A soluble acidic protein of the cell nucleus which reacts with serum from patients with systemic lupus erythermatosus and Sjögren's syndrome. A soluble nuclear antigen that reacts with sera obtained from patients with systemic lupus erythematosus and Sjögren's syndrome has been described. The antigen, tentatively named the Ha antigen after the prototype serum, was shown to react with specific antibodies by precipitin, complement fixation, and immunofluorescence techniques. The Ha antigen prepared from isolated nuclei of calf thymus glands, calf liver, and rat liver showed identical immunological reactivities; a wide distribution among different species and tissues is presumed. The Ha antigen was destroyed by trypsin and relatively mild heat or pH variation from neutrality, but was resistant to DNase or RNase. Many of these characteristics are similar to those of the "B" antigen to which antibodies have recently been described in Sjögren's syndrome. The  nuclear origin of the Ha antigen was confirmed by the speckled nuclear immunofluorescence staining pattern given by purified antibody to Ha obtained from a specific immune precipitate. Preliminary results showed approximately 13% of patients with systemic lupus erythematosus and 30% of patients with Sjögren's syndrome had precipitating antibodies to the Ha antigen.


The Minnesota Child Development Inventory ad an aid in the assessment of developmental disability. Parents' reports on their children's functioning were obtained from the Minnesota Child Development Inventory (MCDI) during a review of 132 cases. These data were found to be highly correlated (r = .92) with mental ages obtained during formal psychometric evaluation. The data support the use of the MCDI as an additional tool in the assessment of clinical population of children suspected of having developmental disabilities.


Purified azure B as a reticulocyte stain. A comparison has been made between reticulocyte preparations stained with purified azure B and with several commerically available batches of brilliant cresyl blue and new methylene blue. Marked variations were observed in the composition and staining performances of the various batches of the two commerically available dyes. Although there were no significant differences in reticulocyte counts obtained with these two dyes, varying amounts of an extraneous, particulate dye deposit were present in these preparations, making accuracte counting both tedious and timeconsuming. Purified azure B, on the other hand, gave reproducibly stained, deposit-free preparations. Reticulocyte counts obtained from azure B preparations  correlated almost exactly with those determined using new methylene blue. Purified azure B is therefore recommended as a convenient reticulocyte stain for routine use.


Anatomical studies of a temporal visual area in the rabbit. Using Fink-Heimer, autoradiographic and horseradish peroxidase techniques, the connections of a temporal visual cortical region of the rabbit were explored. The temporal visual area covers portions of areas T1 and T2, and is reciprocally connected with the posterior nucleus and suprageniculate nuclei of the thalamus. It was also shown that the temporal visual area projects to a similar region in the opposite hemisphere, and to intermediate laminae of the superior colliculus. The temporal visual area is discussed in comparison to other similar regions in the cortex of primate species. It is pointed out that recent evidence indicates visual areas in the occipital cortex of non-primate species may be no less numerous and complex than those in primate species.


The cortical projections of the mediodorsal nucleus and adjacent thalamic nuclei in the rat. The mediodorsal nucleus of the rat thalamus has been divided into medial, central and lateral segments on the basis of its structure and axonal connections, and these segments have been shown by experiments using the autoradiographic method of demonstrating axonal connections to project to seven distinct cortical areas covering most of the frontal pole of the hemisphere. The position and cytoarchitectonic characteristics of these areas are described. The medial segment of the nucleus projects to the prelimbic area (32) on the medial surface of the hemisphere, and to the dorsal agranular insular area, dorsal to the rhinal sulcus on the lateral surface. The lateral segment projects to the anterior cingulate area (area 24) and the medial precentral area on the dorsomedial shoulder of the hemisphere, while the central segment projects to the ventral agranular insular area in the dorsal bank of the rhinal sulcus, and to a lateral part of the orbital cortex further rostrally. (The term "orbital" is used to refer to the cortex on the ventral surface of the frontal pole of the hemisphere.) A ventral part of this orbital cortex also receives fibers from the mediodorsal nucleus, possibly its lateral segment, but the medial part of the orbital cortex, and the ventrolateral orbital area in the fundus of the rhinal sulcus receive projections from the paratenial nucleus and the submedial nucleus, respectively. All of these thalamocortical projections end in layer III, and in the outer part of layer I. The basal nucleus of the ventromedial complex (the thalamic taste relay) has been shown to have a similar laminar projection (layer I and layers III/IV) to the granular insular area immediately dorsal to, but not overlapping, the mediodorsal projection field. However, the principal nucleus of the ventromedial complex appears to project to layer I, and possibly layer VI, of the entire frontal pole of the hemisphere. The anteromedial nucleus does not appear to project to layer III of the projection field of the mediodorsal nucleus, although it may project to layers I and VI, especially in the anterior cingulate and medial precentral areas. A thalamoamygdaloid projection from the medial segment of the mediodorsal nucleus to the basolateral nucleus of the amygdala has also been demonstrated, which reciprocates an amygdalothalamic projection from the basolateral nucleus to the medial segment. The habenular nuclei also appear to project to the central nucleus of the amygdala. These results are discussed in relation to the delineation and subdivision of the prefrontal cortex in the rat, and to amygdalothalamic and amygdalocortical projections which are described in a subsequent paper (Krettek and Price, '77).


