[The postnatal development of lamina V pyramidal cells in the sensomotor cortex of the albino rat. 1. Introduction and qualitative studies of Golgi preparations]. 1. It is given an introductory survey about the usual Golgi procedures and modifications, and about the possible quantitative methods in Golgi researches. Upon them a synopsis of the literature about dendrite spine distribution and density in rodents' cortices (mouse, rat, hamster) follows. Concluding the preamble it is informed about the structural changes in spine distribution during postnatal ontogenesis published hitherto. 2. Using Golgi-Cox impregnated brain slices (according the RAMON-MOLINER modification) the differentiation of layer V pyramids in albino rat sensorimotor cortex during the 1st to the 90th day after birth is studied qualitatively. Several types of unknown interpyramidal synaptic contacts are published. Some notices follows about the first occurence of apical dendritic bundles during the postnatal ontogenesis, as well as about the axonal swellings of cortical pyramids.


[The postnatal development of lamina V pyramidal cells in the sensomotor cortex of the albino rat. 2. Quantitative studies on Golgi preparations]. Using Golgi-Cox preparates the postnatal development (1st to 90th day after birth) of the patterns of apical dendrite spines in layer V pyramids in albino rat sensorimotor cortex is represented. Especially it is reported about the evolution of the total amount of apical dendrite spines, of length of apical dendrites, of spine density (spine dendrite quotients, DQ), of interspinous distance (space between two adjacent visible spines, in the proximal dendrite with increasing spine distribution and in the district with crest of spine distribution separately) and finally of cell body length.


[Quantity of compentent cells in a population of Escherichia coli treated with Ca2+ cations]. An attempt was made to estimate the number of Escherichia coli K-12 cells rendered permeable to antibiotics under Ca2+ treatment. The effect of cold factor and Ca2+ alone as well as the cell age on the induction of permeability and the energy dependence of the latter were also investigated. About 70-75% and more exponentially growing cells as a result of Ca2+ treatment became sensitive to actinomycin, rubomycin and olivomycin. This number was somewhat lower (40-50%) in sationary phase culture. A fraction (20-30%) of stationary phase cells appeared to be sensitive to antibiotics even without Ca2+ pretreatment. Preincubation of the cells in cold in the absence of Ca2+ cations did not induce the cell permeability. The transport of antibiotics inside the cell was not prevented by an uncoupler of oxidative phosphorylation --carbonylcyanid-m-chlorophenylhydrazone (CCCP). It is suggested that the cells which are rendered permeable to tested antibiotics represent the "compentent" cells capable to uptake molecules of exogenous DNA as well.


Assessment of plasma fibrinolytic system with use of chromogenic substrate. A block of tests employing a chromogenic substrate H-D-Val-Leu-Lys-pNA (S-2251, Kabi AB, Stockholm) is proposed for a quantitative assessment of the plasma fibrinolytic system. It allows measurement of spontaneous plasmin activity, plasminogen content, activator activity (ability of plasma to activate plasminogen), immediate and progressive antiplasmin levels and alpha2-macroglobulin. All these assays could be performed within 45 min using a total of 0.4 ml of plasmin.


Alterations of plasmin activity, plasminogen levels and activity of antiplasmins during endotoxin shock in dogs. Spontaneous plasmin activity and 'fast-reacting' and 'time-dependent' antiplasmin activities were determined during various stages of canine endotoxin shock by means of assays utilizing a chromogenic tripeptide derivative (S-2251, Kabi AB, Peptide Research Division, Mölndal). During shock, gradually decreasing values of both antiplasmins were found, revealing most pronounced falls of 'fast-reactin' antiplasmin. These changes of plasma antiplasmin activities were accompanied by decreasing values of plasminogen and evidence of plasmin activity. Immunochemical determination of plasma levels of alpha2-macroglobulin (alpha2-M) and alpha1-antitrypsin (alpha1-AT) and gel filtration studies, demonstrated marked falls in levels of alpha2-M and alpha2-plasmin inhibitor and small reductions of alpha1-AT in this state.


[Factor XII (HAGEMAN FACTOR)].

[Factor XII (HAGEMAN FACTOR)]. The discovery of factor XII (Hageman factor), the attempts of characterizing it, the knowledge of its presence of absence in the various vertebrate classes are represented in a survey and recent opinions of molecular biology about its structure and composition are discussed. The effect of the activating substances for transferring the Hageman factor into its active form are represented with the role of the pre-kallikrein (Fletcher factor) and of the highly molecular kininogen (Fitzgerald factor) being referred to. Furthermore, the following systems influenced by factor XII are dealt with: 1. The endogenous and exogenous activating system of blood clotting and possible reasons for lower bleedings with factor XII deficiency. 2. Role of the Hageman factor in activating fibrinolysis. 3. Influence on the liberation of kinin. 4. Correlations towards the complement system. 5. Possible inhibitory effect of platelet aggregation. Finally the close connection of all these systems is referred to and the necessity of considering these complicated events in a complex way is stressed.


[Environmentally dependent erythrocyte membrane permeability as a source of error in the determination of hematocrit]. The volume of human red blood cells (RBC) was evaluated by means of the centrifugation method (hematocrit) and 131-J-labelled human serum albumin, respectively. Both of the methods yielded an identical volume of about 107 micron3 of the single RBC, provided the evaluation was performed in autologous plasma. Contrary to the 131-J-albumin method the results of which were found independent of various pretreatments of RBC, the centrifugation hematocrits of RBC previously washed with PBS and resuspended in PBS or saline protein media resulted in a mean cell volume of about 86 micron3. The decrease of the cell volume was associated with an efflux of K+ ions. If the RBC are centrifuged at 800 g instead of 15000 g, their volume will remain unchanged. The assessment of cytodeformability has shown, that RBC in PBS by loss of cell volume could enter a 2.3 micron micropipette completely. RBC in plasma, though traversing a 2.9 micron micropipette were incapable of entering a 2.3 micron channel completely. With pressures ranging from 300 to 350 mm H2O the processes of these cells undergo microspherulation.


[Extraction of membrane proteins in low ionic strengths]. Hemoglobin and the low molecular weight proteins 8 and 9 are extracted from ghosts during low ionic washing after the hypotonic hemolysis of erythrocytes. Furthermore, a loss of the proteins 4.5 and 7 was observed. The protein patterns of ghosts after isotonic hemolysis by freezing and thawing resemble the ghost protein patterns after hypotonic hemolysis and incomplete deprivation of Hb. Many if not all membrane proteins are eluted by repeated incubations of the ghosts in solutions of low ionic strength in the presence of EDTA. The spectrins, the proteins 5, 4.5, 7 and residual Hb are extracted preferentially. A selective extraction of the spectrins and the protein 5 is not detectable under these conditions. Often the spectrin bands are subdivided following low ionic incubation.


Coumarin-induced abnormal factor IX: an immunological study in humans. An excess of factor IX antigen or protein with respect to factor IX activity is present in coumarin treated patients. The average factor IX antigen value found in a group of 16 patients was 96.2 (S.D. = 24.46) whereas the average clotting activity was 19 (S.D. = 4.54) (p less than 0.001). In the electroimmunoassay system a normal peak or precipitate were seen in every instance. In haemophilia B--no peak or precipitate were seen. The coumarin induced abnormal factor IX shows a more anodic migration in the bidimensional immunoelectrophoresis system as compared with the normal counterpart. On the contrary, the factor IX protein present in haemophilia BM or in haemophilie B+ migrates as normal factor IX.


On the nature of fibrinolytic activity of arteries. The plasminogen activator of the arterial wall was studied with the histochemical method of TODD. The plasminogen activator was removed from the sections after extraction with M-potassium thiocyanate. By this procedure we suggest that the activator demonstrated by the histochemical method is the same substance as that prepared by the extraction method with thiocyanate of ASTRUP and STAGE. However, a new fibrinolytic activity was restored after treatment of these extracted sections with streptokinase or urokinase. There were no differences in the different types of arteries examined and normal or atherosclerotic arteries. Similar findings were found when kidney or myocardial tissues were examined. It is suggested that the arterial and other tissues contain proactivator-plasminogen which is not extracted from the tissue by potassium thiocyanate and can adsorb streptokinase or urokinase.


Studies on factor XIII antigen in congenital factor XIII deficiency. A tentative classification of the disease in two groups. Immunological and immunofluorescent studies carried out on plasma and platelets of three cases of congenital factor XIII deficiency are reported. Two of these patients were originally thought to have normal factor XIII subunit S and no subunit A. However, repeated assays carried out using different lots of antiserum showed that in reality the patients lacked both subunit S and subunit A. The false positive finding was due to the presence of a anti-factor VIII contaminant in the antiserum originally used. The third patient had a normal subunit S and no subunit A. No factor XIII antigen was found by the indirect immunofluorescent technique in normal, factor XIII deficiency and von Willebrand's disease platelets. On the contrary, by using the non-monospecific antiserum a fluorescent pattern similar to that observed by using an anti-factor VIII antiserum, had been noted. On the basis of the data presented in this paper a tentative classification of factor XIII deficiency in two groups is proposed: Type I is characterized by the lack of both factor XIII subunit S and A. Type II is characterized by a normal subunit S and no subunit A. The need for a re-evaluation of published case of factor XIII deficiency by means of monospecific antisera is indicated.


[The problem of preparing avid cytotoxic anti-B-lymphocyte serums]. There were anti-B-lymphocyte toxins in 40 samples (= 7.58%) from 527 examined sera of pregnant women and polytransfused persons without HLA cytotoxins and 84 samples (= 33.2%) from 253 anti-HLA sera. Of 49 specific anti-B sera 27 were suitable for typing B-lymphocyte antigens; with the help of 13 of these sera 5 specific antigens of B-lymphocytes called 1-5 could be determined. These 27 sera produced a positive cytotoxic reaction, mainly with the strength of +++ or ++. Attempts of absorbing HLA antibodies from anti-B lymphocyte sera led to unsatisfactory results. Sometimes the content of anti-B-lymphocyte toxins could also be diminished by a platelet absorption, in other cases the absorption was insufficient and had to be repeated therefore.


[Theoretical, methodological and clinical aspects of the growth of granulopoietic cells in agar culture]. Haematopoietic stem cells are capable of developing colonies of differentiated granulocytes in a semi-stable agar environment under corresponding experimental conditions. Number and size of aggregations developing from a single stem cell in each case called "in culture Colony Forming Unit (CFU-c)" enable conclusions to be made about the functional ability of the cultivated human or animal bone marrow. The method is applied in investigating the kinetics in the haematopoietic system and, in addition, it is used for diagnostics, therapy and control of the course in haematological diseases as well as to check the proliferous ability of cryopreserved bone marrow cells. As to the standardization of the procedure the instability of the active principle of the foetal calf serum as well as the essential colony stimulating factor represent limiting items at present.


