites and axons. Supraependymal macrophages are particularly numerous on the subfornical organ.


Ultrastructure of the age-involuted adult human thymus. Age involuted thymus tissue from a middle aged (33 years) and an old (63 years) man have been examined by electron microscopy and compared with thymus tissue from children. Biopsies had been taken during surgical correction of congenital heart defects. The fine structural architecture of cortex, medulla and connective tissue in the remaining lymphatic islands in the adult thymus investigated was not different to the thymus of children. We were surprised to find vigorous lymphocytopoiesis in the cortical regions and to recognize extended areas of medulla with a cellular composition which obviously provides the same microenvironment for T-cell maturation as the medulla of the non involuted thymus. Our findings are discussed in relation to the increasing arguments that the human thymus serves an immunological function throughout life.


Determination of 5-hydroxytryptophan in plasma by high-performance liquid chromatography and fluorometric detection after phthaldialdehyde reaction. We describe the sensitive and precise determination 5-hydroxytryptophan in plasma from patients treated with this compound. The method is based on solvent extraction of plasma, high-performance liquid chromatography and continuous fluorometric detection after reaction with phthaldialdehyde in strong acid. The combined time for chromatography and reaction is 25 min. The sensitivity of the method is adequate to quantitate 0.05 mumol of 5-hydroxytryptophan per liter, in 750 microliter of plasma. No interference by other hydroxyindole compounds was found.


The properties and clinical significance of some electrophoretically slow forms of alkaline phosphatase. Sera from 8 patients with a marked slow-moving alkaline phosphatase band on electrophoresis were investigated. Inhibitor studies and treatment with neuraminidase showed that all the patients had slow bands with alkaline phosphatase properties resembling those of the liver or bone isoenzyme. In no case did the slow band resemble the intestinal isoenzyme. Immunoelectrophoretic and molecular weight studies indicated that the slow band consisted of an IgG-alkaline phosphatase complex of molecular weight 540 000. Serum from a patient with the slow band was able to bind liver or bone, but not intestinal, alkaline phosphatase from other patients to form the slow band. Serum from patients with the slow band probably contains an abnormal IgG molecule which can bind alkaline phosphatase in the ratio 2:1. No clinical condition was common to all 8 patients although most of them had either intestinal or lung disease.


Bone disease in children with chronic renal failure: therapy with 1alpha-hydroxyvitamin D3. Results of 1alpha-hydroxyvitamin D3 therapy in twelve children with renal bone disease are described. Nine of the twelve children showed a good response to 0.05--0.08 microgram/kg/day. Hypercalcaemia was the only side effect and proved easy to manage because of the short half-life of 1alpha-OHD3.


Alpha-1-antitrypsin (Pi)-types in recurrent miscarriages. Chromosome analysis was carried out in a consecutive series of 57 couples with a history of recurrent miscarriage. All 114 subjects from these couples had normal karyotypes by conventional analysis. Pi typing was performed in 30 couples. The constellation of Pi heterozygosity in both partners occurred in two of the couples. The expected frequencies of the observed MS-MS and MS-MZ constellations occurring by chance would be 2.1 per 1,000 and 2.2 per 1,000 couples, respectively.


Familial syndrome of progressive cone dystrophy, degenerative liver disease, and endocrine dysfunction. III. Genetic studies. A syndrome of progressive cone dystrophy, endocrine dysfunctions and degenerative liver diseases has been observed in seven patients, six of whom belonged to one extensive kindred. Genetic analyses revealed a segregation ratio indicating autosomal recessive inheritance of the syndrome, and the kindred from which six of the seven patients originated was heavily inbred. Thus, the results of the segregation analyses as well as of the inbreeding analyses provide evidence that this previously unrecognized disorder is inherited as an autosomal recessive trait. Genetic marker analyses were conducted with respect to 22 marker systems, and linkage information was obtained with respect to 15 of them. No strong suggestion of linkage emerged from the analyses, but very close linkage could be excluded for several of genetic marker systems. Pedigree analysis was helpful in establishing the spectrum of clinical manifestations belonging to the syndrome proper. The data presently available suggest that elevated levels of creatine phosphokinase, which were found in all patients, may be useful in tracing heterozygotes for this disorder. This possibility will be further examined.


T and B lymphocytes in patients with chronic active hepatitis (CAH). Absolute numbers of T and B lymphocytes as well as active E rosette-forming cells were measured in twenty-seven patients with chronic active hepatitis (CAH), and in thirty control patients. In patients with CAH without cirrhosis, active E rosette-forming cells (a subpopulation of T lymphocytes considered to be actively involved in cell-mediated immune reactions) as well as lymphocytes with surface markers for IgA, IgM and IgG were increased. In patients with CAH and cirrhosis, total T lymphocytes were decreased. These results emphasize the significance of lymphocytes in CAH, and suggest the importance of monitoring T- and B-cell populations in patients with this disease.


Lymphocytotoxins in epileptic patients receiving phenytoin. The present study was undertaken to determine the prevalence and nature of lymphocytotoxic antibodies in patients receiving phenytoin. Of forty-six sera examined, lymphocytotoxins and antinuclear antibodies were detected in fourteen and eleven samples respectively. Antibodies to double-stranded RNA (poly-A-poly-U) were found in six out of thirty-eight sera studied. A highly significant correlation was found between the presence of lymphocytotoxins and antinuclear antibodies (chi2 = 9.72; P less than 0.01). Lymphocytotoxins were found to be of the IgM class, to have activity against T and B cells, and to be dependent on the presence of Ca and Mg ions for activity. These findings may have pathogenetic importance in the genesis of the altered immune state of patients reveiving phenytoin.


Stimulation of a human B-lymphocyte line by anti-immunoglobulin and its concanavalin A-induced suppression by a T-cell line. The effect of anti-immunoglobulin (anti-Ig) on cultures of established human B-cell lines was studied to develop models for the initiation and regulation of immune responses. DNA synthesis in one line, BM, measured by [125I]iododeoxyuridine (IUdR) incorporation, was stimulated by anti-Ig when the continuously proliferating cultures approached high cell density. One of four lines, BJAB, was very sensitive to inhibition of DNA synthesis by anti-Ig at all cell concentrations. Of four human T-cell lines tested for suppressor activity, the CEM line was found to be activated by concanavalin A (Con A) to inhibit the anti-Ig stimulation of BM cells. No cytotoxic activity was detected in Con A-treated CEM cells. Both the anti-Ig stimulation of BM and Con A-induced suppression by CEM occurred with cloned cell lines in long-term culture, in the absence of helper or accessory cells.


Blood and thyroid-infiltrating lymphocyte subclasses in juvenile autoimmune thyroiditis. We have studied the distribution of T and B lymphocytes in the blood and in the thyroid lymphocytic infiltrates (obtained by fine needle aspiration biopsy) in sixteen patients with juvenile autoimmune thyroiditis (JAIT). The same cell populations were also tested for cell-mediated immunity (CMI) to thyroid antigen in the leucocyte migration test (LMT). The relative and absolute numbers of blood T lymphocytes were normal (71-76%), as were the numbers of blood B lymphocytes (19%). The thyroid infiltrate contained 48% B lymphocytes, whereas only 40-44% of the infiltrating lymphocytes were T cells. Half of the JAIT patients showed a positive CMI to thyroid antigen with blood leucocytes, but when thyroid-infiltrating lymphocytes of these patients were tested in the LMT, they were negative. Thus, in contrast to what is generally assumed, we were unable to demonstrate T cell-dominated lymphocytic infiltrates or the accumulation of specifically sensitized T lymphocytes within the thyroid gland in autoimmune thyroiditis.


Subpopulations of human T lymphocytes. I. Studies in immunodeficient patients. T lymphocytes with receptors for IgM(Tmu) and IgG(Tgamma) were examined in thirty patients with primary immunodeficiency and autoimmune disorders. Six out of twenty-seven patients with primary immunodeficiency had a low proportion of Tmu cells when compared with normal controls. Eight out of twenty-seven patients with primary immunodeficiency had an increased proportion of Tgamma cells. Two out of twenty-seven patients had both a low proportion of Tmu cells and a high proportion of Tgamma cells. The patient studied with severe combined immunodeficiency had a low proportion of both Tmu and Tgamma cells. Patients with Bruton-type agammaglobulinaemia, common variable immunodeficiency, thymoma and immunodeficiency syndrome and selective IgA deficiency demonstrated heterogeneity with regard to alterations in T-cell subpopulations.


Influence of SRBC/lymphocyte ratio on T-cell rosettes in alcoholic liver disease and inflammatory bowel disease. Thymus-derived (T) rosette-forming cells were enumerated in patients with alcoholic liver disease and in patients with inflammatory bowel disease using variable sheep red blood cell (SRBC)/lymphocyte ratios. SRBC/lymphocyte ratios of 60:1 and 32:1 did not reveal significant differences from controls in Crohn's disease. The percentage, but not absolute count, of T cells was significantly reduced in alcoholic hepatitis at the 60:1 ratio. Both the percentage and absolute count of T cells were reduced in alcoholic hepatitis and Crohn's disease with the 8:1 ratio. No significant reduction in T cells was seen at any ratio in patients with compensated alcoholic cirrhosis or ulcerative colitis. Use of a SRBC/lymphocyte ratio of 8:1 indentifies T cells which demonstrate an avidity for SRBC. This avidity may be related to the density of SRBC receptors on the surface of T cells and/or the affinity of these receptor sites for SRBC. Use of the 8:1 ratio may provide a more sensitive means by which to monitor changes in T-cell rosettes in patients suspected of having an altered cellular immune state.