A comparison of visual pathways in Boston and Midwestern Siamese cats. A genetic mutation in Siamese cats causes retinogeniculate fibers representing roughly the first 20 degrees of ipsilateral visual field in each eye to cross aberrantly in the optic chiasm and terminate in the wrong lateral geniculate nucleus (LGN). Previous investigations have shown that in the visual cortex this extra representation of ipsilateral visual field can be organized into one pattern in Boston Siamese cats, another in Midwestern. This finding was confirmed here. The possibility that the organization of the LGN might account for these two patterns was studied using combined anatomical and physiological methods. On the basis of microelectrode recordings from the visual cortex, 11 out of the 12 Siamese cats included here were Boston cats; one was Midwestern. The distribution of retinogeniculate terminals was examined in each cat using autoradiographic techniques following an eye-injection of 3H-proline. Overall, the LGN organization in Boston cats was similar to that of Midwestern: both lateral and medial normal segments of lamina A1 (mnA1) were present. In Boston cats, however, the mnA1 was remarkably small and shifted ventromedially in the nucleus to allow for the fusion between the medial borders of lamina A and the abnormal segment of A1. In the Midwestern cat this fusion was not apparent and the medial normal segment of A1 was significantly larger. These differences in organization of the LGN are consistent with those seen at the level of the visual cortex in Midwestern and Boston Siamese cats. It was not possible, however, to relate them clearly to the characteristic strabismus of these animals.


Ultrastructural studies of the superior cervical trunk of the mouse: distribution, cytochemistry and stability of fibrous elements in preganglionic fibers. The ultrastructure of axons in the preganglionic cervical sympathetic trunk of the mouse is described with emphasis on the number, distribution and stability of fibrous elements in the axoplasm. Neurofilaments outnumbered microtubules in myelinated and non-myelinated axons of all sizes, and the ratio of neurofilaments to microtubules in non-myelinated axons at each point studied was fairly consistent and independent of axonal diameter. The density of neurofilaments and microtubules, however, was greater in axons of progressively smaller diameter. In non-myelinated axons and small myelinated axons neurofilaments were uniformly distributed throughout the axoplasm resulting in minimum and maximum interfilament distances of 300 angstrom and 500 angstrom respectively; the spacing of fibrous elements within any one axon was dependent upon its diameter and position with respect to the superior cervical ganglion in the preganglionic trunk. The maximum interfilament distance was also found in large myelinated axons where neurofilaments, occurring in fascicles, were separated by distances of approximately 500 angstroms. Cytochemical staining of axons with lanthanum hydroxide, ruthenium red or alkaline bismuth delineated the delicate filamentous matrix interconnecting microtubules, neurofilaments and other organelles in the axoplasm. Alkaline bismuth stain was most intense in myelinated axons where heaviest deposition of reaction product was associated with neurofilaments. Treatment in vitro of the cervical sympathetic trunk with 5 X 10(-5) M vinblastine sulfate dissociated microtubules and induced formation of crystalline arrays of "tubular" elements. A uniform center to center spacing of 250-300 angstrom was found for crystalloids in non-myelinted axons; however, in myelinated axons the center to center spacing was not uniform and varied in the range 300-600 angstrom. Neurofilaments and their surface projections were unaffected by vinblastine. Fixation in the presence of lanthanum enhanced delineation of crystalloid elements. Exposure of 0-4 degrees C for up to three hours had no consistent effect on microtubules or neurofilaments. In contrast, cold treatment disrupted the delicate axonal matrix and resulted in the formation of aggregates of coarse flocculent material in the axoplasm.


Survival and fertility of antibiotic-treated bovine spermatozoa. Motility of spermatozoa stored at 5 C with up to 1000 units or mug of chloramphenicol, polymyxin, kanamycin, tylosin, ampicillin, lincomycin, spectinomycin, erythromycin, novabiocin, or terramycin per ml of extender was compared to that with penicillin plus dihydrostreptomycin. Novabiocin and terramycin were toxic, but other antibiotic treatments had no effect. However, erythromycin and tylosin, as well as colymycin, depressed motility of frozen thawed spermatozoa. Spermatozoal motility was equivalent, following freezing in ampules or straws. All of the antibiotics which were non-toxic when added singly to frozen semen were also not harmful to frozen spermatozoa when as much as 2000 units or mug were added per ml of extender containing penicillin and dihydrostreptomycin. The addition of 1000 units or mug of ampicillin, chloramphenicol, kanamycin, lincomycin, polymyxin, or spectinomycin per ml of extender containing 750 units penicillin and 750 mug dihydrostreptomycin per ml did not influence the fertility of frozen spermatozoa in a field test involving 19,663 first inseminations.


Harvester ant sensitivity: in vitro and in vivo studies using whole body extracts and venom. Harvester ant stings by Pogonomyrmex maricopa (Pm) or Pogonomyrmex rugosus (Pr) resulted in serious reactions in 8 patients, 4 with generalized reactions and 1 with large local reactions. Exposure to one species in the genus Pogonomyrmex (P) appeared to cross-sensitize ant-sensitive patients to other species in the same genus as evidenced by skin testing and leukocyte histamine release, but these patients were less sensitive to extracts from other stinging Hymenoptera, including bee, wasp, yellow jacket, hornet, and Formica ant. Pr ant venom was obtained by electrical stimulation of live ants for leukocyte histamine release studies. The venom preparation was considerably more effective in inducing histamine release than a body extract derived from gasters, the posterior abdominal segments. Rabbits immunized with an extract from one species produced precipitating antibodies against the injected extract withich cross-reacted with extracts from other species of harvester ant, but not with other stinging Hymenoptera. Humans and rabbits appear to react to certain genus-specific antigens present in Pogonomyrmex whole body extracts. Proper identification of the offending ant is crucial for proper testing and treatment of ant-sensitive patients.