[Fragmented erythrocytes (schizocytes) after surgical interventions]. Fragmented erythrocytes (schistocytes) develop as a consequence of erythrocytes being damaged mechanically. Cardiac and vessel anomalies as well as diseases of small vessels with disturbances of the microcirculation may be considered as the main causes. Intravasal haemolysis appearing after artificial heart valve replacement is characterized by the appearance of schistocytes. In laboratory chemical respect there are signs of a haemolysis. The treatment depends on the seriousness of clinical symptoms.


[Quantitative behavior of eosinophil granulocytes in the bone marrow of children with acute lymphoblastic leukemia]. Of 31 children affected with acute lymphoblastic leukaemia the quantitative behaviour of eosinophilie granulocytes was examined in the course of the disease. Nearly all patients were treated according to a chemotherapy scheme (Memphis IV). During this therapy the eosinophils greatly diminished initially increased significantly to subnormal values and to the values of healthy persons with persisting full remission. Another significant decrease occurred during the relapse and in the pre-final stage. During each following relapse a greater diminution of bone-marrow eosinophils could be observed. Simultaneously the decrease of eosinophils led to a shift in the degree of maturation. In this connection the similar behaviour of neutrophilic and eosinophilic granulocytes of the bone-marrow must be stressed. Eventually, the lbast excrescence in the bone-marrow and its therapy cannot solely be decisive for the findings made. Relations to the lymphocytic system can be referred to.


N-acetyl-neuraminic acid (NANA) serum level in the diagnostics of preleukemic states, acute micromyeloblastic leukemias and pancytopenias. The levels of N-acetyl-neuraminic acid were determined in patients with preleukemic states, acute micromyeloblastic leukemias and pancytopenias. A statistically significant increase of NANA was found in patients with micromyeloblastic leukemia in comparison with preleukemic states and pancytopenias. A significant rise in the NANA level was observed in preleukemic states in comparison with pancytopenia of other origins. The assay of the NANA level may be employed as a sensitive biochemical test for differential diagnostics of these diseases.


[The karyotype of bone marrow cells in the preleukemic phase of chronic myelocytic leukemia]. The caryotype of bone-marrow cells of 19 patients affected with leukocytosis of obscure origin was examined by means of G-staining. A chronic myelogenous leukaemia in its preleukaemic stage was diagnosed in 11 patients by identifying Ph'-chromosomes in all metaphases observed and a lowered acid leukocyte phosphatase. Besides Ph'-chromosomes an aneuploida (47 chromosomes with trisomia 8) was observed in one patient. The diagnostic and prognostic significance of this pathological caryotype is discussed.


[Vitality studies in bone marrow cells by means of fluorescence microscopy]. In assessing the vitality of bone-marrow cells vital fluorochroming with acridine orange allows a differential, qualitative statement to be made about the degree of cell damage and in addition the single cell compartments to be relatively well differentiated in morphological respect. The stain exclusion test made for the purpose of comparison merely enables an approximate, quantitative evaluation to be obtained.


[Hematologic complications in acute alcoholism]. The authors investigated the relations between cell anomalies (vacuolation, increasing sideroblastosis) caused by the uptake of alcohol and the dynamics of haematopoesis in the bone-marrow of 33 alcoholists who had been admitted in a comatose condition and who were neither affected with anaemia nor with chronic hepatitis. In all cases a maturation arrest of erythropoetic and granulopoetic cell elements which was not in accordance with the number of immature vacuolated cells could be observed. 12 test persons showed increased sideroblastic indices and a slightly diminished medullary reticulocytosis. The majority showed a very active thrombocytopoesis contrasting with the normal or even diminished number of thrombocytes in the peripheral blood. The authors come to the conclusion that alcohol will cause a general metabolic damage of haematopoesis and at the same time it will produce a direct toxic effect on the bone-marrow cells (proerythroblasts, promyelocytes) and the peripheral blood (thrombocytes).


A clinicopathological study of non-Hodgkin's malignant lymphomas in terms of the Kiel classification. A group of 25 patients with non-Hodgkin's lymphoma was investigated. Patients diagnosed earlier were reclassified according to the Kiel system. A correlation between age distribution and histological malignancy was found. The time between the onset of symptoms and diagnosis was 1 year on an average. The majority of the patients belonged to stage IV. The survival rate was higher in the low-grade malignancy group than in the high-grade group. When assessing the prognosis, the histological classification as well as the clinical staging ought to be considered. The bone marrow was the most frequent extranodal site of involvement in stage IV. Cytopenic changes were almost invariably accompanied by bone marrow infiltration. All the 7 cases analysed for lymphocyte surface markers proved to be B cell type. No significant difference was seen between the results of single agent and combined chemotherapy.


The iron status in healthy individuals aging from 18-25 years. The haemoglobin, haematocrit, erythrocyte count, transferrin and serum iron values of a group of 65 healthy young people aging from 18-25 years were determined. Ferritin in serum was quantitated by radioimmunoassay to determine the usefulness of this assay in reflecting iron stores of healthy people.


[The question of the diagnostic value of increased erythrocyte enzymes in leukemia]. The investigated erythrocyte enzymes (LDH, pyruvatkinase, Glukose-6-phosphat-dehydrogenase) are moderately to extremely elevated in most cases of leucemia. There exists no significant correlation to the number of reticulocytes and the erythrocyte half life time.


[Anti-allergic activity of 7-acetyl-5-oxo-5H-[1] benzopyrano (2,3-b] pyridine (Y-9000) (author's transl)]. The IgE mediated reactions such as 48 hr homologous passive cutaneous anaphylaxis (PCA) and active anaphylactic bronchoconstriction in rats were inhibited in a dose dependent manner by treatment with 7-acetyl-5-oxo-5H-[1]benzopyranol[2,3-b] pyridine (Y-9000) and disodium cromoglycate (DSCG) given intraperitoneally. The inhibitory activity of Y-9000 was to the same extent as that seen with DSCG. The IgE mediated reactions were also inhibited by oral treatment with Y-9000 but not with DSCG. In addition, the treatment with Y-9000 resulted in inhibition of IgG mediated reaction such as anaphylactic asthma in the passively sensitized guinea pigs and 4 hr heterologous PCA in rats. However, DSCG failed to prevent these reactions. Y-9000 also inhibited the active systemic anaphylaxis of the mouse and non-immunological reactions in rats such as histamine release after an intraperitoneal injection of dextran, anaphylactoid reaction and paw edema induced by the dextran, egg white or carrageenin. This agent had a stimulating effect on the adrenals, and showed glucocorticoid like activity, but bronchodilator and antagonistic activities on chemical mediators were nil. These results suggest that the anti-allergic activities of Y-9000 are elicited by inhibiting the release of allergic mediators in a manner similar to DSCG, and are partially mediated by stressor activity.


[Studies on D-penicillamine (1): Inhibitory effect of D-penicillamine on the heat-Cu++ induced denaturation of human gamma-globulin (author's transl)]. The inhibitory effect of D-penicillamine on the denaturation of human gamma-globulin induced by heat and Cu++ was compared with the action of other agents such as antirheumatic drugs, anti-inflammatory drugs, SH reagents, SH inhibitors and chelating reagents. The denaturation of human gamma globulin was induced by Cu++ at 10 micrometer ("Cu++ induced denaturation") and was further increased by heating at 63 degrees C for 3 hr in the presence of Cu++ ("total denaturation"). Thus the value obtained by subtracting "Cu++ induced denaturation" from "total one" was designated as "thermal denaturation". D-Penicillamine enhanced "thermal denaturation" at a low concentration but inhibited it with increasing the concentration as well as L-cysteine. SH reagents such as thiomalic acid, 6-mercaptopurine inhibited "total" and "thermal" denaturation. SH inhibitors and protein binding reagents such as N-ethylmaleimide, trinitrobenzenesulfonic acid inhibited the "total" and "thermal" denaturation of the protein. Chelating reagents such as ethylenediamine tetraacetic acid, 8-hydroxyquinoline inhibited "total", "Cu++ induced" and "thermal" denaturation of the protein. Au+ inhibited "total denaturation", but not "Cu++ induced denaturation". On the other hand, Au+++ denaturated the protein considerably with or without heating, in the absence of Cu++ but dithiothreitol did so only with heating in the same condition. The anti-inflammatory drugs used herein had no effect on the protein denaturation. D-Penicillamine apparently prevents the denaturation of human gamma-globulin by the chelate formation with Cu++ and the binding to free protein SH, initiator for sulfhydryldisulfide interchange reaction.


The occurrence of hyaline cytoplasmic inclusions in the nerve cells of the hypoglossal nucleus in human autopsy material. Among 440 adults (ranging in age from 41 to 99 years) who were examined for hyaline cytoplasmic inclusions (HCI) within cells of the hypoglossal nuclei of the medulla oblongata, HCI were found in 75 (17.0%). No definite correlation was noted between HCI and specific diseases. The morphological and histochemical descriptions and pathological interpretation as well as frequency of HCI as reported in the literature were reviewed. There was no evidence that the presence of HCI was related to a specific disease, ages or sex.


Serum antiproteases and respiratory diseases of the horse. The serum antiprotease (AAT) levels are reported in healthy horses and horses with respiratory diseases. Of the methods used, only the STIC test seemed to give useful results; this test showed variations in horses with respiratory diseases, especially in horses with acute alveolar pulmonary emphysema.


Suppressor T cells and suppressor factor. The ability to suppress antibody response to specific antigens has been found to be inherent in a subpopulation of T lymphocytes. The suppressor pathway has been shown to involve an initial T-cell clone that that elaborates a soluble factor which, in turn, induces a second cohort of effector cells. The implications of this system for autoimmune and neoplastic diseases--and some relevant experiments--are discussed.


Antiserum to surface antigens as a marker for cultured rat lung endothelial cells (1). Antibodies were developed in rabbits against an established line of endothelial cells from normal adult rat lung. Pre- and postimmunization sera were tested for antibody activity by the indirect immunofluorescence technique. Preimmunization serum failed to react with the endothelial cells, whereas the antibody titer of postimmunization serum from two rabbits was 1:512. Organ specificity and species specificity were assessed by absorbing the serum with packed dissociated cells from different organs of the rat and lung cells of other species. Only cells obtained from rat lung absorbed the antibodies completely. The antiserum showed some crossreactivity with the other cultured cells but the pattern of fluorescence was different. In the presence of complement the antiserum was found to be cytotoxic to cultured rat lung endothelial cells but not to the other crossreacting cells.