Identification of human mononuclear leucocyte populations by esterase staining. Histochemical staining for alpha-naphthyl (non-specific) esterase has been employed to define subpopulations of human peripheral blood lymphocytes and monocytes. To define optimal conditions for staining, various fixatives, incubation times and cell preparations were compared. The great majority of human blood lymphocytes were found to have discrete granules of reaction product. More than 80% of T lymphocytes separated by E rosetting are esterase-positive whereas non-T lymphocytes are esterase-negative. Lymphocytes transformed by polyclonal mitogens lose their esterase-staining granules, which suggests that immature T cells are esterase-negative. Most blood monocytes show a diffuse cytoplasmic esterase reaction product and are phagocytic. However, about 20% of diffusely stained cells are not phagocytic. When leucocytes are cultured for 24 to 48 hr, the total number of esterase-positive cells increases and the great majority are phagocytic. This is interpreted as maturation of precursors into mature esterase-positive phagocytic monocytes. When cultured for longer periods, some lose phagocytic capacity and acquire the characteristics of secretory cells. Esterase-staining of lymph node sections allowed the identification of T- and B-dependent areas as well as macrophages related to sinuses. The esterase-staining technique could play a useful role in clinical and experimental immunology.


Some immunological effects of penicillamine. Immunological effects of D- and D,L-penicillamine (PA) were studied in efforts to develop assays for synthetic D or D,L analogs and to contribute to the understanding of the mechanism(s) of action of D-PA in rheumatoid arthritis. At the highest doses tolerated by mice, D,L-PA did not significantly inhibit the development of haemagglutinating antibodies in vivo. In studies in vitro with T lymphocytes, D-PA at 1 mM concentration inhibited both concanavalin A- and phytohaemagglutinin-induced transformation as assayed by [3H]thymidine incorporation, but D-PA concentrations of 5 mM were required to inhibit concanavalin A-induced amino acid uptake. No effect of D-PA was observed either on the induction of cytotoxic T cells or on the attack of specifically sensitized T cells on target cells. It is of interest that D-PA at 1 mM concentration did inhibit lipopolysaccharide-induced transformation, which predominately stimulates B lymphocytes. The effects of PA on the induced transformation of T and B cells deserve further attention for studies with analogs of PA.


Human blood L lymphocytes in patients with active systemic lupus erythematosus, rheumatoid arthritis and scleroderma: a comparison with T and B cells. Human blood lymphocytes with high affinity Fc receptors for IgG will bind small aggregates of this immunoglobulin at 4 degrees C. These cells have been named L lymphocytes because of membrane-labile IgG determinants. L cells possess a profile of surface markers and functional characteristics which differ from T and B cells. Immunofluorescence methods have been employed to quantify L lymphocytes in subjects with connective tissue diseases and certain infections, and these values have been compared with those for T and B cells. The mean values of L lymphocytes in groups of patients with systemic lupus erythematosus, rheumatoid arthritis and scleroderma ranged between 14% and 18%; values similar to normals. Groups with acute pneumonia and tuberculosis, however, had significantly increased percentages of L lymphocytes. The absolute number of L cells was decreased in subjects with connective tissue diseases, as was the number of T and B cells. L lymphocytes in those with infections were not significantly decreased. Only L lymphocytes were depleted by immobilized antigen--antibody complexes, another characteristic which distinguishes them from T and B cells.


T- and B-lymphocyte subpopulations in pre-invasive and invasive carcinoma of the cervix. Lymphocyte subpopulations in patients with pre-invasive and invasive cervical carcinoma, other gynaecological malignancies and controls were studied. T lymphocytes were recognized by their ability to form spontaneous rosettes with sheep red blood cells (E rosettes). Two surface marker characteristics were used to detect B lymphocytes: the receptors for activated complement responsible for erythrocyte-antibody-complement (EAC) rosette formation, and surface membrane immunoglobulin (SMIg), which is readily stainable by immunofluorescence. There was a significant depression in T cells in association with invasive but not pre-invasive cervical carcinoma. The results for B cells varied according to the method used for their detection. EAC rosette-forming (EAC-RFC) were significantly raised in patients with invasive cancers but not in patients with pre-invasive cancer. SMIg-bearing cells were not significantly altered by the presence of malignant disease. The changes in E-RFC and EAC-RFC numbers were more marked in patients with extensive cancers. Possible functional implications of these findings are discussed.


Increase in peripheral blood 'null' cells in extrinsic bronchial asthma. Lymphocyte subpopulations were measured in the peripheral blood of fifteen patients with extrinsic bronchial asthma and twenty-seven normal control subjects. No significant differences in the absolute numbers of relative proportions of T, B or K lymphocytes were found. Antibody-dependent lymphocyte-mediated cytotoxicity (K-cell activity) was also found to be normal. The proportion and absolute number of 'null' cells were significantly increased in the asthmatic group: this population did not correlate with serum IgE levels, the eosinophil count or the duration of disease. The significance of these results in relation to suppressor T-cell control of IgE formation is discussed.


Effect of levamisole on autologous rosette-forming cells in nude mice. Levamisole depresses the number of autologous rosette-forming cells (ARFC) in the spleen of nude (congenitally athymic) mice. Intravenous administration of 2.5 mg/kg of levamisole produces maximal depression. This effect appears 15 h after injection and is transient, partially disappearing after 48 hr. Dexamisole is devoid of this depressing activity. Thymopoietin, a thymic hormone, is also shown to lower the level of autologous rosette formation. These results suggest a stereospecific interference of levamisole with the maturation of immature T-cell precursors in a manner resembling the action of thymic hormones.


The underfilled coronary artery: some pre- and post-operative observations on recipient arterial quality and left ventricular function after coronary artery surgery. In a series of 141 saphenous vein aortocoronary bypass grafts in 59 patients undergoing routine restudy of the coronary arteries within four weeks of operation, an overall graft occlusion rate of 12% was found. A trend towards a higher occlusion rate was seen in arteries that were small (19%), that had a limited run-off (15%), significant distal disease (24%) or severe atheroma at the graft site (22%). Statistically, these occlusion rates are not significantly different from the overall failure rate and do not give absolute contra-indications to grafting. A particular group of arteries which seemed underfilled in the pre-operative angiogram (1.5 mm or less in diameter but apparently healthy with smooth walls and supplying a larger area of myocardium than would be expected from their apparent size) was identified. These arteries showed a significant increase in diameter in the post-operative angiograms and had a low occlusion rate (1 of 15 grafts; 7%). When all grafts were patent, the mean post-operative left ventricular ejection fraction showed a significant increase over the pre-operative valve, but when one or more grafts were occluded there was no significant change.


[Malignant histiocytosis (author's transl)]. According to observations in six patients and published reports, malignant histiocytosis is characterized by premature multifocal proliferation of atypical histiocytes, especially in lymph-nodes, spleen, liver, bone marrow, and lung. The diagnosis can be confirmed by electron-microscopy, and enzyme as well as immunocytochemical tests. Fever, anaemia, leukopenia (with absolute reduction in T-lymphocytes), and jaundice are frequent. Immunoglobulins are normal or polyclonally increased. Malignant histiocytosis is more like monocyte leukaemia and histiocytosis X than neoplasms of the lymphatic system.


[Liver disease in homozygous alpha1-antitrypsin deficiency (author's transl)]. Among twelve patients with homozygous alpha1-antitrypsin deficiency (Pi-type Z), five cases of infantile liver disease were diagnosed. The course of the disease was extremely variable; only one patient died of liver cirrhosis at the age of fourteen. In four cases the clinical, biochemical and histological (2 cases) findings became normal over a follow-up period of one to fifteen years. The results of these observations demonstrate that in alpha1-antitrypsin deficiency even when associated with proven liver disease the prognosis need not be unfavorable.


Properties of isolated human alpha1-antitrypsins of Pi types M, S and Z. 1. alpha1-Antitrypsin contains a single thiol group partly blocked in native plasma and reactive after mild reduction. 2. Human alpha1-antitrypsins of Pi types F, M, S and Z have been isolated with native microheterogeneity using thiol-disulfide (SH-SS) interchange reactions utilizing the reactive thiol group. 3. The pI of the various microheterogeneous fractions are given for protein M. Stepwise desialylation of alpha1-antitrypsin indicates that the charge difference between the major fractions is one sialic acid residue between each. This is further supported by the pI changes obtained on substitution of the single thiol with positively or negatively charged compounds. 4. Desialyation of purified proteins from each Pi type converts the individual microheterogeneous fractions to one major fraction. The pI shift for the variants studied indicate a difference of plus or minus one or two charge units between protein M and the variants. 5. A difference of one sialic acid residue was obtained for proteins M and Z by the thiobarbituric assay, but stepwise removal of sialic acid with neuraminidase revealed almost identical stepwise change of pattern of both proteins indicating the same number of sialic acid residues. 6. Electrofocusing has been used to identify CNBr fragments from proteins M, S and Z. 7. An amino acid substitution has been found to be located in one of the eight CNBr fragments, glutamic acid in protein M is substituted by lysine in protein Z. 8. The average concentration of alpha1-antityprsin in plasma from healthy males was found to be 1.32 g/1.


Alpha-1-antitrypsin in umbilical cord serum: pi phenotypes and relationships with idiopathic respiratory distress syndrome. The concentrations and phenotypes of serum alpha-1-antitrypsin (alpha1AT) were determined in 650 newborn infants. The distribution of these 650 subjects among the various Pi phenotypes confirms the higher frequency reported for the PiS allele in Latin populations. Serum alpha1AT levels vary between one phenotype and the other. Besides, at birth, infants weighing more than 2,500 g have alpha1AT levels significantly higher (P less than 0.001) than infants weighing less than 2,500 g; this difference in serum alpha1AT concentrations is due to the low alpha1AT levels found in preterm infants. The significantly lower alpha1AT concentrations found in preterms is associated with a higher risk of developing IRDS and with a mean birth weight under 2,000 g. Infants who develop IRDS frequently have lower alpha1AT levels than those who do not develop the syndrome, independently from body weight. On the basis of serum alpha1AT quantitation, newborn infants may be separated into two groups, characterized respectively by concentrations above or below 150 mg%. From our data, it appears that if the group with an alpha1AT concentration lower than 150 mg% is phenotyped, it is possible to differentiate infants with a high risk of fatal IRDS from individuals with a "pathological" phenotype.