Diagnosis of Hymenoptera hypersensitivity by skin testing with Hymenoptera venoms. This double-blind study provides evidence that skin testing with dialyzed Hymenoptera venoms is a more accurate, reliable method of diagnosing hypersensitivity to the sting of honeybee, yellow jacket, yellow hornet, white-faced hornet, and wasp than is skin testing with the corresponding whole body diagnostic allergenic extract. Furthermore, the incidence of false-positives was greatly reduced by using the dialyzed Hymenoptera venom (HDV) diagnosis. In this clinical trial, most sensitive individuals had skin test reactions greater than the diluent control at concentrations of 1 mug/ml or below. Levels of venom diagnostic of 100 mug/ml appeared to produce nonspecific local irritation. Skin test with whole body diagnostic allergenic extract did not produce a consistent differentiation between sensitive and nonsensitive individuals. No untoward reactions were seen using the dialyzed Hymenoptera venom (HDV) diagnostic.


Hypersensitivity to pancreatic extracts in parents of patients with cystic fibrosis. Because immediate hypersensitivity reactions can occur in individuals exposed to powdered pancreatic extracts, 36 patients with cystic fibrosis and 51 patents of such patients wwer studied for evidence of sensitization. Sensitivity to the extracts as evidence by history and skin testing was infrequent in the children with cystic fibrosis. However, skin testing for immediate hypersensitivity with either crude pancreatic extracts or inactivated trypsin correlated well in their patents with a history of clinical symptoms. IgE mediation of these reactions in sensitized individuals was demonstrated by antigen-induced histamine release from leukocytes, passive transfer studies, and immediate response to inhalation challenge.


Immediate hypersensitivity to cockroach. Isolation and purification of the major antigens. Crude cockroach extract elicited positive skin tests in 50% of patients with positive and in 4% with negative environmental history for cockroach exposure, suggesting a possible role of cockroach in perennial atopic disease. Three major allergens in crude American and German cockroach extracts have been identified using sequential purification steps on Sephadex G-75, diethylaminoethyl (DEAE) cellulose, and agarose gel electrophoresis. Cr-I elicits positive skin tests in 70% of patients sensitive to the crude extracts. It has a molecular weight of approximately 25,500 daltons, is highly acidic, and resists boiling for four hours. Boiling in 4 N acetic acid completely abolishes its allergenicity. The purified allergen elicits positive skin tests at a concentration of 3 mug/ml and is capable of inducing greater than 50% histamine release from sensitive leukocytes at 0.05 ng/ml. A second antigen, Cr-II, elicits positive skin tests also in approximately 70% of cockroach-sensitive individuals, has a molecular weight of approximately 63,000 to 65,000 daltons, and has similar heat stability and acid hydrolysis characteristics to Cr-I. A third, less well-characterized antigen, Cr-III, has a molecular weight less than 10,000 daltons and elicits positive skin tests in 30% of individuals sensitive to the crude extract.


Studies on Brugia pahangi. 13. The anthelmintic effect of compounds F151 (Friedheim), HOE 33258 (Hoechst) and their reaction product. F151 was a potent filaricide against adult Brugia pahangi in cats and jirds. HOE 33258 did not kill adult worms in cats but had a marginal effect on adult worms in the peritoneal cavity of jirds. It was not immediately microfilaricidal in cats but the microfilarial counts of treated cats fell within a few weeks of treatment. The reaction product, or mixture, of these two compounds (V5851 = E) was strongly macrofilaricidal in cats and jirds.


Interaction of polyanions with basic fuchsin and formaldehyde. Basic fuchsin and formaldehyde react readily with a number of polyanions in aqueous solutions. Deoxyribonucleic acid, ribonucleic acid, several synthetic polynucleotides and polyvinylsulfate all convert buffered solutions of basic fuchsin and formaldehyde from a magenta to a purple color at ambient temperature. The formation of product is maximal in about 20 min and the resulting complexes are stable enough to be isolated by gel filtration. Microgram amounts of polyanion suffice to produce the color changes and the spectral shifts vary with the type of polyanion. These results are interpreted to indicate that polymerization of basic fuchsin and formaldehyde is an essential aspect of color formation.


Simultaneous staining of ribonucleic and deoxyribonucleic acids in unfixed cells using acridine orange in a flow cytofluorometric system. Simultaneous staining of deoxyribonucleic (DNA) and ribonucleic acid (RNA) in nonfixed, but permeable, cells is described. Cells are made permeable by treatment with non-ionic detergent at low pH. RNA is denatured prior to, or during staining, by exposure of cells to chelating agents to ensure that DNA (native) and RNA (dentured) may be stained differentially with the metachromatic dye, acridine orange. The fluorescence of individual cells is measured in a flow cytofluorometer. A comparison between various staining procedures employing acridine orange or other intercalating dyes in unfixed cells is discussed in terms of staining specificity, cell permeability and preservation. Evidence is provided that acridine orange staining of unfixed cells may be used as a simple, fast means of obtaining information on cell ploidy levels and cell cycle status from DNA measurements (green fluorescence), and cell transcriptional activity from RNA staining (red fluorescence), in human and murine cells lines, peripheral blood and bone marrow specimens from patients with leukemia and mitogenically (phytohemagglutinin) or antigenically (mixed lymphocyte culture) stimulated human peripheral blood cultures. Exposure of cells to detergent at low pH as an alternative to cell fixation or hypotonic treatment is proposed as a fast, convenient method of making cells permeable to dyes.