Target cell - substratum interaction. I. Effect of primed lymphocytes on a rat mammary adenocarcinoma tumor cell line. The differential effects of normal and immunologically primed lymphocytes on the adherence of trypsinized rat mammary adenocarcinoma tumor cells (DMBA"8) to their substratum are described, utilizing both allogeneically and syngeneically primed lymphoid cells as effectors. This observation may illustrate an important immunological phenomenon, and may result in a simplified assay procedure for the detection of cell-mediated immunity.


Specificity of murine delayed-type hypersensitivity to conjugates of large or small haptens on protein carriers bearing lipid groups. Delayed-type hypersensitivity (DH) in the mouse was provoked with different hapten-carrier complexes mixed with the cationic, surface-active lipid, dimethyl dioctadecyl ammonium bromide (DDA). DH was measured as footpad swelling. Conjugates of bovine serum albumin (BSA) with the small haptens dinitrophenyl (DNP), 'arsonate' (ARS) and 'sulphonate' (SULPH) served to generate strong DH reactions towards the homologous antigen. Insertion of a tripeptide spacer between the hapten and carrier resulted in lower DH reactivity. Optimal dosages and optimal time intervals between sensitization and DH elicitation were determined for the enlarged hapten-carrier complexes. Cyclophosphamide (CY) treatment, before priming with complexes mixed with DDA, caused a 5-6 day delay in the expression of DH but failed to evoke enhanced DH for any of the antigens tested. A broad array of cross reactions between small and enlarged hapten-carrier complexes showed a relative lack of specificity in these DH responses. The results are compared with others reported in the literature and are explained mainly by the effects of electrostatically bound lipid groups of DDA in the sensitizing conjugates.


Reaction mechanisms of beta1H globulin.

Reaction mechanisms of beta1H globulin. The reaction mechanisms of beta1H were studied. The generation of alternative pathway C3 and C5 convertases on the cell surface as well as in the fluid phase was inhibited by beta1H globulin. The cell preparation bearing the C3b site could bind beta1H with little effect on the C3b hemolytic activity. Bound beta1H was dissociated by the action of C3bINA and C3bINA-treated C3b bearing cell did not bind beta1H anymore. Cell-bound beta1H was also dissociated by the action of B (or Bb). From these and other results, the following conclusions were obtained. The C3b site-bearing cell could bind beta1H on the C3c region of C3b molecules facilitating the C3bINA action on C3b, and beta1H shared the same binding site with B (or Bb) inhibiting the generation of the alternative pathway convertases competitively.


Delay in the development of the allergic response to metals following intratracheal instillation. Intracheal intubation with the soluble metal salts potassium dichromate (K2Cr2O7) and nickel sulphate (NiSO4) causes a delay of up to 8 weeks in the development of delayed hypersensitivity to the specific agent. It is suggested that absorption of sensitizers by the respiratory route may, under certain circumstances, induce a state of specific immunological unresponsiveness, rather than necessarily lead to the development of a state of allergic sensitivity.


Proteolytic release of antigenic fragments corresponding to normal fecal antigen and non-specific cross-reacting antigen from carcinoembryonic antigen. Three immunogenic parts have so far been identified in the carcinoembryonic antigen (CEA) molecule. These are: determinants cross-reactint with the normal fecal antigen (NFA) (NFA determinant); determinants cross-reacting antigen (NCA) (NCA determinant); and determinants which appear to be more cancer-specific (cancer determinant). The chemical nature of these parts of the CEA molecule was investigated by digestion with proteolytic enzymes together with anti-CEA preparations with which these three immunogenic parts of CEA molecule could be identified. The CEA digest obtained with pepsin did not react in immunodiffusion and radioimmunoassay, indicating that pepsin completely destroyed all the antigenic parts. Digestion by pronase E destroyed only the cancer determinant and liberated two antigenic fragments corresponding to the NFA determinant and the NCA determinant, respectively. These results suggest that the cancer determinant may reside in a protein or a peptide part of the molecule. The chemical nature of the NFA and NCA determinant remains to be clarified.


Orientation of acidic polysaccharides and rhodopsin-oligosaccharides in frog retinal rod outer segments. Using topo-optical staining reactions, the presence and molecular order of three structural components of outer segments of frog retina were studied. These components included (1) an acidic polysaccharide texture, (2) free aldehyde groups which arise during formalin fixation and (3) the oligosaccharide chains of rhodopsin. Quantitative measurements of the dye binding and birefringence effects arising from the individual structural components in rod outer segments were made. Results indicated that all three structural components had a rather well-defined orientation within the ROS. The spherulites phagocytized from the apical ends of ROSs by the pigment epithelium also demonstrate preferred orientation of the three structural components investigated.


Purification and partial characterization of the B subunit of Serratia marcescens tryptophan synthetase. A trpE mutant of Serratia marcescens (E-7) was isolated, and the multimeric enzyme tryptophan synthetase (EC 4.2.1.20) was purified to homogeneity from derepressed cells. The A and B subunits were resolved, and the B subunit was partially characterized and compared with the Escherichia coli B subunit as part of a comparative evolution study of the trpB cistron of the trp operon in the Enterobacteriaceae. The S. marcescens B subunit is a dimer (beta(2)), and its molecular weight was estimated to be 89,000. The separate subunits (beta monomers) had molecular weights of approximately 43,000. The B subunit required pyridoxal phosphate for catalytic activity and had an apparent K(m) of 9 x 10(-6) M. The N terminus of the B subunit was unavailable for reaction with terminal amine reagents (blocked), whereas carboxypeptidase digestion released a C-terminal isoleucine. Using S. marcescens B antiserum in agar immunodiffusion gave an almost complete reaction of identity between the B subunits of S. marcescens and E. coli. The antiserum was used in microcomplement fixation, allowing for a comparison of the overall antigenic surface structure of the two B subunits. The index of dissimilarity for the heterologous E. coli enzyme compared with the homologous S. marcescens enzyme was 2.4, indicating extensive similarity of the two proteins at their surfaces. Comparative antiserum neutralization of B-subunit enzyme activity showed the E. coli enzyme to cross-react 85% as well as the S. marcescens enzyme. With regard to the biochemical and immunochemical parameters used in this study, the S. marcescens and E. coli B subunits were either identical or very similar. These findings support the idea that the trpB cistron of the trp operon is a relatively conserved gene in the Enterobacteriaceae.


Detection, quantitation and contrast enhancement of myocardial infarction utilizing computerized axial tomography: comparison with histochemical staining and 99mTc-pyrophosphate imaging. The potential role of the CAT scan in detecting and quantitating acute myocardial infarctions was assessed in the excised hearts of 25 dogs with 48-hour-old myocardial infarctions. The CAT scan detected the infarct in every instance and distinguished subendocardial from transmural infarctions. While the CAT scan slightly underestimated the volume of infarcted tissue, this volume had a close linear relationship to the true volume determined by histochemical staining. On the other hand, the 99mTc-pyrophosphate scintiscans grossly overestimated the volume of infarcted tissue. Intravenous administration of contrast material prior to sacrifice produced contrast enhancement either of the rim or diffusely throughout the infarct. Thus CAT should have an important role not only in detecting but also in estimating the volume of acute myocardial infarctions.


Enzyme-bound formylvaline and formylvalylglycine; an initiation complex for gramicidin A biosynthesis. Gramicidin A is an antibiotic peptide produced by Bacillus brevis ATCC 8185, which also produces tyrocidines. An attempt was made to establish a cell-free enzyme system for gramicidin A synthesis. An enzyme fraction, Component I, was partially purified from crude extracts of the organism and proven to be involved in the synthesis of the formyl-Val-Gly- region of gramicidin A. The initiation of gramicidin A biosynthesis is a function of Component I, which activates valine and binds it as a thioester, and further formylates it in the presence of formyltetrahydrofolic acid. The formylvaline thus synthesized is transferred to the glycine moiety, which is also thioesterified to Component I. Elongation of the peptide chain takes place by a mechanism similar to those found for tyrocidines, gramicidin S, and bacitracin.


Structure and orientation of cytochrome c oxidase in crystalline membranes. Studies by electron microscopy and by labeling with subunit-specific antibodies. The structure and the orientation of cytochrome c oxidase molecules in crystalline cytochrome c oxidase membranes (Vanderkooi, G., Senior, A.E., Capaldi, R.A., and Hayashi, H. (1972) Biochim. Biophys. Acta 274, 38-48) were studied by image analysis of electron micrographs and by reacting the crystalline preparations with immune gamma-globulins against individual cytochrome c oxidase subunits. Binding of gamma-globulins to the membranes was detected by the following two methods: (a) electrophoretic identification of gamma-globulin polypeptides in the washed membranes; (b) electron microscopic examination of the negatively stained membranes. The membranes bound immune gamma-globulins against subunit IV (which faces the matrix side in intact mitochondria) but failed to bind immune gamma-globulins against subunits II + III (which face the outer side of the inner membrane in intact mitochondria). In contrast, solubilized cytochrome c oxidase bound either of the two immune gamma-globulins. All cytochrome c oxidase molecules in the crystalline membranes are thus asymmetrically arranged so that subunit IV faces outward and subunits II + III face toward the interior. This orientation is opposite to that found with intact mitochondria. The data also suggest that the crystalline membranes form closed vesicles which are impermeable to externally added gamma-globulins.


Radiologic, clinical and biochemical features of non-seminomatous testicular tumors. Current therapy for metastatic non-seminomatous testis tumors is effective but toxic. Careful investigation is needed to select those who need treatment and yet avoid needless toxicity in patients who have no disease. Current radiographic investigations and measurement of two biochemical tumor markers (AFP & BHCG) provided accurate monitoring in 19 patients. Preoperative urography and lymphography correctly predicted the presence or absence of retroperitoneal metastases in 9 of 12 patients but did not show microsocpic metastases in 3. There was good correlation between the clinical, readiographic and biochemical evidence of disease progression and regression. Serial ultrasonography and tumor marker determinations were particularly useful in following asymptomatic abdominal metastases. Radiography and tumor marker determinations each have specific advantages which make them complementary.