Endovesical transurethral electrostimulation in the rehabilitation of neurogenic bladder in children. 4 Years clinical experience. 4 years of experience in the rehabilitation of neurogenic bladders by endovesical electrostimulation (ES) with 31 children permits us to judge the value and limitations of this treatment. 8 children (26%) reacted favorably to treatment and acquired continence approaching normal. 12 children (38%) showed modifications of bladder-sphincter equilibrium, without acquiring socially sufficient continence. 10 children (33%) showed no response to treatment. In 1 child ES may have precipitated bladder decompensation. An analysis of clinical, manometric, and radiological results in comparison to the underlying pathology permitted us to define the contraindications to ES therapy, criteria which should in the future permit a more efficient selection of patients.


Prosthetic valve hemolysis aggravated by gastrointestinal bleeding. A case is presented of a patient with 2 prosthetic valves and a mild chronic hemolytic anemia, in whom the hemolysis was suddenly aggravated by the occurrence of an acute gastrointestinal hemorrhage. This increased hemolysis was readily reversed by transfusion. It is postulated that the pathogenetic mechanism of this process was the increased direct crushing trauma and turbulence, both of which resulted from the tachycardia initiated by the gastrointestinal hemorrhage. The importance of avoiding or correcting tachycardia in patients with prosthetic valves is emphasized.


The production of hemopoietic growth factors by PHA-stimulated leukocytes. Conditioned medium from cultures of phytohemagglutinin-stimulated leukocytes has been assessed for its content of factors able to stimulate either colony formation by human granulopoietic progenitor cells, or 3H-thymidine incorporation by peripheral leukocytes from leukemic patients. The cell concentration, concentration of phytohemagglutinin and time required for optimal production of factors were investigated. Production of both classes of factors was found to be insensitive to inhibitors of cell proliferation and DNA synthesis, but sensitive to inhibition of protein synthesis by cycloheximide. The production of colony stimulating activity showed a markedly greater sensitivity to cycloheximide than did production of thymidine-incorporating activity. For cycloheximide and most of the other inhibitors studied, factor production by leukemic cell populations was affected to a somewhat greater extent than production by normal leukocytes. Separation by velocity sedimentation of the cells responsible for production provided no evidence for distinct classes of cells producing these two classes of factors.


In vitro characteristics of childhood leukemic monoblasts. Leukemic monoblasts obtained from three children were evaluated for their in vitro physiologic characteristics. These monoblasts were alpha naphthyl acetate esterase positive, exhibited glass adherence, had IgG membrane receptors, phagocytized latex particles with subsequent nitroblue tetrazolium (NBT) reduction, and matured to macrophages in vitro, either when adherent to glass slides or in feeder layer suspensions. Employing the double layer in vitro bone marrow culture technique, leukemic marrows from these children failed to produce colonies in culture in the presence or absence of normal feeder layers. Varying the concentration of leukemic cells in the feeder layers failed to augment colony growth from normal bone marrows. Although monoblasts were observed to mature to macrophages in vitro, no increase in colony stimulating factor activity was observed with aging of the feeder layers. These studies suggest that childhood leukemic monoblasts lack the ability to produce colony stimulating activity, differing in this respect from monoblastic leukemia in adults.


A longitudinal study of T and B lymphocytes from a three-year-old patient with severe combined immunodeficiency (SCID) in 'gnotobiotic protection'. Fluctuations in the percentages and absolute numbers of T and B lymphocytes were observed in the peripheral blood of a patient with severe combined immunodeficiency maintained in a gnotobiotic environment. Up to 24 months of age, 72-86% of the lymphocytes had surface membrane immunoglobulin (SMIg), 37-47% bore a receptor for C3(EAC-RFC), and 3-12.5% formed spontaneous rosettes with sheep erythrocytes (E-RFC). These values persisted until 30 months, after which shifts in the percentages and absolute numbers of T and B cells were observed. A significant decrease in the proportion of SMIg-bearing cells to 20-40% (169-405 mm3), and EAC-RFC to 10.5-39% (114-259 mm3), was accompanied by a general increase in the proportion of T cells to 19-60% (141-1026 mm3), representing a lymphoid subpopulation approach to normal levels.


Hematopoiesis on cellulose ester membranes (CEM). I. Functional characteristics of cells comprising the hematopoietic microenvironment. Cellulose ester membranes (CEM) implanted into the peritoneal cavity of mice rapidly became coated with cells of peritoneal origin. Up to 56% of the cells, at a peak point 3-5 days after implantation, showed cell membrane receptors for complement and cytophilic immunoglobulin. A similar proportion of cells from CEM phagocytized yeast particles in vitro. When studied in situ, rosettes with C3b and IgG coated erythrocytes were formed by 23% of the cells coating CEM. A decreasing percentage of cells with monocyte-macrophage characteristics were detected between 5 and 17 days. CEM removed at 2 or 6 weeks after peritoneal implantation enriched tissue cultured media with colony stimulating factor which supported the growth of granulopoietic colonies in softagar culture. Mice given 59iron and 99technetium sulfur colloid i.v. showed substantial uptake of both isotopes by the CEM but the 59iron uptake could not be suppressed by hypertransfusion. Surface hematopoietic colony formation on CEM was studied 1-14 days after implantation. A peak colony number occurred at 5 days and the fell off slightly by 2 weeks. These studies indicate that the hematopoietic microenvironment of peritoneally implanted CEM contains a major sub-population of cells with monocytemacrophage features. The hematopoietic microenvironment was well-maintained even though the percentage of monocyte-macrophage marked cells decreased indicating that the microenvironment is not solely dependent upon monocyte-macrophages.


[Effect of triiodothyronine and methylthiouracil on the enzymatic activity of biogenic amine metabolism in rat brain synaptosomes]. Triiodothyronine injections are shown to lower DOPA decarboxylation and 5-hydroxytryptophan decarboxylation activity and also to decrease the intensity of serotonin and norepinephrine oxidative deamination in the rats' brain synaptosomes. Hypothyroidism produced by 6-methylthiouracil administration was accompanied by intensification of the DOPA-decarboxylase activity and monamine oxidase transformation into serotonin and norepinephrine with no parallel occurring essential changes in the 5-hydroxytryptophan decarboxylation.


[Activity of antifibrinolytics when administered using a direct current]. Tests conducted on rabbits demonstrated a high effectiveness of fibrinolysis inhibitos (tracilol, E-aminocapronic acid) with their electrophoresis as compared with their intravenous administration. The absence of any influence of direct current on the activity of contrical, tracilol and E-ACA was ascertained.


Glucocorticoid regulation of rat thymus RNA polymerase activity: the role of RNA and protein synthesis. Treatment of rat thymus cells with the glucocorticoids cortisol and dexamethasone resulted in the stimulation of RNA polymerase B activity within 10 min of steroid addition. This early effect was followed by the inhibition of both RNA polymerase A and B activities. These effects were glucocorticoid-specific and were inhibited by the antiglucocorticoid cortexolone. The inhibitory effect of dexamethasone on RNA polymerase A activity was abolished by prior treatment of the cells with alpha-amanitin, cordycepin or cycloheximide, but cycloheximide was only capable of inhibiting the steroid effect measured at 3 h if added within 10--20 min after steroid addition. Cycloheximide had no effect on the steroid-mediated inhibition of RNA polymerase B activity. Control RNA polymerase A activities were unaffected by the presence of inhibitors of RNA and protein synthesis. It is concluded that the inhibition of ribosomal RNA synthesis by glucocorticoids is dependent on protein synthesis, but that basal RNA polymerase A activity in rat thymus cells is not stringently coupled to protein synthesis.


Spleen size and portal hypertension in cirrhosis. In 210 patients with cirrhosis of the liver there was no correlation between spleen size and the rise in portal pressure, whether measured directly or assessed clinically from the size of varices on barium swallow and the presence of ascites. Despite this, there was a correlation between the occurrence of bleeding as a presenting symptom and the size of varices assessed radiologically.


Distinct recognition of influenza virus hemagglutinin and H-2 antigens by cytotoxic thymus derived lymphocytes. Cytotoxic thymus derived (T) lymphocytes were readily detected in BALB/c and C3H mice during infection with influenza A (H0N1, H3N2, Heq1Neq1) and B viruses. T cell mediated lysis was specific for H-2 compatible target cells infected with the same strain of virus used to immunize the mice. The viral specificity was found to be related to the hemagglutinin antigen by the use of parent strains (H3N2 and HeqNeq) and their recombinant viruses which included the antigenic hybrids for hemagglutinin and neuraminidase (H3Neq1 and Heq1n2). In addition, cytotoxic T cell mediated lysis of influenza infected cells was demonstrated to be restricted to syngeneic cells and cells sharing the same H-2 haplotype. Thus, T cell mediated lysis of influenza infected cells appears to involve the distinct recognition of viral and H-2 antigens on the infected cell by the cytotoxic T cell.


Effects of cell mediated immunity in influenza virus infection in mice. The role of cell mediated immune responses in recovery of mice from influenza virus infection was studied by immunosuppression and by adoptive immunization. Virus infections persisted longer and produced more severe lung lesions in animals injected with anti-thymocyte serum (ATS), and immunosuppressed animals failed to mount a serum HI antibody response. Mice injected with ATS after the 4th day of infection produced antibody in titers equivalent to those of control animals but still did not recover from infection as rapidly. Passive immunization with antibody late in infection did not facilitate clearance of virus from ATS injected animals. Adoptive transfer of spleen cells obtained from syngeneic animals sensitized intraperitoneally resulted in more rapid clearance of virus and less severe lung lesions in infected recipient animals. This effect was associated with a lower antibody response in recipient mice. Adoptive immunization was still effective after depletion of beta lymphocytes from the transferred cell population. These effects correlate with in vitro assays of the cytotoxic T cell response and antibody forming cell response of sensitized mice. From these observations we conclude that T lymphocytes may play a role in recovery from influenza virus infection by mechanisms other than helper effects.