Optical properties of normal and injured cells. Application of cytographic analysis to cell viability and volume studies. A cell spectrophotometer (Cytograf Model 6300 A, Bio/Physics Systems, Inc.) was tested in a cytotoxic assay using Ehrlich ascites tumor cells as a model system. Several cellular conditions associated with volume expansion, staining of cellular components and fixation of cells were applied and the magnitude of the scattering and extinction signals were tested in these diverse cellular conditions. The magnitude of the scattering pulse of a cell spectrophotometer was found to be greatly dependent on the staining color and intensity of cellular components with vital dyes or following osmium tetroxide fixation. When the absorption wavelength of the vital dye was close to the wavelength used in the cell spectrophotometer (about 630 nm), dead stained and living nonstained cell populations were completely separated from each other. The magnitude of the extinction pulse was greatly dependent on the state (normal, injured cells) and staining intensity (vital dye staining, osmium fixation) of cellular components. The magnitude of the extinction pulse was reduced from that in normal cells when cells were treated with p-chloromercuribenzene sulfonic acid or in a hypotonic solution that caused a marked volume expansion of injured cells. When cells were fixed with a mixture containing glutaraldehyde and osmium tetroxide the cellular components of normal and injured cells turned black and in these conditions the magnitude of the extinction signal was in a linear correlation to the cross-sectional area of cells. In the present study, the cell spectrophotometer proved to be an efficient method for estimation of cellular viability, based on different scattering properties of cells, offering the advantages of high speed and precision. Demonstration of the use of a variety of vital dyes having diverse extinction properties with capabilities to differentiate between living and dead cells has indicated the potential use of the cell spectrophotometer in cytotoxic assay. Modification of the magnitude of the extinction signal in the cell spectrophotometer also shows great promise for accurate automated size determination of both normal and injured cells. Previously, determination of the size of injured cells has been beset with methodologic errors.


A general method for the rapid separation and specific detection of antigenic meterial by immunoelectrofiltration using multispecific antisera. A general technique is described for the separation and detection of specific antigens from complex antigen mixtures, by their electrophoresis through antibody-containing gels. The method does not require purified reagent antigen or fuctionally monospecific antisera and should have a wide applicability in the detection, quantification and characterization of various antigens.


The ethanol fractionation of mouse serum. A method for the ethanol fractionation of normal mouse serum has been developed which is rapid and simple to perform. The method was shown to be effective in purification of IgG1, IgG2 and gammaM-gammaA globulins. Gel filtration and zone electrophoresis proved to be effective in separating the gammaM-gammaA globulin combination into their respective components. Immunoelectrophoresis was chosen to assess the homogeneity of the fractions and the results agree with those obtained by other workers.


Cad(super Sda) in a British family with eastern connections: a note on the specificity of the Dolichos biflorus lectin. Cad, Sd(a++) or Super Sda is a rare, inherited, dominant blood group character which is of much interest, not only with regard to problems in pretransfusion tests (erythrocyte polyagglutination) but also in the field of lectin specificity. We have studied this blood group character in a British family with Eastern connections and present a brief account of its serological and clinical importance. Most persons are Sd(a+) but there is a wide distribution of antigen strength, ranging from ordinary Sd(a+) to Cad. Most persons also have weak anti-Sda in their serum; this is ordinarily of no clinical importance, but could cause problems if Cad bloods are transfused. The chief structural determinant of Cad specificity is N-acetyl-D-galactosamine in alpha-linked position, yet it is clearly distinguishable by use of appropriate lectins from other blood group antigens, A and Tn, which also have this acetyl-hexosamine as their chief structural determinant. A method for the rapid identification of Cad, applicable to all ABO groups, is described. The lectin of Dolichos biflorus, which is specific for N-acetyl-D-galactosamine in alpha-linked position, reacts strongly with A(A1), Tn and Cad cells, its action on Cad cells being much the strongest. Absorption-elution studies show that one and the same lectin reacts with both A1 and Tn cells. Absorption with Cad cells abolishes activity for A1, Tn and Cad cells; whereas absorption with A1 or Tn cells leaves activity for Cad. This does not necessarily indicate that anti-Cad is a separate component since the same result can be obtained by simply diluting the Dolichos reagent. However, eluates from Cad cells react only with Cad cells, whereas eluates from A1 or Tn cells react with A1, Tn and Cad cells.


RB, a determinant defined by lymphocytotoxicity being associated with ABO blood groups and ABH secretor status. Further evidence that RB is not produced by lymphocytes. In a case of chimaeric male twins which showed red and white cell chimaerism, the latter being demonstrated using polymorphism of autosomes, it could be shown that the determinant RB, being defined by lymphocytotoxicity and associated with ABO blood groups and ABH secretor status, is probably absorbed from the serum on to the lymphocyte surface and not produced by the cells themselves.


Immunochemical investigations on toad (Bufo) eggs: comparative studies on three species (B. bufo, B. viridis, B. calamita). Eggs of three Bufo species (B. bufo, B. viridis and B. calamita) were examined for blood group activity. B. bufo showed distinct A activity, whereas in B. viridis and B. calamita a marked H-activity was observed. These results correspond with the zoological systematic classification of the three toad species tested.


Hair growth inhibition as a method of screening drugs for local antimitotic activity. The intradermal injection of certain drugs with antimitotic properties in the guinea pig resulted in localized areas of reversible hair loss or hair growth inhibition. The size of the affected field was related to the dose. This may provide a useful and simple method for the rapid screening of potentially useful agents for local cytostatic activity.


Cross-antigenicity and immunogenicity between capsular polysaccharides of group C Neisseria meningitidis and of Escherichia coli K92. Antibodies to capsular polysaccharides of group C Neisseria meningitidis are often found in sera of young adults despite infrequent nasopharyngeal carriage and low rate of attack of N. meningitidis in the United States. Thus, experiments were designed for detection of bacteria cross-reactive with N. meningitidis. Among 3,264 cultures of stool, urine, blood, and cerebrospinal fluid, only 14 strains were found to be cross-reactive; all were Escherichia coli possessing the K92 capsular polysaccharide. The somatic O-antigens were 16, 13, 23, and 73; the flagellar antigens were H4 and 34. All K92 strains of E. coli showed the expected fermentations, were sensitive to common antibiotics, and lacked enteropathogenicity. Antigens of both E. coli K92 and group C N. meningitidis are capsular, acidic polysaccharides composed of sialic acid. The K92 polysaccharide is N- but not O-acetylated, sensitive to neuraminidase, and linked by alpha-2,8- alternating with alpha-2,9-ketosidic bonds. The K92 polysaccharides from all E. coli studied had similar biophysical and immunological properties. Intravenous injection of formalin-treated K92 organisms induced precipitating and bactericidal antibodies to polysaccharides of N. meningitidis. E. coli K92 strains may provide an alternative immunogen for prophylaxis against disease due to group C N. meningitidis in infants and young children.