Electron microscope studies of pH effects on assembly of tubulin free of associated proteins. Delineation of substructure by tannic acid staining. Bovine brain tubulin, purified by phosphocellulose chromatography (PC), was assembled in the presence of 10% dimethyl sulfoxide (DMSO), and the reaction was monitored turbidimetrically. Samples were fixed in glutaraldehyde-tannic acid after completion of polymerization, as indicated by no further change in absorbance, and then sectioned and studied electron microscopy, with special attention being given to the arrangement of protofilaments in the walls of formed elements. Samples of PC-tubulin were polymerized in buffer having various pH values from 6.0 to 7.7. At the lower pH values, only branched and flattened ribbons of protofilaments are formed. At intermediate values, the ribbons are unbranched, narrower, and more curved in cross section; complete microtubules are also seen. At the higher pH values, the predominate formed elements are complete microtubules. Most of the complete microtubules examined in this study had 14 wall protofilaments. The effect of pH on tubulin assembly was shown not to be an effect of DMSO. The dimers of associated protofilaments in ribbons and microtubules are conceptually viewed as having trapezoidal profiles in cross section, and, as additional dimers are added, the "C"-shaped ribbon closes to form a tube. The tilt angle of the lateral surfaces of the "trapezoidal" dimers will determine the number of wall protofilaments in the microtubules. At low pH, it is theorized that the trapezoidal profile of the dimer is shifted to a more rectangular configuration such that flat ribbons are formed by the lateral association of dimers. Also, variously shaped ribbon structures are formed at intermediate pH values, including "S"- and "W"-shaped structures, and elements shaped like a figure "6," all representing ribbons viewed in cross section. By visualizing the trapezoidal dimer in three-dimensions, and by arbitrarily indexing its six binding surfaces, it is possible to discuss interdimer binding in terms of preferred and possible binding interactions.


Degradation products of fibrinogen by elastase-like neutral protease from human granulocytes. Characterization and effects on blood coagulation in vitro. We investigated the effect of elastase-like neutral protease isolated from human granolocytes on human fibrinogen. Dependent on enzyme concentration and time of incubation, the elastase-like protease induced a progressive degradation of fibrinogen. Analysis of the remaining polypeptide chains showed a high susceptibility of the Aalpha- and low susceptibility of the gamma-chain of fibrinogen towards the proteolytic action of the enzyme. The split products were characterized by polyacrylamide gel electrophoresis and two-dimensional immunoelectrophoresis. They showed antigenic determinants of fibrinogen and of plasmin-induced proteolysis products D and E. The cleavage fragments isolated by gel chromatography had distinct molecular weights. Coagulability of fibrinogen by thrombin was inhibited according to the concentration of the protease and the time of incubation. Split products of fibrinogen with higher molecular weight prolonged the coagulation time of native fibrinogen, whereas low molecular weight fragments were ineffective.


Serotonin neurons of the midbrain raphe: ascending projections. The ascending projections of serotonin neurons of the midbrain raphe were analyzed in the rat using the autoradiographic tracing method. Axons of raphe serotonin neurons ascend in the ventral tegmental area and enter the medial forebrain bundle. A number of fibers leave the major group to ascend along the fasciculus retroflexus. Some fibers enter the habenula but the majority turn rostrally in the internal medullary lamina of the thalamus to innervate dorsal thalamus. Two additional large projections leave the medial forebrain bundle in the hypothalamus; the ansa peduncularis-ventral amygdaloid bundle system turns laterally through the internal capsule into the striatal complex, amygdala and the external capsule to reach lateral and posterior cortex, and another system of fibers turns medially to innervate medial hypothalamus and median eminence and form a contrelateral projection via the supraoptic commissures. Rostrally the major group in the medial forebrain bundle divides into several components: fibers entering the stria medullaris to terminate in thalamus; fibers entering the stria terminalis to terminate in the amygdala; fibers traversing the fornix to the hippocampus; fibers running through septum to enter the cingulum and terminate in dorsal and medial cortex and in hippocampus; fibers entering the external capsule to innervate rostral and lateral cortex; and fibers continuing forward in the medial olfactory stria to terminate in the anterior olfactory nucleus and olfactory bulb.


Intravenous gamma globulin in the management of patients with hypogammaglobulinemia. The safety and efficacy of plasmin-treated gamma globulin (PG-GG), an intravenous preparation with low anticomplementary activity, was assessed as an antibody replacement therapy in 14 patients with hypogammaglobulinemia. Seven were studied in 2 treatment periods of 6 and 15 mo on PT-GG with an intervening control period of intramuscular gamma globulin (IM-ISG). Frequency of infusions ranged from 2 to 4 wk to maintain a serum IgG concentration of less than 2.5 mg/ml. Three patients with severe chronic pulmonary disease were removed from the study because of lack of clinical improvement and were placed on single-donor plasma. The remaining 11 patients had a decrease in the number of hospitalizations or severe infections. Five patients had one or more systemic reactions (21/240 infusions). Symptoms abated rapidly with temporary interruption of the infusion. From these results, we conclude that PT-GG represents a relatively safe, efficacious mode of replacement therapy which has had uniformly high acceptance in patients with hypogammaglobulinemia.


Fluorescence properties of free and protein bound fluorescein dyes. I. Macrospectrofluorometric measurements. Excitation and emission properties of fluorescein derivatives were studied macrofluorometrically. Measurements were performed with solutions of various concentrations (0.07-100 microgram/ml) of free sodium fluorescein prepared from fluorescein diacetate (FDA), fluorescein isothiocyanate (FITC) and FITC bound to rabbit gamma-globulin. Both excitation and emission spectra as well as fluorescence intensities at constant excitation/emission wavelengths (496/515 nm) were recorded. The findings indicate that (1) FDA gives about twice the fluorescence intensity compared to equal concentrations of FITC. (2) The fluorescence properties of FITC upon excitation with blue light (lambda = 496 nm) are only slightly altered by the conjugation to rabbit gamma-globulin. (3) Considerable quenching due to conjugation could, however, be shown to occur upon UV excitation (lambda = 340 nm). (4) Fluorescence emission excited by visible blue light (496 nm) increases linearly to dye concentration in a range of 0.07-2.5 microgram/ml. Beginning at 5 microgram/ml (10-(5) M/1) all three compounds show a sharp decrease of fluorescence intensity with further increasing concentration. Practical aspects of these data for the immunofluorescence method are discussed.


A fixation method for improved antibody penetration in electron microscopical immunoperoxidase studies. A fixation method for electron microscopical immunoperoxidase staining has been developed, which (a) allows penetration of antibodies through cell membranes to intracellular antigen sites, (b) provides a reasonable cell preservation and (c) does not alter the antigenic structure in too great an extent. Penetration of the antibodies has been achieved by using saponin as a cell membrane attacking agent. The best results could be obtained after pretreatment of cell monolayers with a mixture of 0.05% saponin, 0.0125%-0.05% glutaraldehyde and 1% paraformaldehyde for 5 min at 4 degrees C, and postfixing them with the corresponding fixative without saponin for 45 min at 4 degrees C.


Ultrastructural localization of basic proteins in Trypanosoma cruzi. The postformalin ammoniacal silver (AS) and the ethanolic phosphotungstic acid (EPTA) techniques were applied in epimastigote and trypomastigote forms of the pathogenic protozoa Trypanosoma cruzi to detect basic proteins at the ultrastructural level. With both techniques, reaction was observed in the nucleus and in some cytoplasmic vacuoles. In the kinetoplast of epimastigotes, reaction was observed only at its periphery. In trypomastigotes, however, an intense reaction was observed in the spherical kinetoplast. With the ethanolic phosphotungstic acid technique, reaction was also observed in ribosomes and at the peripheral doublet microtubules of the flagellum. The filaments which form the paraflagellar structure did not react.


Use of hydrogen peroxide-egg albumin to eliminate nonspecific staining in immunoperoxidase techniques. The application of hydrogen peroxide and egg albumin to paraffin sections before immunostaining prevents nonspecific staining by immunoperoxidase techniques. This method is more effective than pretreating secions with normal sera, or using either egg albumin or hydrogen peroxide separately, or using diluted antisera with prolonged incuabations in the staining procedure.


Negative staining of whole cells: transmission electron microscopy of peripheral organelles in rat venous endothelial cells. The study of whole negatively stained cells has revealed details of cellular organelles in rat venous endothelial cells. In particular, details of surface membrane organelles and small tubular structures were demonstrated. The surface membrane organelles which appeared "vesicular-like" were found to be connected with small tubular attachments. These findings were correlated with those described by other techniques. It is significant that this simple technique appears to permit the demonstration of fine details of three-dimensional cytoplasmic structures.


Solubilization and partial characterization of a tumor-associated transplantation antigen of the guinea pig L2C leukemia. A tumor-associated transplantation antigen (TATA) from guinea pig L2C leukemia cells was solubilized by different methods. It was found that the 3 M KCl extraction yielded the most immunogenic TATA of L2C cells. Immunization of normal strain 2 guinea pigs with this extract in complete Freund's adjuvant gave complete protection against a subsequent challenge with tumor cells. Further fractionation of the KCl extract of L2C cells by Sephadex G-200 chromatography suggested that the immunogenic activity was present in the fraction containing materials with estimated m.w. of less than 20,000 daltons.


Idiotype-specific transplantation resistance to MOPC-315: abrogation by post-immunization thymectomy. The idiotype-specific myeloma transplantation resistance induced in BALB/c mice by immunization with the DNP-binding IgAlambda2 protein produced by plasmacytoma MOPC-315 is ablated by post-immunization thymectomy. Sham-thymectomy has no effect. The ablative effect of thymectomy is observed is observed in mice challenged subcutaneously with MOPC-315 cells either 3 days after thymectomy, or after a rest period of 44 days after thymectomy. These observations suggest that short-lived, thymic-dependent suppressive factors may play a role in the idiotype-specific myeloma graft resistance.


Murine Ia antigens: identification and mapping of Ia.23 and further definition of the I-E subregion. The Ia.23 specificity is a private specificity of the H-2d haplotype which we have assigned to the I-E subregion. It is found on the same Ia molecule that bears Ia.7, a public specificity previously assigned to the I-C subregion of haplotypes H-2d, H-2k, H-2p and H-2r. We suggest a reassignment of specificity Ia.7 to the I-E subregion to simplify the interpretation of the data presented in this paper and elsewhere, and to permit an orderly description of that part of the I region to the right of I-J.