T cells, precocious aging, and familial neoplasia. A 15-year-old girl presented with precocious aging and was found to have low levels of active and total T cells. Family history revealed a high familial incidence of cancer on both the maternal and paternal sides, and activ T cell levels were found to be low in several living family members. The patient developed osteogenic sarcoma 13 months after initial study. Since our previous studies have reported low active and total T cells in patients with cancer, the present results suggest that subjects with low active T cells should be monitored frequently to detect possible neoplasia in it early stages. They also suggest that impaired cellular immunity in humans is associated with, if not the cause of, accelerated aging.


T-lymphocyte cytotoxicity to HBsAg-coated target cells in hepatitis b virus infection. The cytotoxic effect of peripheral lymphocytes on chicken red blood cells (ChRBC) coated with purified hepatitis B surface antigen (HBsAg) has been studied as an in vitro parameter of cell-mediated immunity in acute and chronic infection with hepatitis B virus. Using this technique, the mean cytotoxic index of lymphocytes from patients with acute hepatitis B (29.13 +/- 20.88) was significantly higher than that obtained with lymphocytes from control subjects (6.53 +/- 3.75). Only 33.3% of the patients with HBsAg-positive chronic active hepatitis exhibited lymphocyte cytotoxicity to HBsAg-coated target cells and the mean cytotoxic index (11.66 +/- 6.60) was in these cases significantly lower than that found in acute hepatitis B. Healthy chronic carriers of HBsAg failed to show lymphocyte cytotoxicity to target cells. The effector cells detected in acute hepatitis B by this in vitro assay have been demonstrated to be T-lymphocytes, as T-cell depleted subpopulations lacked cytotoxic activity. Target cell lysis could be blocked by addition of HBsAg-coated unlabelled ChRBC as well as of purified HBsAg in the culture tubes. It is suggested that damage to the liver cells in acute hepatitis B is related to the presence of cytotoxic T-lymphocytes reacting with HBsAg on the surface of infected hepatocytes. An inadequate lymphocyte response to the antigen may be responsible for the persistence of the infection in the liver with varied clinical manifestations and associated hepatic lesions.


K-lymphocytes (killer-cells) in Crohn's disease and acute virus B-hepatitis. Total lymphocyte counts, B-, T-, C'3 receptor-bearing lymphocytes, and K-cell activity were studied in peripheral blood in patients with Crohn's disease and inflammatory liver disease. Patients with active untreated Crohn's disease and acute virus B hepatitis exhibited a markedly increased K-cell activity measured in a plaque assay when compared with normal controls (P less than 0.01). Patients with immunosuppressive treated Crohn's disease, HBsAg-positive chronic active hepatitis, and cirrhosis of the liver showed only a slight increase of K-cell activity (P less than 0.01). In the postacute phase of hepatitis (four to 12 weeks from onset) K-cell activity fell to normal levels. The number of B-lymphocytes showed a relative and absolute decrease in all groups of patients. With the exception of patients with acute HBsAg-positive hepatitis and the post-acute phase of hepatitis all the other groups showed statistically decreased absolute numbers for C'3 receptor-bearing lymphocytes. The significant decrease in K-cell activity and the number of T-lymphocytes in Crohn's disease treated with immunosuppressive drugs was interpreted as an effect of azathioprine and prednisone on these lymphocyte subpopulations.


Reduction in peripheral blood K cells and activated T cells in primary biliary cirrhosis. Lymphocyte subpopulations were measured in the peripheral blood of 13 patients with primary biliary cirrhosis (PBC), 13 with chronic active hepatitis (CAH), and 16 age- and sex-matched normal subjects. A significant, relative and absolute, reduction in activated T cells and K cells, compared with that in normal subjects, was found in PBC but not in CAH. The reduction in K cells observed in PBC was found to be accompanied by a parallel decrease in lymphocyte-mediated antibody-dependent cytotoxicity.


Sensibility of the rectum to distension and the anorectal distension reflex in ulcerative colitis. The maximum volume of air tolerated within a rectal balloon was less in colitic patients than in normal subjects (P less than 0.001). Smaller volumes were tolerated by patients with a spontaneously bleeding mucosa than by those with less severe inflammation (P less than 0.001). Severe urgency of defaecation with incontinence was experienced by about half those with spontaneous mucosal haemorrhage but was infrequent among other colitics. Those patients with inactive colitis and a reduced maximum tolerable volume tended to have a smaller rectal size than those who could tolerate a normal volume within the rectum. Reflex relaxation of the internal anal sphincter on distension of the rectum appeared normal in patients with colitis.


[Endometrial cytology and copper containing intrauterine devices]. Copper-containing intrauterine devices (IUDs) for contraception are increasingly used within recent years. IUDs in situ provoke a non-specific inflammatory cell reaction, particularly at the glandular epithelium of the endometrium. The cellular changes include swelling of the cytoplasm and of the nuclei, the latter showing chromatin clumping, prominent nuclei and a pseudoeosinophilic reaction. Cytoplasmic changes exhibit vacuolation, incorporation of leukocytes and plasmolysis varying in degree. The local inflammation also accounts for the side effects associated with the device, such as uterine bleeding and endometritis. Cytomorphologically, the differentiation between cellular inflammation through IUDs and inflammatory metaplasia is difficult, particularly if the investigator has no information about the presence of an IUD. Intrauterine application of proteinase inhibitors can be helpful in differential diagnosis of such IUD-mediated cellular changes.


Effect of phospholipases on factor-VIII activity. The effect of the enzymes phospholipases C and D on Factor VIII were investigated. Phospholipase D was found to activate the partially purified intact Factor-VIII molecule maximally at a final concentration of 0.6 U/ml. Neither the dissociated small molecular weight component nor the high molecular weight component were activated. Phospholipase C, on the other hand, inactivated both the intact and the dissociated Factor-VIII molecule. Phospholipase D, however, had no effect on the haemophilic cryoprecipitate or the partially purified haemophilic Factor VIII. The implications of these results for the genetic control of the Factor-VIII molecule are discussed. In this connection, haemophilia A is hypothesized to be caused by an X-linked enzyme effect that impairs phospholipid assembly of the Factor-VIII protein, whereas von Willebrand's disease might be due to a structural defect of the Factor-VIII protein.


Induction of hamster T cell-specific antiserum in rabbits using hamster brain tissue. Antiserum specific for hamster thymus-derived lymphocytes, prepared by immunizing rabbits with hamster brain tissue, was cytotoxic to hamster thymocytes greater than lymph node cells greater than spleen cells, while virtually non-reactive against bone marrow. This antiserum inhibited spleen cell response to the T cell mitogen, phytohemagglutinin, but not to lipopolysaccharide, a B cell mitogen.


Alterations in the mitogen responsiveness of lymphocytes from mice bearing Moloney sarcoma virus induced tumors. The effects of Moloney Sarcoma Virus (MSV) induced tumor growth dynamics on the blastogenic responsiveness of lymphocytes from BALB/c mice were investigated. Lymphocytes from spleen, thymus and lymph node pools were tested for responsiveness to phytohemagglutinin (PHA) and concanavalin A (Con A). Results showed a significant decrease in PHA-induced blastogenesis of all lymphocytes tested at the time of maximal tumor volume, with a return to normal responsiveness as the tumor regressed. In contrast, a differential dose dependent Con A response occurred in spleen and thymus lymphocytes. A decreased 3H-thymidine uptake occurred at optimal Con A dose, correlating with the PHA decrease. However, at a lower Con A dose an increased response was observed, beginning shortly before the PHA depression and continuing until regression of tumor began. This phenomena was not observed in lymph node lymphocytes. Based upon these observations, we suggest that the cell or cells responsible for the transient suppression of PHA responsiveness may be Con A responsive T lymphocytes.


Embryonic mouse thymus development: stem cell entry and differentiation. The entry and subsequent division and differentiation of blood-borne stem cells within the embryonic thymus has been studied in a system where the inflowing cells were either chromosomally or antigenically marked. H-2 positive stem cells enter the thymus within 5 days of grafting but may wait for up to 14 days before proliferating to give rise to Thy-1 positive thymocytes.


The effect of protease inhibitors on leucocyte migration inhibition to tuberculin extract (P.P.D.). The protease inhibitors, Epsilon Amino Caproic Acid (E.A.C.A.) and Aprotinin were found to significantly increase the inhibition produced by P.P.D. in the Leucocyte Migration Test (L.M.T.). Dose-response curves showed that the most effective concentrations were for E.A.C.A. 0.01 M and Aprotinin 100 I.U./ml. In a series of 29 subjects with a range of Heaf reactivity this finding was consistent providing that there was a significant inhibition to P.P.D. alone. At these concentrations, cell viability was unaffected and the effect of the drugs could be reversed by washing. The exact mode of action of these inhibitors in the L.M.T. requires further elucidation.


Immunological tolerance to lymphocytic choriomeningitis virus in neonatally infected virus carrier mice: evidence supporting a clonal inactivation mechanism. Previous studies have shown that no cell-mediated immunity against LCM virus-infected cells can be detected in neonatally established LCM virus carrier mice suggesting that they are immunologically tolerant to virally-altered cell membrane antigens. In this communication experiments are described aimed at analyzing the mechanism. Virus-specific cell-mediated immunity was assessed by 51Cr release and target cell reduction assays. Attempts to demonstrate cells in spleens of CBA/J carrier mice able to suppress in syngeneic recipients the induction or the effector phase of the cytotoxic T-cell response against LCM virus-infected cells were unsuccessful. Also, no factors were detected in CBA/J and C57BL/6J carrier mice, either spleen cell-associated or free in the circulation, which would block the activity of cytotoxic T-lymphocytes against LCM virus-infected syngeneic target cells. The results indicate that inability of LCM virus carrier mice to act immunologically against virus-infected target cells is due to deletion or irreversible inactivation of T lymphocytes carrying receptors for virally altered cell membrane antigens.