Neutral proteases of human granulocytes. IV. Interaction between human granulocyte collagenase and plasma protease inhibitors. Purified human granulocyte collagenase was inactivated by serum through the formation of complexes with alpha 1-antitrypsin and alpha 2-macroglobulin. A molar combining ratio of 1:1 was observed for each inhibitor. The affinity of alpha 2-macroglobulin was about 10 times that of alpha 1-antitrypsin for granulocyte collagenase. The molar concentration of alpha 1-antitrypsin in the blood exceeds that of alpha 2-macroglobulin by about 12 times, so that the inhibitors may be equally important for defence against granulocyte collagenase.


Pharmacokinetic behaviour of bleomycin-cobalt-57 with special regard to intraarterial perfusion of the maxillo-facial region. Bleomycin was traced by the isotope Cobalt-57. By using this radioactive substance the pharmacokinetic behaviour of the drug was examined. It was given either intravenously or intra-arterially or directly into the tumor in patients suffering from epidermoid carcinoma in the maxillo-facial region. The differences in the pharmacokinetic data are delineated. Practically, the intra-arterial route of administration is the best one because of the high adherence rate of the drug to the perfused tissue.


Dual effects of concealed A-V nodal conduction in man. An interpolated premature ventricular contraction (PVC) may produce either complete block of the next sinus impulse or depression of A-V nodal conduction with a prolonged A-H interval. When a PVC results in partial depression of a A-V nodal conduction, the effect on subsequent premature atrial stimuli is unknown. The authors have recently observed three patients in which the effect of a premature ventricular stimulus with interpolation on the functional refractory period of the A-V node could be measured. In case one an interpolated PVC sufficient to prolong the A-H interval from 80 to 120 msec was followed by programmed premature atrial stimuli which resulted in no additional A-V nodal delay, and the apparent functional refractory period of the A-V node was reduced from 420 to 330 msec when compared with the atrial extrastimulus technique. In case two a programmed ventricular extrastimulus prolonged the A-H interval in the following sinus beat from 120 to 240 msec; atrial extrastimuli then resulted in only minimal increments in A-V nodal delay and the apparent functional refractory period of the A-V node was reduced from 590 msec. A ventricular extrastimulus in case three increased the resting A-H interval from 60 to 115 msec; conduction of atrial extrastimuli then resulted in a reduction in the functional refractory period of the A-V node from 465 to 400 msec. In each case an interpolated premature ventricular stimulus produced (1) depression of A-V nodal conduction in the ensuing sinus beat A1 and (2) relative facilitation of A-V nodal conduction of a subsequent premature atrial stimulus (A2). The functional refractory period of the A-V node was reduced when compared with the atrial extrastimulus technique alone.


T-lymphocyte-enriched murine peritoneal exudate cells. IV. Genetic control of cross-stimulation at the T-cell level. Antibodies raised against many structurally related antigens have been shown to cross-react extensively. Manifestations of T-cell immunity, on the other hand, appear to be more restricted in their ability to be elicited by cross-reacting antigens, although examples have been reported. This paper explores the nature of the cross-reactions at the T-cell level among the branched-chain copolymers (T,G)-A--L, (phi,G)-A--L, (H,G)-A--L, and G-A--L, as well as a related linear terpolymer, GAT, in a variety of mouse strains using the peritoneal exudate T-lymphocyte-enriched cells (PETLES) proliferation assay. (T,G)-A--L, (phi,G)-A--L, and GAT could cross-stimulate cells immune to the other two antigens, whereas (H,G)-A--L, (T,G)-Pro--L, and G-A--L showed no cross-stimulations. The extent of the cross-reactions varied with the mouse strain and was shown to be under the control of immune response genes. It was necessary for the strain to be able to respond to both the immunogen and the cross-reacting antigen, when used as an immunogen, in order for cross-stimulation to occur; however, this was not always sufficient. Several examples of unequal or one-way cross-reactions were found. In addition, the immune responses to (H,G)-A--L and (phi,G)-A--L showed no cross-reactions with the other antigen even though their Ir genes were both mapped to the K region or I-A subregion. The problem of accounting for such fine specificity of T-cell recognition in lieu of the genetic evidence demonstrating only Ir gene control of the response is discussed.


Hapten-specific hemolytic plaque assays usually fail to detect most of the diversity in the anti-hapten response. Immunization of rabbits or mice with a single, chemically defined hapten elicits populations of plaque-forming cells (PFC) detectable not only on sheep erythrocytes (SRBC) bearing the immunizing hapten, but also on SRBC bearing structural analogues of the immunizing hapten. Most of these analogue-reactive PFC preferentially lyse analogue-conjugated SRBC and cannot be detected on erythrocytes bearing the immunizing hapten. Thus, they represent heretofore largely unstudied components of the secretory B-cell response to haptenic immunization, and they have been termed alloreactive PFC. Such alloreactive PFC are detectable using either classical small haptens or tripeptide-enlarged counterparts of these classical haptens. They are present in large numbers both in direct and in indirect PFC assays, and they are elicited in response to both thymic-dependent and thymic-independent antigens. Relatively few alloreactive PFC can be attributed to cells producing hapten-carrier or "bridge area"-specific antibodies. Since the antibodies released by alloreactive PFC can also be detected by passive hemagglutination, their presence does not appear attributable to vagaries of complement activation. Numerous coexisting alloreactive PFC populations are detectable after haptenic immunization. In early direct PFC responses it is not nucommon for a single alloreactive PFC population to outnumber the population of PFC detectable on SRBC bearing the actual immunizing hapten. These alloreactive PFC may be the source of at least some of the new "nonspecific" Ig which is formed at the time of immunization but about which little is known for lack of available techniques. Some possible implications of these findings on the specificity of B precursor cell activation are discussed.