Hapten-specific tolerance induced by hapten conjugates of D-glutamic acid, D-lysine (D-Gl) or isologous gamma-globulin: evidence for central B cell tolerance in the presence of carrier-primed helper T cells. A comparison has been made of the well known hapten-specific tolerance systems induced, respectively, by hapten-D-GL or hapten-isologous gamma-globulin conjugates. The principal question addressed in this study concerned the comparative maintenance of B cell tolerance, induced by one or the other method, after adoptive transfer into carrier-primed, irradiated recipient animals and, in addition, what role, if any, might be played by T lymphocytes in the tolerant donor cell population in maintaining such tolerance. The results clearly show that insofar as the hapten-specific B cell is concerned, no obvious difference exists in the capacity to maintain tolerance adoptive transfer between the hapten-D-GL and hapten-isologous gamma-globulin systems; such cells remained tolerant even in the presence of excess helper T cell activity. Moreover, under the conditions employed, depletion of T lymphocytes from the tolerant donor cell population did not affect the maintenance of hapten-specific B cell tolerance after adoptive transfer to irradiated recipients.


Alloantigens determined by a second D region locus elicit a strong in vitro cytotoxic response. The newly defined L antigen (previously designated D') has been studied with the CML assay. Balb/c-H-2db mice, which carry a loss mutation in the D region, were found to generate anti-L cytotoxic effector cells when stimulated with wild-type (BALB/c) cells. When the target cells were treated with an antiserum against L, cytolysis by BALB/c-H-2db anti-BALB/c effectors was completely blocked. However, an antiserum against D, the antigen that bears the D region private specificity, had no effect on the anti-L cytotoxic response. In a control CML experiment that was specific for the D antigen, the antiserum to L had no effect, whereas the antiserum to D blocked completely. These results indicate that the L and D antigens are recognized in a CML response as separate entities on the cell surface. In an allogeneic response to the products of the D region, cytotoxic effectors were generated that were specific for L in addition to those specific for D. This conclusion was supported by blocking experiments with either antisera or unlabeled targets. These functional studies thus extend previous co-capping and immunochemical studies, and further support the hypothesis that L is a third transplantation antigen similar to the K and D antigens.


Leukocyte-adherence inhibition: a specific assay of cell-mediated immunity dependent on lymphokine-mediated collaboration between T lymphocytes. The leukocyte-adherence inhibition (LAI) assay was studied to determine its immunologic relevance and identify the cell populations on which it depends. Two systems were employed: peripheral blood leukocytes from humans immunized with KLH, and lymph node cells from rats immunized with DNP-BCG. In both cases, LAI responses appeared about 3 to 4 days after immunization, reached a peak about 3 to 4 weeks later, and diminished thereafter. Reimmunization resulted in a booster-like response. LAI analysis in both systems showed dose-response dependency. Responses could be elicited only with the immunizing antigen. Virtual depletion of phagocytic cells had no effect on the response. E-rosette-forming cells gave an excellent response to KLH and also produced an active supernatant (lymphokine). Cells not forming spontaneous E-rosettes were inactive and could not produce active supernatants. Only those nonimmune cells that formed E-rosettes could respond to active supernatants. Thus, the LAI response is a specific indicator of cell-mediated immunity. T lymphocytes probably are required both at the antigen-reactive stage and at the stage of responding to the T cell-dependent lymphokine.


Adherent cell function in murine T lymphocyte antigen recognition. I. A. macrophage-dependent T cell proliferation assay in the mouse. The data in this report describe a T cell proliferation assay with nylon wool column-purified murine lymph node lymphocyte from animals immunized by footpad injection of antigen in CFA. It was found that the in vitro immune response of sensitized T cells to soluble protein antigens was functionally dependent on the presence of adherent cells, more specifically macrophages, at all concentrations of in vitro antigen challenge. The response was due to T cells in that cytotoxic treatment of the immune lymphocyte cells with anti-Thy 1.2 serum and complement effectively eliminated the antigen-specific DNA synthetic responses. The antigen-specific proliferation of murine lymphocytes depleted of adhereent cells could not be reconstituted with either guinea pig macrophages nor murine fibroblasts, indicating the existence of species and cell type specificity. In contrast to previous observations in the guinea pig, soluble products of cultured adherent cells could at least partially replace the function of intact macrophages in the response to antigen.


Safety and immunogenicity of a Neisseria meningitidis type 2 protein vaccine in animals and humans. Two Neisseria meningitidis vaccines consisting principally of outer membrane protein (lot 138I-0) or outer membrane protein plus group C polysaccharide (lot 138I-M1) were prepared from the group C type 2 strain 138I. Lipopolysaccharide and lipid were removed by gel filtration in the presence of sodium deoxycholate. The vaccines were found to be nontoxic and nonpyrogenic in animals. They provided active protection in mice against mucin-enhanced killing by group B type 2 meningococci and induced good titers of type-specific bactericidal and hemagglutinating antibodies in rabbits. In five volunteers the vaccines were well tolerated and induced significant increases in serum bactericidal activity against both group C and group B strains. Three of five volunteers had a two- to fourfold increase in antibodies to the outer membrane proteins, but these antibodies did not appear to have bactericidal activity. The bactericidal antibodies to both group B and group C strains were directed against the capsular polysaccharides.


Conjugation of coxsackievirus type B1-B6 immunoglobulins with fluorescein isothiocyanate by a "reversed" dialysis method. Immunoglobulins from antisera to coxsackievirus B types 1--6 were labeled with fluorescein isothiocyanate by a "reverse" dialysis method of conjugation. Conjugates thus obtained were labeled at reproducible ratios of fluorescein to protein with weight-weight ratios ranging from 5.0 to 7.1 and estimated molar ratios ranging from 2.1 to 2.9. Conjugates were tested on cover-slip HeLa cell cultures by the direct method of staining. Two types of specific immunofluorescence were observed: intensely fluorescent perinuclear masses and discrete foci of bright fluorescence scattered throughout the cytoplasm. Titers of homotypic conjugates varied from 1:40 to 1:160. Heterotypic staining, consisting of a diffused hazy fluorescence, was observed only at the highest concentrations of conjugate and not above the 1:10 dilution. This technique proved to be specific and sensitive for the detection of group B coxsackievirus antigens in HeLa cells but failed in specific detection of the same antigens in samples of the myocardium obtained by biopsy of three patients clinically ill with idiopathic myocarditis.


Secular variation in frequency of premature ventricular contractions in untreated individuals. Six-hour electrocardiographic monitorings were initially recorded from 40 ambulatory adult subjects (39 male, one female) on successive days. Thirty-eight of the subjects were monitored 6 weeks to 18 months later. Studies of paired time periods, Poisson modeling, and other statistical examinations were performed. Despite an underlying consistency in the data, there was considerable variation in PVC frequency from hour to hour, day to day, and over longer periods of time.


Concealed atrial bigeminy and trigeminy. Patterns indicative of concealed atrial extrasystoles were observed in two patients with frequent premature atrial depolarizations. In the first patient, the predominant pattern was such that most of the numbers (S) of sinus P waves between atrial extrasystoles satisfied the equation S = 3n-1, where n is any positive integer. This pattern is characteristic of concealed trigeminy. Over a sequence of 49 interectopic intervals, this patient vacillated between concealed atrial trigeminy and bigeminy. A second patient displayed a pattern characteristic of the "even number" variant of concealed bigeminy. The numbers of sinus P waves in consecutive interectopic intervals were predominantly even. These various patterns of concealed atrial extrasystoles closely resemble previously reported patterns of concealed ventricular extrasystoles.


Electrokinetic alteration of the surface of herpes simplex virus infected cells. Changes induced in the ionic composition of the surface of herpes simplex virus (HSV) infected cells were evaluated by whole cell electrophoresis and isoelectric focusing. The kinetics of the progressive decline in cell electrophoretic mobility (a measurement of negative surface charge density) after infection corresponded to the production of infection virus. The observed HSV-induced increase in cell isoelectric focusing pH demonstrated that the electrokinetic cell surface became ionically more positive. The nature of the ionic changes were investigated using a series of specific reagents in conjunction with cell electrophoresis. Infected cells had 2.4 times surface binding sites for the amino-specific reagent 4-acetamide-4'-isothiocyanostilbene-2,2'-disulphonic acid. A relationship between these virus-induced amino groups and HSV surface antigens was indicated by the similar kinetics of their appearance and electrophoretic neutralization by HSV antibodies.


Characterization of virus-like particles released from the hamster cell line CHO-K1 after treatment with 5-bromodeoxyuridine. Simple biochemical measurements have been shown to seriously overestimate the production of C-type particles by Chinese hamster ovary (CHO) cells treated with 5-bromodeoxyuridine. First, most particle-bound DNA polymerase activity released by induced cells was associated with particles which had a different density from C-type particles. Second, when labelled with radioactive uridine, induced CHO cells released small amounts of particle-bound radioactivity. Most of the radioactivity, however, was in DNA and did not sediment with the particle-bound polymerase. Thus, few particles which had RNA, an associated DNA polymerase, and the density typical of RNA tumour viruses were released by BrdUrd-induced CHO cells. In spite of this, some immature C-type forms were observed by electron microscopy in partially purified preparations of DNA polymerase-containing particles from induced CHO cells.


Resistance relationships in Mycobacterium smegmatis ATCC 607 to phages sensitive or resistant to both chloroform and streptomycin sulphate. Four of eight mycobacteriophages did not form plaques after they were exposed to chloroform. Phages sensitive to chloroform did not produce plaques when plated on media containing 1000 microgram/ml of streptomycin sulphate. The same concentration of dihydrostreptomycin sulphate did not interfere with plaque formation. Mutants of Mycobacterium smegmatis resistant to each of the eight phages were isolated. Sensitivity or resistance to chloroform and streptomycin sulphate and phage resistant bacterial mutants may provide a basis for classifying the mycobacteriophages.


Increased plasma levels of lead in patients with amyotrophic lateral sclerosis compared with control subjects as determined by flameless atomic absorption spectrophotometry. The levels of lead in plasma were determined in 16 cases of amyotrophic lateral sclerosis (ALS) and 18 control subjects, using flameless atomic absorption spectrophotometry. The mean values were 0.52+/-0.22 microgram/100ml (ALS) and 0.37+/-0.13 microgram/100ml (controls), the difference is statistically significant (5% level). The values in both groups are lower than reported earlier for normal subjects. The findings are discussed against the background of the possible pathogenetic significance of retrograde axoplasmic flow in ALS.