Immune effector cell activity in canines: failure to demonstrate genetic restriction in direct antiviral cytotoxicity. Experiments were undertaken to establish whether the cytotoxic activity of canine immune effector cells against viral antigens was affected by the genotype of the target cell (genetic restriction). Puppies from three different breeds were infected with vaccinia virus, and the peripheral blood leukocytes were collected at various times for measurement of their cytotoxicity against autologous and heterologous vaccinia virus-infected and uninfected skin fibroblasts. In all cases cytotoxicity only occurred against virus-infected targets, and there was no consistent evidence of preferential killing of autologous targets. Several indirect approaches were used to demonstrate that direct, presumably T cell, cytotoxicity was being measured rather than antibody-dependent cell cytotoxicity. On the basis of the evidence from cross mixed-lymphocyte assays and cell-mediated lympholysis assays, the dogs were shown not be be identical with respect to their histocompatibility antigens. The significance of our findings to the phenomenon of genetic restriction as observed for mouse-derived immune effector cells is briefly discussed.


Characterization of the immunoprotective antigen of ribosomal preparations from Haemophilus influenzae. This investigation was designed to characterize the immunoprotective antigen of ribosomal preparations from Haemophilus influenzae. The ribosomes that elicited 80 to 90% protection contained 25% protein and 75% ribonucleic acid but did not contain any detectable hexoses. The immunodiffusion and hemagglutination inhibition tests also failed to demonstrate that the capsular material (polyribose phosphate) was in ribosomal preparations. Treatment of ribosomes with ribonuclease degraded 78% ribonucleic acid but did not affect the immunogenicity of such preparations. The proteolytic enzymes reduced the immunogenicity of ribosomes corresponding to the amount of protein degraded. The protection elicited by ribosomal protein extracted with 2-chloroethanol was comparable to that induced by intact ribosomes. In contrast, the low levels of protection observed by immunization with phenol-extracted ribonucleic acid were dependent on the amounts of contaminating protein. Finally, immunogenicity of ribosomal ribonucleic acid and protein was abrogated by treatment with proteolytic enzymes. These results clearly indicate that the protein associated with Haemophilus ribosomes is the major immunoprotective antigen.


Characterization of the human natural killer (NK) cell in blood and lymphoid organs. The human natural killer (NK) cell in peripheral blood is a non-thymus-dependent, (Fc+ C3-), Ig-bearing lymphocyte and is probably identical to the human K cell, although not requiring the Fc receptor, in contrast to the K cell, for the lytic process. The human NK cell in tonsil and lymph node, on the other hand, is a non-Ig- bearing T lymphocyte, lacking both Fc and C3 receptors.


Development and persistence of cytolytic T lymphocytes in regressing or progressing Moloney sarcomas. Intratumoral T lymphocytes were recovered sequentially after induction of regressing or progressing Moloney sarcomas in BALB/c mice and were assayed quantitatively for their ability to kill specifically the tumor (MSC) cells used for induction. The cytolytic activities of the two lymphocyte populations described two distinct biphasic kinetic profiles that were similar in amplitude and duration but separated from each other by 4-6 days. In progressing neoplasm, there was a rapidly occuring accumulation of T lymphocytes highly cytolytic for MSC cells. This response, however, was not sustained and disappeared in association with the onset of unchecked tumor growth. In contrast, T lymphocyte cytolytic activity developed more slowly in regressing sarcomas and attained peak levels coincident with the beginning of tumor regression. Similar changes in cytolytic activity characterized T lymphocytes in lymph nodes draining tumors. When cultured in vitro for 4 days, non-cytotoxic T lymphocytes from regional lymph nodes draining progressing sarcomas regained very high levels of cytolytic activity. Such restitution was diminished, however, if MSC cell lysates, macrophages or macrophages fed MSC cell lysates were present during the culture period. These experiments provided presumptive evidence that T lymphocyte-mediated cytolytic activity was lost in progressively growing Moloney sarcomas as a consequence of suppression in vivo of the genesis and/or functional expression of cytolytic T lymphocytes, perhaps by macrophages and/or soluble tumor antigen.


Negative pion irradiation of mammalian cells. II. A comparative analysis of cell-cycle progression after exposure to pi-mesons and cobalt gamma-rays. The effect of negative pi-mesons on cell-cycle progression of exponentially-growing V 79 monolayer cultures was examined by flow-microfluorometric analysis. Comparison with proliferative changes of the same cell-line after cobalt gamma-irradiation yielded r.b.e. values (relative biological effectiveness) of 2.2 for both, G2+M accumulation and division delay after irradiation at the pion peak. A dose-dependent prolongation of S-phase was found after Co gamma-irradiation which, however, was not detectable after pi-exposure. This observation is so far the only qualitative difference demonstrated in the effects of the two types of radiation.


Implication of protein aromatic amino acids in concanavalin A-carbohydrate interactions. A radiolytic study. Pulse radiolysis and 60Co gamma-radiolysis have been used to study the effects of ionizing radiation on the glycogen-precipitating activity of the protein concanavalin A. Both eaq- and OH. react extremely rapidly with the protein and contribute to the observed loss of concanavalin A activity. Evidence is also presented that (Br)2-. and (SCN)2-. react with with tyrosine and tryptophan residues in concanavalin A, which is also accompanied by loss of glycogen precipitation activity.


Radionecrosis of normal tissue: studies on mouse tails. Studies on the radionecrosis of mouse tails demonstrate the following modifications to the dose necessary for necrosis in 50 per cent of tails (the ND50): (a) There is very little reduction in ND50 values for irradiated lengths of tail from 2 cm to almost the whole tail, but there is a sharp increase in dose for lengths less than 1.5 cm. (b) The ND50 is high for unanesthetized mice irradiated in air, due to tissue hypoxia. (c) The hypoxia can be reduced by varying amounts by applying heat to the tail, or by flowing oxygen over the tail surface, or by anesthetizing the animal. (d) The ingress of oxygen through the surface can be reduced by placing a clamp round the proximal tail. These features are discussed with reference to the state and possible position of the target cells, and to the use of this assay technique in comparative studies.


Lethality in mammalian cells due to hyperthermia under oxic and hypoxic conditions. From several studies of hyperthermia there have been reports that hypoxic cells are more sensitive to heat than their oxic counterparts. Experimental techniques in this investigation eliminate the effect of pH, trypsinization and cell attachment, when assaying the effects of hyperthermia on cells. Under hypoxic conditions, HeLa S3 and Chinese hamster cell-lines do not have an increased sensitivity to heat compared with oxic cells. HeLa S3 cells are protected against heat by hypoxia. Light-microscopy indicates the rupture of the plasma membrane, occasional nuclear budding, membrane vesicles and granulation of cell contents after heating at 43 degrees C for 3 hours. Scanning electron micrographs show that cells are more rounded after heat treatment and that there is an accompanying decrease in the number of microvilli, suggesting that the mechanism of cell attachment is affected. Heated cells should be delicately handled and subjected to the minimal trauma so that an accurate comparison of survival can be made.


Cytochemistry of blood cells in two West African amphibians. Lipid, glycogen, peroxidase, acid and alkaline phosphatases were studied in the cells of blood smears in the toad and frog. While lipid and glycogen were largely confined to granulocytes and monocytes, peroxidase activity was intense in all cell types except lymphocytes in both species.


Membrane-bound, pyridine nucleotide-independent L-lactate dehydrogenase of Rhodopseudomonas sphaeroides. Rhodopseudomonas sphaeroides has a pyridine nucleotide-independent L-lactate dehydrogenase associated with the membrane fraction of cells grown either aerobically or phototrophically. The dehydrogenase is present in cells grown on a variety of carbon sources, but at levels less than 20% of that found in cells grown with DL-lactate. The dehydrogenase has been purified 45-fold from membranes of strain L-57, a non-photosynthetic mutant, by steps involving solubilization with lauryl dimethylamine oxide and three anion-exchange chromatography steps. The purified enzyme was specific for the L-isomer of lactate. The Km of the purified enzyme for L-lactate is 1.4 mM, whereas that of the membrane-associated enzyme is 0.5 mM. The enzyme activity was inhibited competitively by D-lactate and non-competitively by oxalate and oxamate. Quinacrine, a flavin analog, also inhibited the activity. The inducible enzyme may serve as a marker of membrane protein in studies of membrane development.


alpha-Amylase from human pancreatic juice as an electrophoretically pure isozyme. Human pancreatic juice collected antiseptically and strictly free from other body fluids and juices, was examined for the isozyme composition of alpha-amylase. Unlike those of the extract of and exudate from pancreatic tissue, the composition of alpha-amylase isozyme in the secreted juice was simple and consisted of only two isozymes, a major one and a minor one, and the reason for difference in the isozyme composition is discussed. The major alpha-amylase isozyme was obtained in a crystalline state with a molecular weight of 5.4-5.5 X 10(4).


Bacteremia associated with Haemophilus influenzae type e biotype 4. Haemophilus influenzae type e biotype 4 was isolated from a single antemortem blood culture obtained from a 60-year-old, white male with abdominal carcinomatosis.


Detection of capsulated Haemophilus influenzae in chest infections by counter current immunoelectrophoresis. The application of counter current immunoelectrophoresis to the detection of Haemophilus influenzae capsular antigen in sputum is described. The method, technically simple, provided results within 30 minutes. H. influenzae capsular antigen was detected in 12% of patients and in 54.8% of the H. influenzae strains isolated. The test was not impaired by prior antibiotic therapy.


Studies on the mechanism of action of guinea pig lymphotoxin. I. Membrane active substances prevent target cell lysis by lymphotoxin. The effect of various pharamacologic agents on lysis of L cells by guinea pig lymphotoxin (LT) was studied. Among the many drugs tested, only those that affect plasma membrane functions were found to interfere with the cytolytic action of LT. Dimethyl sulfoxide or lidocaine potentiated L cell resistance against lysis. Stronger protection was provided by ouabain. Addition of ouabain to cells previously injured by LT was also effective in reducing cell death. Attempts to detect metabolic disturbances in cells before LT cytolysis were unsuccessful. The biosynthetic rate of DNA, RNA, or protein, and the total cellular content of ATP were not significantly changed in LT-treated cells. The results suggest that LT disturbs some plasma membrane functions of the target cell, perhaps ion transport systems, and consequently induces ionic imbalances between the intra-and extracellular milieu which eventually cause cell death.