Virus-replicating T cells in the immune response of mice. I. Virus plaque assay of the lymphocytes reactive to sheep erythrocytes. Virus plaque-forming cell assay with vesicular stomatitis virus (VSV), which had been originally introduced by Bloom and his colleagues as a tool for the enumeration of activated lymphocytes, was first applied to the immune response of mice to a widely used antigen, i.e. sheep red blood cells (SRBC). When spleen cells taken from mice previously primed with SRBC were cultured in the presence of the antigen, lymphocytes capable of replicating VSV (antigen-induced virus plaque-forming cells, Ag-V-PFC) were generated in the culture. They seemed to appear as early as 1 day of culture, and the peak was attained by the 2nd day. Most of Ag-V-PFC belonged to T-cell population, since 80-90% of Ag-V-PFC was killed by the treatment of cultured cells with anti-thymocyte serum plus complement. In vitro generation of Ag-V-PFC seemed to be highly cross-reactive (about 40%) with a related antigen (horse red blood cells). Ag-V-PFC detected in the present experiment may not represent helper T cells, effector T cells, or their precursors because of the following: (a) The generation of Ag-V-PFC was completely suppressed by the addition of anti-SRBC mouse serum in the culture, though the helper activity was apparently augmented by the same treatment. (b) Development of Ag-V-PFC was almost completely suppressed by the pretreatment of mice with cyclophosphamide 2 days before immunization, by which delayed-type hypersensitivity (DTH) was markedly augmented. (c) After the immunization of mice, Ag-V-PFC began to develop just when the level of DTH declined, at which point helper activity of the spleen cells also diminished. A possible role of Ag-V-PFC in the immune response was discussed.


Immunochemical evidence for an additional H-2 region closely linked to H-2D. Anti-H-2 reagents have been tested on solubilized spleen cell preparations in combinations expected to be specific for D region products. Two different types of molecules were detected. One showed the expected reactivity with both antisera to private and antisera to public specificities. However, an additional molecule was detected which reacted only with antisera to public specificities. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis migration patterns indicated that both products have a similar molecular size of approximately 45,000 daltons. The data therefore present chemical evidence for the existence of a third H-2-associated gene product of 45,000 mol wt in addition to the classical H-2K and H-2D antigens.


Mechanism of compact-colony formation by strains of Staphylococcus aureus in serum soft agar. Compact-colony forming active substance (CCFAS), the material responsible for the compact colonies of Staphylococcus aureus observed in serum soft agar, was found to be an alkaline-stable, associated polysaccharide containing galactose, N-acetylglucosamine, ribitol, phosphorus and a small quantity of alanine. This substance, when extracted from strains unable to produce protein A clumping factor, was able to absorb the serum-reacting factor whereas a teichoic acid preparation of one strain could not. The formation of CCFAS was unaffected by the age of the cells, whereas when staphylococci were cultured at alkaline pH, young cells produced more clumping factor than old ones. Both fibrinogen and its degradation products were capable of inducing compact colonies in a strain of S. aureus. The ability of human sera to interact in compact-colony formation was independent of the immunoglobin content. Thus neither protein A, clumping factor, nor teichoic acid participate in the CCFAS reaction.


Protargol impregnation of plastic-embedded semi-thin sections: a simple method to select degenerative areas for electron microscopy. 0.5 mu plastic-embedded sections, obtained from aldehyde-osmium fixed rat and cat spinal cord, were impregnated in a 0.5% Protargol solution for 24 hours at 56 degrees C. Reduction was performed in a sodium sulphite- (5%) hydroquinone (1%) developer. Terminals undergoing Wallerian degeneration stand out as easily discernible black dots; corresponding osmiophilic degenerative patterns are demonstrated in consecutive thin electron microscopic sections. This simple technique enables successful trimming of blocks to obtain areas with the highest frequency of terminal degeneration.


Serum globulin changes in patients with craniocerebral trauma. Investigations of serum total protein, albumin, alpha-1, alpha-2, alpha-2M, beta- and gamma-globulin changes are reported in 48 patients with craniocerebral trauma. Only alpha-2 and alpha-2M globulins showed important variations, the first rising to three to four times normal values (112% on average) and directly correlating with the amount of tissue lesions. Alpha-2M changes were rather irregular with a tendency to severe decrease during the first days in patients with a bad prognosis. Reasons for non-regular changes of alpha-2M are discussed. Alpha-1 globulin showed a similar though less marked increase. Gamma globulin, on average, was diminished in the first week.


Electron microscopic observations of the mechanisms of terminal club formation in transected spinal cord axons. Following transection of spinal cords, axoplasmic flow occurred in the axons both proximal and distal to the transection. In the proximal axonal stump, the direction of flow was proximo-distal whereas in the distal axonal stump the direction of flow was disto-proximal. Thus the flows in transected axons were all directed toward the point of transection. Electron microscopic observation of the spinal cord tissue bordering at the cut ends of spinal cord stumps showed various structural changes in the axons and myelin sheaths of the transected fibers near the cut ends which interfered with the axoplasmic flow leading to the formation of the terminal clubs. Five prototypes of changes in axons and myelin sheaths near the cut ends of fibers are described and the mechanism of terminal club formation is discussed. It seemed that the terminal clubs within the transected spinal cord stumps rere of the axoplasmic flow toward the cut ends of the fibers, and (2) interference by structural changes which developed within individual fibers consequent to spinal cord injury.