Uptake and retrograde axonal transport of horseradish peroxidase in regenerating facial motor neurons of the mouse. Uptake and retrograde axonal transport of intravenously injected horseradish peroxidase (HRP) was studied during regeneration after a crush injury of the facial nerve of the mouse. The circulation time of HRP was 12 to 24 h. HRP injected immediately after the crush diffused into injured axons in the crushed region and accumulated subsequently in perikarya of facial neurons in the brain stem. After a time interval of 1 h or 5 days between the crush and the injection only a faint HRP accumulation occurred in a few facial neurons. After an interval of 7 days a moderate number of neurons had incorporated the tracer, while after more than 9 days the HRP activity in the regenerating neurons was more pronounced than in the contralateral neurons. Ultrastructurally, muscles of the vibrissae showed denervated subneural apparatuses 6 days after the crush. 8 days after the crush regenerating axon terminals containing small clusters of synaptic vesicles, dense cored vesicles and some HRP-labelled vesicles, were found over some gutters and after 10 to 13 days all examined gutters contained axon terminals with large numbers of synaptic vesicles and some HRP-containing vesicles. More than one axon terminal profile was seen in the same synaptic gutter. 32 and 64 days after the crush the neuromuscular junctions had regained a more mature appearance. The calibre spectra of the crushed facial nerves still showed a shift towards smaller diameters 134 days after the crush, at a time when a slight increase in HRP activity in the facial neurons persisted.


Radiotherapy combined with procarbazine, bleomycin, and CCNU in the treatment of high-grade supratentorial astrocytomas. Twenty consecutive patients with supratentorial Grade III and IV astrocytomas were treated with postoperative radiotherapy, and a simultaneous induction chemotherapy with 1-(2-chloroethyl)-3-cyclohexyl-1-nitrosourea (CCNU), procarbazine, bleomycin, and maintenance chemotherapy with CCNU. They were compared to 32 retrospective control patients selected similarly, who had received postoperative radiation therapy alone. The median survival time was 56 weeks for the combined treatment group, and 51 weeks for the control group. There was no statistical difference in survival in the two groups of patients.


Quantitative determinations of two decongestants and an antihistamine in combination using paired ion high-pressure liquid chromatography. A single method for the quantitative determinations of three active ingredients, phenylephrine hydrochloride, phenylpropanolamine hydrochloride, and brompheniramine maleate, and one inactive ingredient (sodium benzoate) in a commercial product for colds is reported. The method is based on paired ion high-pressure liquid chromatography using 1-heptanesulfonic acid as the counterion. It is accurate and precise. The relative standard deviations based on six readings are reported. This method is sensitive; less than 1 microgram of each ingredient can be assayed. The peak area of each ingredient is related to its concentration.


Inhibitors of neuronal monoamine uptake. 2. Selective inhibition of 5-hydroxytryptamine uptake by alpha-amino acid esters of phenethyl alcohols. A series of alpha-amino acid esters of substituted phenethyl alcohols was prepared and tested as inhibitors of the neuronal reuptake of noradrenaline and 5-hydroxytryptamine. Some of the compounds are potent and very selective in blocking the 5-hydroxytryptamine uptake, as evidenced by biochemical data and behavioral tests. The most promising agent, alaproclate [2-(4-chlorophenyl)-1,1-dimethylethyl 2-aminopropanoate hydrochloride (I, IV)], was selected for further studies as a potential antidepressant agent. A discussion on structure--activity relationships (SAR) is given. In an attempt to explain the selective action on the mechanism of 5-hydroxytryptamine uptake by the new inhibitors, their structures are compared with those of the two neurotransmitters. From the tentative pharmacophore and conformations of transmitter (5-HT) and inhibitor (alaproclate) derived from SAR, a hypothetic carrier site for 5-HT uptake is deduced in terms of geometry and electronic properties.


An audiovisual program in cell biology.

An audiovisual program in cell biology. Cell biology has been divided into 19 topics for the purpose of planning audiovisual materials. One of these topics, the structure and function of cell membranes, has been developed as a series of seven self-instructional slide-tape units and tested in five medical schools. Organization of advisers, analysis and definition of objectives and content, and development and evaluation of scripts and storyboards are discussed.


Ionic selectivity, saturation, and block in gramicidin A channels. II. Saturation behavior of single channel conductances and evidence for the existence of multiple binding sites in the channel. A theory, recently developed by Sandblom, Eisenman and Neher (1977) for the conductance of single gramicidin A cha-nelspred icts three limiting behaviors of the relation between conductance and salt concentration. These are: (i) a saturating behavior resembling a simple adsorption isotherm at medium and high concentrations, (ii) a decrease in conductance at the highest obtainable concentrations and (iii) deviations from the isotherm at very low concentrations. Features i and ii have been described before. Experimental evidence for point iii is given here. The new feature points towards interactions among ions in the channel at ionic concentrations as low as 1--10 mM. Particular emphasis is given to the behavior at very low salt concentrations and the experimental problems encountered in this situation. In addition, mutual blocking effects among monovalent ions in symmetrical salt mixtures are characterized and found to be in satisfactory agreement with theoretical expectations, based upon the single salt conductance data presented here and zero-current potentials in salt mixtures to be described in a subsequent paper.


Malar/orbital/zygomatic fracture causing fracture of underlying coronoid process. In 1 week two patients with depressed fractures of the malar/orbital/zygomatic complex presented with roentgen evidence of fractures of the underlying coronoid process caused by the transmitted impact of the malar/orbital/zygomatic fragment without direct injury to the mandible. No similar cases had been seen in the previous 12 years, and reports found in the literature stated that this type of fracture occurs in only about 2% of all mandible fractures. Conservative treatment is recommended unless there is obvious displacement.


Binding of tRNA to reverse transcriptase of RNA tumor viruses. The interaction of tRNA with the reverse transcriptase (RNA-dependent DNA polymerase) of mammalian RNA viruses, such as Moloney murine leukemia virus and simian sarcoma virus, has been studied. Whereas the purified reverse transcriptase of mammalian viruses sedimented in glycerol gradients as a globular protein with a molecular weight of 70,000, after interaction with tRNA the enzyme cosedimented with a protein of 150,000 molecular weight. The twofold increase in molecular weight could be a result of either two reverse transcriptase molecules complexed with a tRNA or, alternatively, several tRNA molecules bound to a single enzyme polypeptide. The enzyme complexes were dissociated in part upon degradation of the tRNA moiety by pancreatic RNase A. The reverse transcriptase released from virions of Moloney murine leukemia virus, simian sarcoma virus, and avian myeloblastosis virus, by nonionic detergent, migrated faster on glycerol gradients than purified enzyme preparation. This phenomenon was probably due to complex formation between part of the virion enzyme and the tRNA, which is endogenous in virions. Addition of exogenous tRNA was needed, however, to quantitatively complex all the virion reverse transcriptase of Moloney murine leukemia virus and simian sarcoma viruses. The reverse transcriptase of Moloney murine leukemia virus did not show tRNA species specificity in the binding reaction when glycerol gradients were used for assay. Thus, several tRNA species of Escherichia coli, yeast, chicken, and rat origin were able to complex with the enzyme. The species specificity in the interaction between tRNA and avian myeloblastosis virus reverse transcriptase was also examined. We demonstrated that under our experimental conditions, this enzyme binds different tRNA species of E. coli and yeast as well as tRNA of chicken origin.


Abrogation of Fv-1b restriction with murine leukemia viruses inactivated by heat or by gamma irradiation. Fv-1b restriction in BALB/3T3 cells is temporarily abrogated following infection with N-tropic murine leukemia virus. The mechanism of this phenomenon was investigated by comparing the inactivation rates for viral infectivity and for the ability of the same virus to abrogate Fv-1 restriction. Inactivation of the abrogating ability of N-tropic murine leukemia virus following graduated doses of gamma radiation proceeded at half the rate of that for viral infectivity. This result indicates that viral RNA must function in abrogating Fv-1b restriction but that only a portion of the viral genome is required. The inactivation kinetics of N-tropic murine leukemia virus were also determined following incubation of virus at 43 degrees C. Abrogating ability of N-tropic murine leukemia virus was found to be about six times as stable under these conditions as was viral infectivity. Interestingly, virion-associated reverse transcriptase activity was inactivated at the same rate as was viral infectivity, indicating that this enzyme may not need to function during abrogation. Virus heated at 43 degrees C was used to study the kinetics of the abrogation phenomenon itself. Abrogation was shown to be transient, requiring 6 to 9 h after virus infection to become maximally effective and beginning to disappear after about 18 h. The data reported here confirm the idea that abrogation of Fv-1 restriction can be separated experimentally from virus replication, and they raise the possibility that a separate biochemical pathway exists for incoming viral RNA in Fv-1 restrictive cells.


Primate retroviruses: envelope glycoproteins of endogenous type C and type D viruses possess common interspecies antigenic determinants. The major 70,000- to 80,000-molecular-weight envelope glycoproteins of the squirrel monkey retrovirus, Mason-Pfizer monkey virus, and M7 baboon virus and the related endogenous feline virus, RD114, were isolated and immunologically characterized. Immunoprecipitation and competition immunoassay analysis revealed these viral envelope glycoproteins to possess several distinct classes of immunological determinants. These include species-specific determinants, group-specific antigenic determinants unique to endogenous primate type C viruses, and group-specific determinants for type D viruses such as Mason-Pfizer monkey virus and squirrel monkey retrovirus. In addition, a class of broadly reactive antigenic determinants shared by envelope glycoproteins of both type C viruses of the baboon/RD114 group and type D viruses of the Mason-Pfizer monkey virus/squirrel monkey virus group are described. Other mammalian oncornaviruses tested, including isolates of nonprimate origin and representative type B viruses, lacked these determinants. The demonstration of antigenic determinants specific to envelope glycoproteins of type C and type D primate viruses indicates either that these viruses are evolutionarily related or that genetic recombination occurred between their progenitors. Alternatively, endogenous type D oncornaviruses may be replication defective, and acquisition of endogenous type C viral genetic sequences coding for envelope glycoprotein determinants may be necessary for their isolation as infectious virus.


Proteins of Rous-associated virus 61, an avian retrovirus: common precursor for glycoproteins gp85 and gp35 and use of pactamycin to map translational order of proteins in the gag, pol, and env genes. Cells infected by Rous-associated virus 61 (RAV-61) contained a precursor-like protein, pr90, that was specifically precipitated by antiserum directed against envelope glycoproteins, gp85 and gp35. Tryptic peptide mapping showed that pr90 contained tryptic sequences of both gp85 and gp35. Pactamycin mapping experiments indicated that the two glycoproteins are translated from the env-mRNA in the order (5') gp85--gp35. The pactamycin mapping experiments also indicated a translational order of p10--(p27, p12)--p15 for the gag proteins; this agreement with the order previously reported from tryptic mapping studies on precursor pr76 of avian myeloblastosis virus implied that the stoichiometry of the core proteins was unchanged when virions were assembled in the presence of pactamycin. The reverse transcriptase proteins, unlike those of the env and gag genes, fell on the right side of the pactamycin map. This result is in accord with the idea that most, if not all, of the reverse transcriptase protein is translated by read-through of the gag(pol) message rather than by translation of a hypothetical pol-mRNA devoted solely to synthesis of that protein.