Studies on the mechanism of action of guinea pig lymphotoxin. II. Increase of calcium uptake rate in LT-damaged target cells. In the previous paper it was suggested that the primary action of guinea pig lymphotoxin (LT) involved creation of ionic imbalances within the target L cells. The nature of these ionic disturbances is explored in this paper. The exogenous addition of CaCl2, but not KCl or NaCl, inhibited the cytotoxic action of LT. Cellular uptake rates of 45Ca++, but not 86Rb+, increased in LT-damaged L cells. The factor responsible for increasing the 45Ca++ uptake rate cochromatographed on a hydroxyapatite column with the cytotoxic activity of LT. Ouabain prevented the LT-mediated lysis and, concomitantly, depressed the LT-induced increase of 45Ca++ uptake rate. The LT-damaged L cells excluded trypan blue to the same extent as the normal cells. The addition of LT to and LT-resistant L cell mutant affected neither the 45Ca++ uptake rate nor the viability. From these observations, damage to the calcium transport system in the L cell plasma membrane is proposed as a mechanism of LT action.


Low affinity E-rosette formation by the human K cell. Human T lymphocytes were separated into two subsets on the basis of relative affinity for sheep red blood cells (E), and these T cell fractions were examined for cytotoxic reactivity against antibody-sensitized Change liver cells (ADCC). High affinity E-rosette-forming cells (E-RFC) (55 +/- 6% of peripheral blood mononuclear cells), capable of rosette formation despite elevated temperatures of incubation (29 degrees C) and a limited concentration of E, contained few antibody-dependent cytotoxic cells (K cells). In contrast, low affinity E-RFC (23+/-7% of mononuclear cell suspensions) requiring cool temperatures of incubation (4 degrees C) and an excess of E to form rosettes, were highly enriched for ADCC activity. The majority of K cells exhibited low affinity interactions with E. T cells in thymus, tonsil, and lymph node formed high affinity E-rosettes and exhibited little reactivity in ADCC. Only peripheral blood and spleen contained easily identified low affinity E-RFC and anti-body-dependent cytotoxic cells. The proportion of low affinity E-RFC in the peripheral blood of normal subjects correlated closely with reactivity in ADCC, making it possible to predict cytotoxic potential for the E-rosette pattern. These data indicate that the human K cell may belong to a previously unappreciated but functionally important subset of thymic dependent mononuclear cells.


Mucus viscoelasticity and mucociliary transport rate. The aim of this research was to define in quantitative terms the relationship between the viscoelastic properties of mucus and the velocity at which it can be transported by a ciliated mucosa. To this effect, tests were performed on frog palates with normal beating cilia which were washed free of endogenous mucus. Experimental sputum samples encompassing a wide range of viscoelastic properties were applied to the palates, and their transport velocity was measured under a dissecting microscope. The results indicated that the fastest transport, per palate, was achieved by sputa whose Newtonian viscosity and linear elastic modulus fell within the range of 1,000 to 3,000 poises and 10 to 25 dynes/cm2, respectively. The transport velocity decreased when the sputum viscoelasticity was above or below this optimal range. The latter condition is of particular importance since it indicates that marked fluidification may hinder rather than promote mucus clearance.


Characterization of p-aminobenzoic acid transport across the rat intestine. The intestinal absorption of PABA, a member of the vitamin B complex, was characterized in vivo and in vitro in the rat by the segmental intestinal perfusion and everted gut sac technique. Net PABA absorption was directly proportional to substrate concentration, and saturation of absorption did not occur with increasing concentrations of PABA (1 to 50 mM), indicating a nonsaturable process. Jejunal and ileal absorption rates were similar and were not influenced by the presence of glucose or the absence of sodium in the test solution. Similarly, 14C-PABA transport in vitro was nonsaturable and proportional to PABA concentration (0.05, 0.5, 1, 10, and 50 mM). It was not inhibited by ouabain or other PABA analogues such as folic acid and benzoic acid. These studies indicate that PABA, a vitamin B cofactor, is absorbed by a nonsaturable, sodium-independent process, which characterizes passive diffusion and is similar to the absorption of other vitamin B members.


Immunological T-cell memory in the in vitro-induced experimental autoimmune orchitis: specificity of the reaction and tissue distribution of the autoantigens. Immunological memory has been induced in vitro against testicular autoantigens by priming normal rat T lymphocytes against autologous testis cells, and by permitting the isolated blast cells to revert back to small secondary lymphocytes (secondary EAO cells) in the absence of the priming antigen. The secondary EAO cells vigorously respond in a secondary response when reconfronted with syngeneic testis or lymphoid cells. Their responsiveness to nonself stimulator cells is, however, reduced. Secondary cells derived from concanavalin A-stimulated blasts, do not show that pattern of specificity. The specificity of the secondary EAO cells is definite, and cannot be affected by further culture on allogeneic fibroblasts, which are antigenic for unprimed T lymphocytes. At least part of the autoantigens are determined by the major histocompatibility gene complex (MHC). Factors provided by the culture system do not appear to determine the specificity of this reaction. Only minor cell populations can restimulate secondary EAO cells. One of these populations is presumably phage-like cells within the lymphoid populations can elicit a secondary EAO response. Thus, the autoantigens relevant in the secondary EAO response are either MHC antigens restricted to these testicular and lymphoid subpopulations, or MHC antigens recognized in conjunction with organ-specific non-MHC determinants.


Changes in suppressor mechanisms during postnatal development in mice. The activity of suppressor cells from spleens of mice of varying ages was assessed by their addition to cultures of normal or SRBC immune spleen cells together with a challenge of SRBC. 1-wk and adult spleen cells were highly suppressive of the secondary in vitro antibody response to SRBC. 3-wk spleen cells were less active in suppressing this response. The nature of the suppression and the character of the suppressor cells changed in this period. Whereas adult spleen cells demonstrated specificity, 1-wk cells nonspecifically suppressed all responses tested. Further, unlike adult suppressor cells (which are Thy.1.2 positive), 1-wk suppressor cells are insensitive to anti-Thy.1.2 treatment in this system. Both cells are nonadherent to glass beads and nylon wool and are undetectable in the normal thymus.


Human erythroid burst-forming unit: T-cell requirement for proliferation in vitro. Human mononuclear leukocytes were fractionated into populations of null, T and B cells by immunoabsorbent column chromatography followed by E-rosette formation and purification of T cells by differential centrifugation and osmotic lysis. The unfractionated and fractionated cell populations were first separately cultured for 14 days in plasma clots in the presence of two international units erythropoietin. Typical erythroid burst-forming unit (BFU-E)-derived colonies grew in the unfractionated cell cultures but not from T- or B-cell cultures. BFU-E colonies grew in null cell cultures but most of the colonies were small and variably hemoglobinized with less than three subcolonies. When intact T cells were added to null cells and cocultured, many typical large BFU-E colonies with more than 10 well homogenized subcolonies appeared. Increasing numbers of large BFU-E colonies in null cell cultures were induced by stepwise addition of T cells but not by the addition of B cells. A conditioned medium in which T cells had been induced to divide by tetanus toxoid substituted for intact T cells in this T-cell-dependent BFU-E colony formation observed in null cells. These findings demonstrate that the BFU-E, a committeded erythroid stem cell, resides in the null cell fraction of peripheral blood, but its proliferative capacity and differentiation in vitro requires a soluble product of T cells. Such experiments now permit a new approach to the assessment of various disorders of erythropoiesis. Erythroid hypoplasia in a particular case may be due to dysfunction of the committed precursor cell or to a failure of a helper effect induced by T cells.


Presence of anti-Sm reactivity in autoimmune mouse strains. The investigation of the fine specificities of antinuclear antibodies (ANAs) has been fruitful in terms of the nosology and immunopathogenesis of human autoimmune syndromes. Particular reactivities serve as "markers," in that patients with certain syndromes have a much higher incidence of such ANAs than do patients with other diseases. In this category is the almost exclusive against the nuclear acidic protein Sm. Reactivity to Sm can be detected by precipitation in agar, complement fixation, or passive hemagglutination (1,2). Autoimmune mouse strains have also provided a fertile field for the investigation of the basic phenomena of self-activity. In particular, the NZB strain and its hybrid NZB x NZW have been considered excellent models for human SLE and have therefore been studied in great detail (3,4). In addition, Murphy et al at The Jackson Laboratory, Bar Harbor, Maine, have developed several new inbred mouse strains that spontaneously develop SLE-like syndromes (5,6). These are the BXSB strain, which has a male dominant disease characterized by little antiative DNA antibody; the MRL/1, which develops massive, nonmalignant lymphadenopathy, associated with enormous increases in serum immunoglobulin levels and fulminant renal disease; and the MRL/n, which does not develop SLE-like disease until well into the 2nd yr of life, but like the MRL/1 develops high titers of ANA and fatal glomerulonephritis. The MRL/1 differs from MRL/n in only about 10 percent of its genome, including the gene responsible for the MRL/1's lymphoproliferation. In the current study, we have used the technique of double immunodiffusion (ID) in agarose with standard human reference sera (of known ANA specificity) to survey a large number of mice from the NZB x NZW, MRL/1, MRL/n, BXSB, and other strains. We report here the finding of the anti-Sm marker" antibody almost uniquely in MRL/1 and MRL/n animals. These two related strains may serve as experimental models to explore the mechanism stimulating the production of this unique autoantibody in SLE.