Erythrocyte-UFA (Eufa) mobility test for multiple sclerosis: implications for pathogenesis and handling of the disease. Erythrocytes from patients with Multiple Sclerosis (MS) show a highly significant reduction in their absolute electrophoretic mobility in the presence of linoleic and arachidonic acids (LA; AA). Patients with other (destructive) neurological disease (OND) and normal subjects show an increased absolute mobility of their erythrocytes in the presence of LA and AA. About 40 per cent of blood relatives of MS patients show an intermediate type of reaction - being slowed by LA and speeded up by AA. Administration of LA (or gamma linolenate) to an MS patient for some months leads to change in the mobilities from the MS to normal type, the AA result altering first. The effect of LA and AA on the absolute mobility of RBC may thus be used as a simple laboratory test involving a long established technique and eliminating the animal and other needs of the macrophage electrophoretic mobility (MEM) test. The implications of these findings for our understanding and handling of MS are briefly discussed.


The perifascicular atrophy factor. An aid in the histological diagnosis of polymyositis. 19 biopsies of polymyositis patients were compared with 19 matched controls. The presence of smaller fibres in the periphery of the fascicles has been analyzed quantitatively using a perifascicular atrophy factor. The thinner fibres are multiplied by a factor from 1-4, considering their significance for the diagnosis of fibre atrophy. The value obtained with this method from centrally located fibres as related to the value from peripherally located ones is called the perifascicular atrophy factor. If this is less than -300 a myopathy of the group of the polymyositis/dermatomyositis can be assumed. 47 per cent of dermatomyositis biopsies and none of the controls were below this range


Specific staining of the axon membrane at nodes of Ranvier with ferric ion and ferrocyanide. Ferric ion and ferrocyanide were used as stains for light and electron microscopy of peripheral nerves. In rat sciatic nerves, it was found that ferric ion preferntially binds to the cytoplasmic surface of the axon membrane at nodes of Ranvier but not at internodal regions. In myelinated axons in the electric organ of the gymnotid fish, Sternarchus albifrons, the small excitable nodes are similarly stained, but the larger inexcitable nodes are not stained by ferric ion. Staining of the inner surface of the nodal membrane appears to be related to a structural specialization of this membrane, rather than accessibility to stain. Our data thus show a chemical differentiation of the inner surface of the axon membrane between nodes and internodes in normal peripheral nerve fibers and between the inner surface of the axon membrane at active nodes, and the internodes in the Sternarchus electrocyte axons.


Toxic polyneuropathies after sniffing a glue thinner. In West Berlin in the autumn of 1975 through the following 5 months we observed 18 juvenile patients who had a toxic polyneuropathy and had sniffed a glue thinner. The neurological picture consisted of a symmetrical, progressive, ascending, mainly motor, polyneuropathy with pronounced muscle atrophy and characteristic vegetative alterations. The height of the disease was reached after 1 1/2-2 1/2 months and was characterized by tetraplegia in 7 patients. After 8 months all patients still had a motor deficit. Nerve biopsy showed paranodal axon swelling, dense masses of neurofilaments and secondary myelin retraction. The neurological and morphological data correspond to the "glue sniffer's neuropathy" and the n-hexane and MBK polyneuropathy after industrial exposure, as described in 10 cases to date. However, there was no MBK in the glue thinner. The polyneuropathies occurred in close time relation with the denaturation of the thinner with MEK (2-butanone). It is concluded from the data n-hexane and MBK have a common toxic mechanism with primary axonal changes and that there is an additional synergistic effect of MEK.


[Global aphasia. The clinical picture and a consideration of the neurolinguistic structure (author's transl)]. Global aphasia is described as a unitary syndrome which is characterized by a severe impairment of all linguistic capabilities. Speech production is extremely limited and consists of stereotyped phrases, recurring utterances or a few isolated words which are usually neologistically distorted. The patients are unable to express their thoughts in a situationally adequate manner. Language comprehension is restricted to simple questions and commands. A clinical description of the syndrome is given and the neurolinguistic structure of the disorders is discussed.


Aseptic meningitis: frequency among Israeli ethnic groups. The relative frequency of aseptic meningoencephalitis (AME) was compared in populations of diverse origin, A countrywide search of Israel during 1969-1970 disclosed 1350 cases who fit strict diagnostic criteria. The average annual incidence was 21.6 per 100000 population. The total incidence was similar in Afro-Asian, Euro-American and Israeli Jewish groups but among Israeli Arabs, the incidence was apparently lower. Age-specific incidence showed a peak in infants under one year of age among Arabs and Afro-Asian Jews whereas Euro-Americans and Israeli Jews had a peak incidence at 5-9 years. Larger family size among Arabs and Afro-Asian Jews might account for the higher incidence in infants. Age-specific incidence may be a better index than total incidence of important differences in AME among various ethnic groups.


Autoradiographic demonstration of proliferating cells in cerebrospinal fluid. The proliferative activity of cells, isolated from 82 human CSF specimens, was examined by 3H-thymidine autoradiography. High labelling indices (LI) were found in acute viral meningitis (up to 8 per cent) and radiculitis (up to 6 per cent). CSF cell proliferation was also shown in the subacute stages of viral diseases and in other inflammatory processes (LI ranging from 0.5 per cent to 3 per cent). Most of the cells labelled from these CSF specimens were large lymphocytes, "lymphoid cells" and plasmacytes. Their presence in CSF is presumed to indicate an immune reaction. By the demonstration of a proliferative activity of these cells, aseptic inflammatory processes can be differentiated from "unspecific" pleocytosis. Because of a correlation between the LI of CSF cells and the stages of some inflammations, this method is suggested for an assessment of pregression or remission of chronic processes, e.g. "chronic meningitis" and multiple sclerosis. It can also be used in experimental research: the same type of mononuclear cells was labelled after having been cultured for 23 hours prior to the incubation with 3H-thymidine. Proliferating tumor cells as well as proliferating non-neoplastic mononuclear cells were demonstrated in CSF from various neoplastic diseases. In the clinical diagnosis of these processes, the method is of limited value. It proved very useful, however, for an assessment of the therapeutic effects of intrathecal cytostatic therapy. CSF specimens from non-inflammatory and non-neoplastic diseases regularly contained very few proliferating cells (LI: less than 0.1).