Unwinding-like activity associated with avian retrovirus RNA-directed DNA polymerase. The avian retrovirus RNA-directed DNA polymerase contains an activity that is capable of removing hydrogen bonds from duplex nucleic acid molecules. This "unwinding-like" activity appears to be specific in its action, affecting RNA.DNA and DNA.DNA duplex molecules but not RNA.RNA duplexes. Studies with defined RNA.DNA hybrid molecules (e.g., Rous sarcoma virus RNA and complementary DNAs representing specific regions of the Rous sarcoma virus genome) and DNA.DNA duplexes indicate that, although this activity can remove a portion of the hydrogen bonds from these double-stranded structures, complete separation of complementary strands is not accomplished. The unwinding-like activity exhibits sensitivities to temperature and monovalent and divalent cation concentrations. It can also remove a specific large oligonucleotide from the 5' end of the viral genome subsequent to RNase H hydrolysis of viral RNA complexed to DNA present at that terminus. This reverse transcriptase-associated unwinding-like activity is discussed with respect to recently proposed models of retrovirus proviral DNA synthesis.


Interstrain variation of the major internal structural component (p30gag) of two murine oncornaviruses: comparative immunochemical, biochemical, and biophysical analysis. The major internal structural protein (p30(gag)) of the Moloney leukemia virus and the endogenous Y-1 murine oncornavirus was examined for biochemical and biophysical manifestations of interstrain antigenic variation. Although the two viral proteins share murine group-specific antigenic determinants, the Y-1 virus p30 appeared to have both a lower relative number of such determinants and a decreased affinity at the cross-reactive sites for Moloney virus p30 monospecific antibodies. Further, immunological analysis indicated the presence of unique antigenic sites on the Moloney virus p30 not shared by the analogous Y-1 virus molecule. The two polypeptides copurified and had similar isoelectric points (pH 6.2 to 6.3) and sedimentation coefficients (2.47S). However, equilibrium sedimentation yielded a significant mass difference between the two proteins, 28,300 +/- 600 and 31,000 +/- 300 daltons for the Moloney and Y-1 virus molecules, respectively. Amino acid analysis indicated a concomitant increase in total residues for the Y-1 virus p30, although a number of residues appeared to have been conserved between the two viral proteins. Conformational studies and hydrodynamic calculations demonstrated marked secondary and tertiary structural differences; with the Y-1 virus p30 being an asymmetric prolate ellipsoid containing 27 to 28% alpha-helix and Moloney virus p30 being somewhat more spherical and possessing an alpha-helical content of 50 to 55%. Two-dimensional mapping of (125)I-labeled tryptic peptides of each p30 suggested that considerable sequence heterogeneity is responsible for many of the biophysical, biochemical, and immunochemical differences in these two analogous structural proteins.


Production of "rapid-harvest" Moloney murine leukemia virus by continuous cell culture on synthetic capillaries. Moloney murine leukemia virus was harvested automatically within 60 min of release from chronically infected NIH/3T3 cells (clone 1) cultured on bundles of synthetic capillaries. Production of virus as measured by a determination of reverse transcriptase activity and by the XC syncytia assay demonstrated that highly infectious "rapid-harvest" virus was recovered from NIH/3T3 cells (clone 1) grown for periods of up to 10 days.


Altered androgen metabolism in metastatic prostate cancer. Admixture of androgen-sensitive elements from normal or hyperplastic prostatic tissue interferes with biochemical studies of prostate cancer in its primary site. Heterogeneity of cancer tissues, varying in stromal and epithelial elements, also complicates interpretation of data relating to androgen metabolism. Accordingly, we have compared metastatic deposits composed of epithelial cancer cells to the primary biopsies of 4 patients in respect to uptake of 3H-testosterone and its conversion to 5-alpha-dihydrotestosterone during in vitro incubation. 3H-testosterone uptake was similar for both tissue sites but 3H-dihydrotestosterone formation was reduced by 76% in the metastases compared to primary tissues. This group was not large enough to show statistical significance, whereas a total of 11 such primary studies compared to 6 metastatic specimens was significant. When either primary or secondary tissue results were compared to 12 cases of benign prostatic hyperplasia similarly studied the differences were highly significant. These results demonstrate a major impairment in the formation of dihydrotestosterone by metastatic prostatic cancer and a similar but less evident alteration in the primary site. This abnormality in testosterone metabolism is of major importance in the attempt to obtain effective hormonal control of human prostatic cancer.


Direct recording of urethral resistance using the "urethroresistance". Observations on urethral resistance during micturition in non-obstructed men and in patients with bladder outlet obstruction owing to benign prostatic hypertrophy are presented. The "Urethroresistance", a new instrument, was used for the direct recording of urethral resistance. The upper limit for the minimum urethral resistance during micturition for normal non-obstructed men was 0.5 units. Two patterns of urethral resistance were observed solely in non-obstructed subjects and 3 other patterns, which were found solely in patients with a moderate or severe degree of bladder outlet obstruction, were shown to be characteristic of bladder outlet obstruction. While direct recording of urethral resistance is a better test for bladder outlet obstruction than the recording of urinary flow rate the recording of urinary flow is a simple urodynamic test, which remains a useful screening test for bladder outlet obstruction. With the exception of severe obstructive symptoms none of the other clinical methods for the assessment of bladder outlet obstruction--the size of the prostate gland, the volume of post-micturition residual urine and the degree of bladder trabeculation--is reliable to determine the degree of bladder outlet obstruction.


Self-retained internal ureteral stents: a new approach. Definitive surgical treatment of ureteral obstruction may not be appropriate in patients with advanced malignancy, complex benign ureteral disease or even simple obstructive disease accompanied by unacceptable operative risk. Endoscopic placement of self-retained internal ureteral catheters (stents) offers satisfactory relief for many of these patients. A new 6F pigtail ureteral stent and placement technique are described. These stents are placed easily, effective and well tolerated for short-term drainage in selected patients with ureteral obstruction.


The role of the radioimmunoassay of serum alpha-fetoprotein and human chorionic gonadotropin in the intensive chemotherapy and surgery of metastatic testicular tumors. Quantitative measurement of serum alpha-fetoprotein and human chorionic gonadotropin by double antibody radioimmunoassays reflects the efficacy of surgical, radiation and/or chemotherapeutic regimens in patients with bulky disseminated testicular tumors. When these therapies are effective they produce an immediate decrease in serum levels of these markers that reflects the decrease in tumor size and could be as rapid as the catabolic decay rate for alpha-fetoprotein or human chorionic gonadotropin. The alpha subunit of human chorionic gonadotropin has a short half-life (20 minutes) and has been used for the first time to localize a recurrent metastatic testicular tumor. Cellular localization of these markers by immunochemical techniques has helped us to understand the natural history of this cancer and the cell types responsible for production of the markers.


The effect of human serum on 3H-thymidine incorporation in human prostate tumors in tissue culture. Human prostatic tumors, 4 adenomas and 4 adenocarcinomas, were established in tissue culture. We used the incorporation of 3H-thymidine into deoxyribonucleic acid as a measure of cell activity to compare the effects of supplementing the culture media with human or fetal calf serum. 3H-thymidine incorporation was greater with human serum, with up to 120 per cent increase observed. The addition of dihydrotestosterone (10(-8)M.) did not enhance or inhibit 3H-thymidine incorporation.


Pharyngeal gram stains in the treatment of sore throats. The use of the direct pharyngeal Gram stain for diagnosis of group A beta-hemolytic streptococcal pharyngitis was evaluated. Seventy-five consecutive out-patients presented with pharyngitis (or high fever in infants) were studied with pharyngeal cultures and Gram stains. Gram stains were read as either positive, negative, or indeterminate for streptococcal pharyngitis. When these predictions were correlated with the resulting culture growth of group A beta-hemolytic streptococci, the predictive value of a throat smear Gram stain was found to be significant (p less than 0.005). There were two false positives and three false negatives. Approximately one third of the smears were in the indeterminate category. Although the Gram stain of a pharyngeal smear was found to be less sensitive than the routine throat culture, the ability to obtain inexpensive rapid microbiological information suggests that the Gram stain can be used as a quick screening procedure.


Blood-pressure and mortality in a rural Jamaican community. 1065 men and women aged 35--64 years living in rural Jamaica were first examined in 1962--1963, re-examined 5 and 10 years later, and followed-up until 1976. Overall mortality in 13 years, taking into account blood-pressures at all 3 surveys, showed that mortality was significantly increased only at pressures above 180 mm Hg systolic or 110 mm Hg diastolic. It was estimated that without this degree of hypertension the total number of deaths between the ages of 45 and 69 years would have been reduced by about 17%. Unlike other reports, mortality showed no significant association with lower levels of blood-pressure; this difference may be due to a lower incidence in this community of coronary and cerebral thrombosis. The relation between blood-pressure and mortality cannot be assumed to be identical in populations with different profiles of cardiovascular pathology.


Importance of pituitary hormones in aetiology of diabetic ketoacidosis. The role of pituitary hormones in the aetiology of diabetic ketoacidosis was examined by withdrawing insulin from five pituitary-ablated diabetics for a 12-hour period. The rise in blood glucose and ketone-body concentrations was markedly retarded in these patients when compared with five matched juvenile-type diabetic controls with normal pituitary function. When cortisol replacement in the pituitary-ablated patients was increased to reproduce the high plasma concentrations found in severe ketoacidosis, blood ketones and glucose were increased but were still significantly lower than in the control diabetics. It is concluded that pituitary hormones may be important in the development of diabetic ketoacidosis.


Circulating lymphocytes and the spread of myeloma. Review of the evidence. Patients with myeloma harbour circulating lymphocytes which bear the idiotypic determinants (SIg-id+ cells) of their own myeloma protein. Circulating white cells with abnormal karyotypes have been found in other patients with this disease. SIg-id+ lymphocytes have also been identified in the peripheral blood of myeloma-tumour-bearing mice. Murine myelomas have been propagated in vivo from circulating mononuclear cells that possessed myeloma-tumour-associated antigens. This and other evidence reviewed here points strongly to the involvement of circulating lymphocyte-like stem cells in the spread of human myeloma.