Induction of specific transplantation immune reactions using anti-idiotypic antibodies. B and T lymphocytes with reactivity against major histocompatibility antigens are known to express this immune potential via a display on the outer surface of antigen-specific, idiotypic receptors. Here, we show that anti-idiotypic antibodies directed against such receptors may serve as specific triggering agents of the idiotype-positive lymphocytes in the physical absence of foreign histocompatibility antigens. This was shown in vitro using normal or immune spleen T cells where anti-idiotypic antibodies would lead to the selective proliferation and development of antigen-specific cytolytic T cells as determined by short-time 51Cr release assays. Furthermore, purified anti-idiotypic antibodies in adjuvant administered in vivo to normal syngeneic animals could be shown to lead to production of high titers of specific alloantibodies. The present experiments were in most cases carried out using auto-anti-idiotypic antibodies as triggering agents. The present results thus lend further support to the concept that idiotype-anti-idiotype reactions may be normal parts of conventional immune processes with either stimulatory or inhibitory consequences, depending upon the prevailing conditions.


Good outcome in a catatonic patient with enlarged ventricles. In the differential diagnosis of the catatonic syndrome, the demonstration of an intracranial anomaly is often taken as evidence of irreversibility. We present the case of a 27-year-old white female with catatonia who was found to have enlarged ventricles on automatic computerized tomographic axial scan. She had a complete resolution of the catatonia and psychotic symptoms without any change in the size of the ventricles. This resolution occurred when the patient was treated for her ulcerative colitis with colectomy and the steroids she received for the colitis were gradually withdrawn. The discovery of a structural anomaly of the brain per se should not discourage the clinician from identifying and treating all other factors that might contribute to the catatonic syndrome.


5-hydroxytryptophan-induced myoclonus in guinea pigs. A physiological and pharmacological investigations. 5-Hydroxytryptophan (5-HTP) induces a characteristic behavioural syndrome of altered motor activity with muscle jerking in guinea pigs. Myoclonic jerking occurs synchronously in forelimbs and hindlimbs and is associated with a stereotyped electromyographic (EMG) pattern of a burst of activity lasting 40-50 msec in active muscles, followed by silence lasting 50-70 msec, followed by a further variable period of muscle activity. Such myoclonus may be induced also by the administration of L-tryptophan plus a monamine oxidase inhibitor (MAOI), or by agents acting as serotonin (5-HT) receptor agonists. The 5-HTP-induced syndrome is antagonised by a central decarboxylase inhibitor (NSD-1035) and by agents which block 5-HT receptors (methysergide and cyproheptadine). 5-HTP-induced jerking is abolished below the level of a spinal cord transection, but persists in decerebrate animals. No electroencephalographic (EEG) changes are seen preceding the muscle jerks. The clinical significance of this animal model of myoclonus is discussed.


Primary malignant schwannoma of the Gasserian ganglion. Report of two cases. The diagnosis, radiographic features, clinical presentation, and treatment of two cases of primary malignant schwannoma of the Gasserian ganglion are discussed. Radiographic differentiation from trigeminal neurinoma is not possible; however, erosion of the foramina favors a diagnosis of malignancy. Radiation therapy was successful in the management of both cases and is recommended for extensive lesions.


Endoscopy of Meckel's cave, cisterna magna, and cerebellopontine angle. Technical note. A new 1.45-mm endoscope is described that can be inserted through a thin-walled No. 16 needle. The instrument was used in 10 cadavers for endoscopic exploration of the cisterna magna, the C1-2 space, Meckel's cave, and the cerebellopontine angle. Its potential clinical application is discussed.


Inactivation of sodium channels: second order kinetics in myelinated nerve. 1. Kinetics of inactivation of sodium channels in myelinated nerve from Rana pipiens were studied at 4.5 degrees C using the voltage clamp technique of Dodge & Frankenhaeuser (1958).2. Potassium currents were blocked by cutting the internodes in 20 mM-TEA-Cl + 100 mM-KCl and by adding 12 mM-TEA-Cl to the external Ringer. Leakage and capacitative currents were subtracted electronically.3. Kinetics of recovery from inactivation of the sodium channels were studied by inactivating the channels with a large depolarizing prepulse and allowing the channels to recover at different potentials; the extent of recovery was measured by applying a test pulse at various times after the prepulse.4. Kinetics of development of inactivation were studied by two different methods. The first was to measure the decay of sodium current under a maintained depolarization. The second method was to measure the decay of the peak sodium current in a test pulse as a function of time after the onset of a maintained depolarization. These two methods yielded similar results for the kinetics of inactivation development.5. Contrary to expectations of the Hodgkin-Huxley formalism, the time course of recovery from and development of inactivation is not strictly exponential. Rather, recovery from complete inactivation shows an initial delay which depends on recovery potentials. Development of inactivation at a fixed potential exhibits at least two exponentials.6. The steady-state inactivation curve h(infinity)(E) is asymmetrical and is fitted better by 1/[1+exp (A(1)E+B(1)) +exp (A(2)E+B(2))] than by 1/[1+exp (AE+B)].7. Most of the above kinetic observation on inactivation can be fitted by the following modification of the h system of the Hodgkin-Huxley formalism: [Formula: see text]8. In the analysis it was not necessary to modify the concept of two separate processes, activation and inactivation, governing the opening and closing of the sodium channels.


On the role of barium in supporting the asynchronous release of acetylcholine quanta by motor nerve impulses. 1. The effect of Ba2+ on the evoked secretion of individual acetylcholine (ACh) quanta was studied on frog neuromuscular junctions using conventional electro-physiological techniques. 2. In solutions containing 1.8 mM-Ba2+ and no added Ca2+, 1 Hz stimulation for less than 1 min elevated miniature end-plate potential (m.e.p.p.) frequencies to 5-20 times the control level (seven experiments). Similar results were obtained when a Ca2+-chelating agent was added to the Ba2+ solution. 3. Repetitive nerve stimulation at frequencies greater than 1 Hz in concentrations of Ba2+ greater than or equal to 1.8 mM elevated m.e.p.p. frequencies to unmeasurable levels (greater than 100/sec). Such high m.e.p.p. frequencies were accompanied by a steady depolarization of the post-synaptic membrane, which was used to estimate the number of ACh quanta released. 4. The number of ACh quanta released asynchronously by nerve impulses was directly related to the external concentration of Ba2+ in a non-linear fashion. 5. Ba2+ was two orders of magnitude more effective than Ca2+ in supporting the evoked discharge of m.e.p.p.s. Ca2+ was a potent antagonist of asynchronous release in Ba2+ solutions. 6. Mg2+ and Co2+ both competitively antagonized evoked release in Ba2+ solutions. The equilibrium dissociation constant for each ion as an antagonist of asynchronous, Ba2+-dependent release was similar to its corresponding value as an antagonist of synchronous, Ca2+-mediated release. 7. It is suggested that Ba2+ supports dispersed, quantal ACh release directly by acting through the same conductance pathway normally traversed by Ca2+.


Effects of vascular perfusion on the accumulation, distribution and transfer of 3-O-methyl-D-glucose within and across the small intestine. 1. Factors affecting the transfer of the non-metabolized, ;actively transported' sugar, 3-O-methyl-D-glucose (3MG) across the small intestinal epithelium have been examined in vascularly perfused anuran intestine. Transfer has been studied during absorption in the steady state, and also during the period of transition from one steady state to another.2. During the steady state, the rate of absorption of 3MG from the intestinal lumen is equal to the rate of appearance in the portal venous effluent; this rate of transfer is to a small, but significant, extent directly related to the rate of arterial perfusion. With phlorizin in the intestinal lumen, transfer across the epithelium is reduced to very low rates which are independent of the rate of vascular perfusion.3. The apparent size of the tissue pool(s) of 3MG that have to be loaded to achieve the steady state rate of transfer are less than those that unload into the vascular bed after 3MG is removed from the intestinal lumen. This ;up-down asymmetry' is abolished when phlorizin is present in the intestinal lumen during the unloading phase.4. When 3MG is abruptly removed from the intestinal lumen after the tissue has been previously loaded with the sugar, the rate of washout into the vascular bed can be described by the sum of two exponential terms. The two terms differ in that the rate constant of the earlier ;fast' term is sensitive to the rate of vascular perfusion, while the later, ;slow', rate constant is insensitive to flow rate. The total quantity of 3MG that can be unloaded from the tissue into the portal venous effluent is decreased when phlorizin is present in the intestinal lumen during the unloading phase.5. Absorption from the lumen of the anuran intestine continues while the mesenteric circulation is interrupted. An estimate of the concentration of 3MG during the period of vascular stoppage can be made from the quantity recovered in the portal venous effluent when vascular perfusion is reinstituted (;vascular stop-flow'). The extent of the accumulation depends upon the duration of the vascular stoppage and the presence of Na ions in the intestinal lumen is essential for accumulation to occur.6. The findings are discussed in relation to the transfer of 3MG between various possible compartments in the tissue during absorption. Evidence is presented that a re-uptake of previously absorbed 3MG may occur across the brush border membrane. Such a recycling of 3MG across the epithelium implies that the apparent unidirectional fluxes measured across the epithelium between the bulk phase of the lumen and the blood may underestimate the size of fluxes across the epithelium at the cellular level.


The action of acetylcholine on background conductance in frog atrial trabeculae. The action of acetylcholine (ACh) on membrane potential and currents in frog atrial muscle has been studied with a double sucrose gap technique. The results show the following. 1. ACh induces the development of an extra current, outward at the resting potential, which is dependent on the ACh concentration. 2. The preparation does not show any sign of desensitization. 3. The reversal potential of the current induced by ACh is between 0 and 20 mV more negative than the resting potential and behaves as a K electrode. 4. The mechanism of ACh-induced K conductance presents inward going rectification properties. 5. The delayed outward current is not affected by ACh. However the evolution of its tail current seems to indicate a process of K accumulation related to the ACh-induced current.


Human thymus-leukemia-associated antigen: isolation and partial characterization. Human thymus-leukemia-associated antigen (HThy-L), a saline-soluble antigen, was previously detected by immunodiffusion (but not on the cell surface) in significant quantity in extracts of normal thymocytes, cells from cultured T-cell lines, and erythrocyte-rosette-positive leukemia blasts. Two species of HThy-L were identified and isolated from normal human thymus tissue after extraction in tris buffer, ammonium sulfate fractionation, acid precipitation of inactive fractions, DEAE-cellulose (DE-52) chromatography, Sephadex G-100 gel filtration, and carboxymethyl-cellulose (CM-52) chromatography; On Sephadex G-100, both HThy-L species had a similar molecular weight (40,000--50,000), but they eluted in different positions on DE-52 and CM-52. Analysis by sodium dodecyl sulfate polyacrylamide gel electrophoresis showed that each of the 2 HThy-L species contained 2 components with molecular weights of approximately 43,000  and 23,000. Further purification of HThy-L on Sephadex G-50 showed that the 43,000-dalton component possessed HThy-L activity.


Immunologic surveillance against chemically induced primary colon carcinoma in rats. A 1,2-dimethylhydrazine dihydrochloride-induced rat gastrointestinal tract tumor model was used to study the phenomenon of immunologic surveillance. In two different sets of experiments, a properly timed administration of antithymocyte globulin resulted in earlier tumor appearance, increased numbers of tumors, and increased multiplicity of gastrointestinal tumors. Results obtained from histologic examination of the gastrointestinal tract at different times after the last dose of 1,2-dimethylhydrazine dihydrochloride suggested that a normally functioning immune system effectively suppressed the growth of some nascent tumors. However, the immunosuppression of the host with antithymocyte globulin allowed the development of foci of microtumors into grossly visible neoplasms. Our experiments supported the concept that immunologic surveillance against neoplasia depends on the thymus cell system, although other possible mechanisms were not excluded.


Detection of metastatic tumors in nude mice: brief communication. Homozygous nude (nu/nu) mice were inoculated ip with either highly malignant human bladder transitional cell carcinoma or human prostate adenocarcinoma. These animals were subsequently given injections of normal human T-lymphocytes to restore the known T-lymphocyte deficiency present in homozygous nude mice. Metastatic spread of the prostate and bladder carcinomas was evident in mice given human T-lymphocytes. Although tumor growth was observed at the sites of tumor inoculation, no tumor spread was observed in mice not receiving T-lymphocytes.


Ibuprofen-induced meningitis in systemic lupus erythematosus. A patient with systemic lupus erythematosus had chills, fever, and headache on four separate occasions after ibuprofen ingestion. One such episode was accompanied by cerebrospinal fluid findings compatible with meningitis. Drug rechallenge under controlled conditions established the relationship between ibuprofen ingestion and meningitis.


Benefits from aortocoronary bypass graft. The first aortocoronary bypass graft (ACBG) for coronary heart disease (CHD) was performed in 1964. From 1967 to the summer of 1977, between 250,000 and 300,000 such procedures were performed in this country. With such extensive experience, the patient might expect that most questions regarding the effectiveness of the procedure in the treatment of CHD would have been answered. Unfortunately, this is not the case. After a decade of experience, all that is truly known about the procedure is that for most patients, it does improve the quality of life and, if present practices continue, will add greatly to the cost of medical care in this country. It appears that there is still a need for more randomized, controlled studies.


Activation of electrogenic Na+ pump by epinephrine in bullfrog atrium. The effects of epinephrine (1 X 10(-6)--1 X 10(-5) M) on the resting membrane potential and the K+-activated hyperpolarization of bullfrog atrial heart muscles were studied by the single sucrose-gap method or by intracellular microelectrode filled with 3 M KC1. Epinephrine-induced hyperpolarization (Ep-hyperpolarization) was completely eliminated by the effect of ouabain or by removal of extracellular K+. The amplitude of K+-activated hyperpolarization, which was produced when the extracellular K+ concentration was raised from zero to 2 mM, was markedly increased in the presence of epinephrine. The membrane depolarizations, due to high extracellular K+ concentration in the presence of ouabain (5 X 10(-6) M) which completely and reversibly eliminated the K+-activated hyperpolarization, were not altered under the effect of epinephrine. These results suggested that Ep-hyperpolarization of bullfrog atrial heart muscles was due to the acceleration of electrogenic Na+ pump which produced the K+-activated hyperpolarization.


Role of connectin in the length-tension relation of skeletal and cardiac muscles. Resting length-tension relation was studied in glycerinated preparations of frog semitendinosus and atrial muscles. The sarcomere length was measured by using either the light diffraction method or photomicrographs. When the glycerinated preparation was treated with 1 N NaOH solution and subsequently with 2% SDS solution, the resting tension decreased. In glycerinated semitendinosus muscle the resting tension decreased to below 20% of the value before treatment. However, the glycerinated atrial muscle retained more than 50% of its resting tension in spite of such biochemical treatment. The thermoelasticity of the muscle, which was represented by (formula:see text), was determined. Glycerinated semitendinosus and atrial muscles, which were treated with 0.6 M KCl, showed negative beta at all sarcomere lengths. However, when the glycerinated muscle was treated with both alkali (1 N NaOH) and SDS solution, beta at the shorter length became positive (i.e., a rubber-like property). Since connectin survived the biochemical treatment (either NaOH or SDS), it was concluded that the fraction of resting tension, which remained in spite of the biochemical treatments, might be due to the connectin.


Evaluation of thyroid functions in leprosy. III. Circulating auto-antibodies against thyroid and nuclear components. Sera from twenty six patients of various types of leprosy were tested for the detection of circulating auto-antibodies and nuclear components against thyroid using various methods. Four patients of lepromatous leprosy had higher levels of thyroid auto-antibodies by latex agglutination. Three patients showed the presence of anti-nuclear antibodies, two belonged to the TT and one to the LL group.


Optimal patency rates obtained in coronary artery grafting with circular vein grafts. Twenty-two patients underwent coronary artery grafting with a circular vein graft comprising four or five distal coronary anastomoses. Postoperative angiographic evaluation showed patency in 90 (95.7 percent) of the 94 coronary anastomoses fashioned in this manner. All four occlusions occurred in the one patient whose graft comprised five anastomoses and was occluded beyond the first anastomosis on the right coronary artery (RCA). Eleven of the 94 anastomoses were made onto arteries with limited runoff. Blod flow averaged 214 ml. per minute (range 130 to 320) in the main portion of the graft and 59 ml. per minute (range 35 to 100) in the most distal segment. Flow doubled (averaged 403 ml. per minute) in the 11 grafts injected with papaverine. The technique of circular vein grafting is described in detail and potential pitfalls are outlined. The reasons for the high early patency rate are believed to be the following: (1) high flow in the proximal segment of the graft insuring patency of said segment, (2) termination on the left anterior descending (LAD) coronary artery providing good distal flow and patency. (3) diamond-shaped side-to-side anastomoses (SSA's) preventing angulation of the graft at these crucial points, and (4) nearly equidistant anchoring at the site of the multiple anastomoses giving the graft a smooth, even contour.


Coagulation abnormalities in patients undergoing myocardial revascularization. One hundred patients were screened for hypercoagulability preoperatively and on the third, seventh, tenth, fourteenth, and twenty-first days postoperatively. Patients found to have hypercoagulability were treated with heparin, aspirin, and Coumadin. When the abnormality was present preoperatively, treatment was continued for the duration of the patient's life. Those patients in whom abnormalities developed postoperatively were given anticoagulants until cardiac catheterization 6 months following their operation. Twenty-four of the 100 patients had no coagulation abnormalities preoperatively or postoperatively. Fifteen patients were found to have abnormality prior to operation. Their predominant abnormality was low antithrombin III activity. Sixty-one patients became hypercoagulable postoperatively. Predominant abnormality in this group of patients was increased thrombin generation and increased platelet adhesiveness. Evaluation of patients in this study group revealed a decrease in the incidence of pulmonary embolism, an increase in the patency of vein grafts, and the elimination of anticoagulant therapy in 24 percent of the patients.


Early and late results of resection of ventricular aneurysm. Fifty-seven patients have undergone ventricular aneurysm resection with a mortality rate of 10 percent. Recent surgical trends have been toward complete revascularization and treatment of concomitant disorders when present. Although the early mortality rate may be favorable influenced by these maneuvers, analysis of survivors reveals no significant difference in survival between the group that had concomitant coronary artery bypass and the group that had aneurysm resection alone (84 percent versus 78 percent). There were significantly more asymptomatic patients in the revascularized group, however (74 percent versus 53 percent). Analysis of hospital deaths revealed nearly all to be seconary to low output syndrome, arrhythmia, myocardial infarction, or a combination of the three. Late death was due to either congestive heart failure or a myocardial infarction in all but one case. Seventy-seven percent of operative survivors are asymptomatic. Late morbidity is usually related to ungrafted or progressive coronary artery disease and generally has been successfully managed by reoperation.


Diaphragm pacing. Application to a patient with chronic obstructive pulmonary disease. In a 66-year-old patient with chronic obstructive pulmonary disease (COPD) complicated by arterial hypoxemia and repeated episodes of respiratory and right ventricular failure, a satisfactory level of oxygenation could not be maintained despite controlled oxygen therapy. To enable oxygen to be administered without depression ventilation, artificial respiration by means of phrenic nerve stimulation (diaphragm pacing) has been employed. Evidence of clinical improvement since pacing was begun 32 months ago include fewer episodes of respiratory failure and better control of congestive heart failure despite a gradual worsening of pulmonary function.


Ultrastructural studies of platelets of von Willebrand and normal swine. Platelets from normal pigs and pigs with von Willebrand's disease were studied by ultrastructural and scanning electron microscopy. The majority of resting platelets were discoid or abortive-dendritic in gross morphology with an average size of 2.6 micrometer. The number of alpha-granules per platelet section was varied; however, a highly significant positive association was found between platelet size and the number of alpha-granules. These studies characterize porcine platelets more definitively and clearly establish the similarities between platelets from normal and those from von Willebrand pigs.