Asymptomatic endemic rotavirus infections in the newborn. Between May 1, 1976, and May 14, 1977, 343 (32.5%) of 1056 5-day-old babies in newborn nurseries excreted rotaviruses. The infection-rate was highest during winter (49%). 76% of infected babies at this time were bottle-fed. 41% of neonates excreted low amounts of virus (less than or equal to 10(8) particles/g faeces); older children tended to excrete greater than 10(10) particles/g faeces. Infected breast-fed babies excreted less virus than those who were bottle-fed. Stools of breast-fed babies often contained clumps of complete "smooth" rotavirus particles. When the newborn nurseries were transferred to a newly built hospital wing, infection appeared in the new wards, including those admitting only new patients, within a short period. Infection was either mild (8%) or symptomless (92%), and even babies with symptoms required no treatment.


Prevention of iron loading in transfusion-dependent thalassaemia. Urinary iron excretion after single intramuscular (i.m.) bolus injections or 12 h subcutaneous (s.c.) infusions of desferrioxamine (D.F.) was determined in sixteen homozygous beta-thalassaemia patients whose ages ranged from 10 months to 23 years. At all ages the s.c. infusions resulted in greater iron loss than identical i.m. doses. With doses of 0.5-1 g of D.F. as s.c. infusions eight out of nine children aged less than 6 years with a total transfusion iron load of less than 10 g excreted sufficient iron to achieve iron balance. These results suggest that iron loading in transfusion-dependent thalassaemics may be preventable by the use of an appropriate chelation regimen started early in life.


Colonic function in patients with diverticular disease. Diverticular disease is thought to be associated with prolonged intestinal transit-time, a reduced stool weight, and increased intracolonic pressure. Sixty patients with diverticular disease did not regularly show these features. Variation in colonic function was considerable in these patients and was similar to that in the general population from which the patients were recruited. Constipation may be a complication of diverticular disease and not necessarily part of its aetiology.


Extensive prolongation of rabbit kidney allograft survival after short-term cyclosporin-A treatment. The fungal metabolite cyclosporin A has been administered daily at 25 mg/kg to nephrectomised rabbit recipients of a single renal allograft. After only 4 weeks of daily administration, rejection was prevented for long periods without the need for other immunosuppressive therapy. Renal function has been excellent in all cases, rejection episodes have not been encountered except in a small percentage of animals, and the surviving recipients are still in excellent health 3-6 months after transplantation. The possiblity that clones of responding lymphocytes might be selectively killed by this agent was suggested from earlier experiments and prompted this study. If toxicity is not encountered in man cyclosporin A may prove valuable in preventing rejection of organ and bone-marrow allografts and might be useful in treating some leukaemias of lymphoid origin.


Prolonged survival of pig orthotopic heart grafts treated with cyclosporin A. Cyclosporin A was given to pigs receiving orthotopic cardiac allografts from donors mismatched at the major locus. Median survival in 20 control pigs was only 6 days; in 5 pigs given cyclosporin A at 15 mg/kg on 3 days median survival was 22 days; and when 6 animals were given 25 mg/kg intramuscularly for 2 days and then orally median survival is greater than 68 days, with 4 animals still alive.


Scintigraphic detection of atherosclerotic lesions and venous thrombi in man by indium-111-labelled autologous platelets. Accumulation of autologous platelets labelled with indium-111 at sites of atherosclerosis or venous thrombosis was demonstrated by scintigraphy in three patients. The lesions detected were bilateral ulcerated carotid-artery plaques, iliofemoral venous thrombosis, and renal-vein thrombosis. This technique shows promose as a non-invasive means of diagnosing focal atherosclerotic and thrombotic lesions.


Prospective study of the treatment of septic shock. A study of 113 patients with septic shock admitted to an intensive therapy unit (I.T.U.) during a 3-year period showed that a significant reduction in mortality was achieved after the introduction of a treatment programme which included early intermittent positive-pressure ventilation and aggressive surgery in addition to conventional management with fluids, oxygen, and antibiotics. Vasoactive drugs played an important but not essential role in therapy.


Reduced high-density lipoprotein in women aged 40-41 using oral contraceptives. Consultation Bureau Heart Project. In a survey of risk factors for coronary heart-disease (C.H.D.) in a Leiden population aged 40-41 years, mean serum-high-density-lipoprotein (H.D.L.)-cholesterol was significantly higher in 447 women (47.94 mg/dl) than in 471 men (42.40 mg/dl). No association was found between serum-H.D.L.-cholesterol and systolic or diastolic blood-pressure, obesity, or electrocardiographic changes. Cigarette smoking with use of oral contraceptives were strongly associated with reduced serum-H.D.L.-cholesterol. The difference in mean serum-H.D.L.-cholesterol concentrations between women who were on oral contraceptives and those who were not was independent of the effect of smoking. The finding of a low mean serum-H.D.L.-cholesterol concentration in pill users who smoke (i.e., similar to that in men of the same age; 43.0 mg/dl and 42.4 mg/dl in women and men, respectively) is disturbing since low serum-H.D.L.-cholesterol is a major C.H.D. risk factor and because of the reported increase in mortality from circulatory diseases in women using oral contraceptives.


Very early abortion by prostaglandins.

Very early abortion by prostaglandins. 309 women whose menstruation was delayed by 3-35 days were treated with intrauterine or vaginal prostaglandins. Of 275 confirmed pregnancies, 229 were successfully terminated without further abortifacient therapy. A successful outcome was often associated with episodes of vomiting, diarrhoea, and uterine cramps in the 24 hours after prostaglandin administration, but the incidence was related to prostaglandin dosage and gastrointestinal side-effects were more common after vaginal administration. The best results were achieved by the analogue 16:16 dimethyl P.G.E2 as a vaginal pessary. 14 patients (6.1%) required uterine curettage for escessive or prolonged bleeding, while 2 patients required blood transfusion. One patient, who had an intrauterine contraceptive device left in situ during treatment, developed acute pelvic sepsis. No deleterious side-effects occurred in 34 patients who were subsequently proven not to be pregnant at the time of treatment. Treatment by intrauterine or vaginal prostaglandins offers promise as a method of pregnancy termination which avoids much of the physical and emotional trauma associated with surgical termination, and has the advantage of not requiring hospital admission in the majority of cases. The present study shows the safety of the method, and its potential as a self-administration technique.


Zinc and immunocompetence in protein-energy malnutrition. Malnourished children have thymic atrophy which is reversed by zinc supplementation. To see if their defect in cell-mediated immunity was also associated with zinc deficiency ten children were skin-tested with Candida antigen on both arms. One test site was covered with local zinc sulphate and the other with placebo ointment. There was a highly significant increase in the typical delayed-hypersensitivity reaction at the site covered with zinc. The magnitude of the difference between the supplemented and unsupplemented arms correlated negatively with the plasma-zinc concentration. These data show that zinc deficiency is a cause of the immunoincompetence seen in malnutrition. The normal reactions of the zinc-supplemented side indicate that, of the many nutritional deficits of malnourished children, zinc deficiency specifically impairs the cell-mediated immune system. Local skin-testing with and without zinc may provide a measure of zinc status. Local application of zinc may enhance the reliability of tests to diagnose diseases such as tuberculosis in malnourished patients.


Enhancement of human kidney allografts by cold B-lymphocyte cytotoxins. The sera of 233 kidney transplant patients before transplantation were tested by cytotoxicity against a panel of B and T lymphocytes at 5 degrees C and 37 degrees C. The results divided the patients into four groups: those whose sera reacted with B lymphocytes at 5 degrees C; those reacting with B lymphocytes at 5 degrees C and 37 degrees C; those reacting with T lymphocytes at 37 degrees C; and those with no antibodies. The patients with pre-transplant antibodies reactive with B lymphocytes at 5 degrees C had a significantly higher kidney-transplant survival rate at 6 months (70%) and 1 year (65%) than patients who had no antibodies (47% and 46%, respectively). Patients with antibodies reactive at 37 degrees C had a 6-month survival-rate of 38% when reactive against B cells and 43% when reactive against T lymphocytes. The cold cytotoxins were IgM.


Increased adipose-tissue lipoprotein-lipase activity in moderately obese men after weight reduction. Most of the many metabolic abnormalities associated with obesity are corrected by weight reduction. Adipose-tissue lipoprotein-lipase activity per cell is increased in obesity. Adipose-tissue lipoprotein-lipase was significantly higher in seven previously obese men studied at a stable reduced weight than in a control population. In fact, enzyme activity was significantly higher than would have been predicted from the obese men's maximum weight. These results indicate that abnormalities in adipose-tissue lipoprotein lipase may have a primary role in the development of obesity.


Sucrose malabsorption in man after ingestion of alpha-glucosidehydrolase inhibitor. The effect of an alpha-glucosidehydrolase inhibitor (alpha-G.H.I.) on intestinal absorption of sucrose was assessed by measuring rises in blood-glucose and intestinal hydrogen production after ingestion of 100 g sucrose. 200 mg alpha-G.H.I. given with the sucrose completely inhibited early postprandial blood-glucose rises, prevented late postprandial dips in blood-glucose, and considerably increased breath-hydrogen levels, which indicates sucrose malabsorption. Thus the inhibitor can be used to study carbohydrate malabsorption and may be useful as a method for retarding carbohydrate absorption.


Vasopressin in affective illness.

Vasopressin in affective illness. Animal studies have revealed two important aspects of vasopressin function which make this peptide a suitable candidate for involvement in complex behavioural syndromes: (1) vasopressin deficiency produces deficits of behaviour which are reversed by vasopressin; (2) well-developed systems exist for the distribution of vasopressin throughout the central nervous system (C.N.S.) via either peptidergic neurons or the cerebrospinal fluid (C.S.F.) and provide the means by which vasopressin may regulate cells controlling behavioural or physiological processes. Among the processes which vasopressin can influence are several of significance in the symptom-complex of affective illness, including alterations in memory, changes in pain sensitivity, synchronisation of biological rhythms, the timing and quality of R.E.M. sleep, and the regulation of fluid and electrolyte balance. In addition, vasopressin is functionally linked to monoamine neurotransmitter systems and, like them, is altered by pharmacological agents which affect mood. Some of the pharmacological and clinical data suggest that vasopressin function is diminished in depression and augmented in mania; sometimes, however, alterations in vasopressin function may be detectable only during crucial periods of the manic-depressive cycle. The hypothesis that vasopressin plays a role in disorders of human behaviour, particularly manic-depressive illness, can now be directly tested by radioimmunoassays of vasopressin in C.S.F. and plasma and by the administration of specific vasopressin analogues and inhibitors.