Comparison of attitudes and cognitive achievement of nursing students in three instructional strategies. This study is a comparison of the attitudes and the cognitive achievement of nursing students in three instructional strategies (the traditional lecture, black and white televised instruction, and independent color televised instruction via the Dial Access Information Retrieval System). Though only thirty-one percent (31%) of students indicated prior exposure to independent study methology, sixty percent (60%) identified a desire for active involvement in courses of study. The only significant finding regarding attitudes toward the three strategies was a preference for an greater interest in color videotapes than for black and white televised material. No significant differences in cognitive achievement were noted between any of the three strategies. Implications from the study for curriculum revision are discussed, stressing the probable value of maintaining traditional techniques concurrent with innovative methodological experimentation.


New neonatal problems of blood coagulation and fibrinolysis. I. The change of plasmin inhibitor levels in the newborn infant. "Hemorrhage in the newborn" has long been recognized as merely a result of vitamin K deficiency. However, it is also recognized that fibrinolysis, especially the correlation between the plasminogen-activator and plasmin-inhibitors, play an important role in this disease during the neonatal period. With this in mind, we compared thromboelastograms (TEG) from samples with and without urokinase (plasminogen-activator). In 13 out of 15 newborn infant blood-samples (prior to and after addition of urokinase) the thromboelastogram showed the pattern of a consumption coagulopathy. The change in the concentration of plasmin-inhibitor during the neonatal period was also measured using alpha2-macroglobulin, alpha1-antitrypsin and antithrombin III with M-partigen-plates. The value of alpha2-macroglobulin showed normal adult levels but the value of alpha1-antitrypsin and antithrombin III did not even reach half of the adult level. During the newborn period, the plasmin-inhibitor shows a remarkable lowering tendency and it may be surmised that with such a lowering tendency plasmin-inhibitor may constitute an exceptionally large handicap when the activator is working. This is especially true in the case of lung hemorrhage since the activator arises from a severe pathological state in the lungs and in addition because this is complicated by the lowering of plasmin-inhibitor. These results indicate that the low level of plasmin-inhibitors work synergistically with the high value of activator. The low level of antithrombin III could be the reason for coagulation disorders such as disseminated intravascular coagulation, (DIC).


Serum alpha2-macroglobulin levels in tuberose sclerosis. The serum levels of alpha2-macroglobulin (alpha2-MG) were determined by radial diffusion if fifty-four cases with tuberose sclerosis and compared with forty-seven institutionalised control subjects of similar age and sex distribution. Although higher levels of alpha2-MG were found in the females of both groups when compared with the males, this increase was not significant. The tuberose sclerosis subjects showed consistently elevated levels of alpha2-MG when compared to the control group for both the males and females separately and combined. With the two sexes combined this elevation was significant at p less than 0.001. The significance of this observation has been discussed both from the point of view of the possible mechanism involved and the use of this estimation in genetical counselling.


Plasma acute-phase reactant proteins in tuberose sclerosis. The quantitative levels of seven of the acute-phase reactant proteins were measured in the plasma of a sample of fifty-four adults all diagnosed as suffering from tuberose sclerosis and compared to that derived from a group of control subjects resident in a hospital for the mentally subnormal. Abnormal levels were found in one or more of the proteins in all but thirteen cases, with no fewer than twenty-six subject (forty-eight per cent) showing results outside the 2SD limits in three or more of the parameters investigated. Only in the alpha1-antitrypsin and fibrinogen were the means not significantly different from the control means but even in these two proteins ten and seven cases respectively individually exceeded the normal mean by more than 2SD. It is suggested that these observations can be explained as the normal response to the presence of neoplastic tissue, and the routine investigation of these and other biochemical components known to respond to the presence of neoplastic tissue may be of help in genetic counselling.


Contrast enhancement of negatively stained macromolecules and biomembranes by single sideband phase contrast interference. A straightforward procedure is described for the production of contrast enhancement of negatively stained macromolecules and biological membranes by single sideband phase contrast interference (electron optical shadowing). The instrumental adjustment required to produce this type of phase contrast illumination is readily achieved by beam deflection from the strioscopic (dark field) mode. Part of the hollow cone of electrons from the annular condenser aperture that are unscattered by the specimen are permitted to pass through the objective aperture and interfere with the scattered beam. The electron optical shadowing effect is produced because only one side of the unscattered beam is used. Careful adjustment of the beam tilt control, with the ability to tilt in any azimuth, allows optimal illumination conditions to be achieved. The results presented show the increased image contrast obtained using as specimens the purified cylindrical macromolecule from human erythrocyte membranes, purified nuclear envelopes and collagen fibres.


The influence of the embedding medium when staining for electron microscopy: the penetration of stains into plastic sections. Using a sectioned-section procedure it was found that only a few structures, e.g. the connective tissue elements, were deeply penetrated by phosphotungstic acid (PTA) and uranyl acetate (UA). Reynold's lead citrate appeared to penetrate more generally. Using stains selective for the embedding medium, and also observing the surfaces of shadowed sections, it was concluded that structures readily penetrated by polar stains were only poorly infiltrated by non-polar embedding media. It is argued that this differential infiltration leading to differential penetration by polar stains largely controls the pattern of staining of ultrathin sections by PTA and UA.


A new metachromatic stain technique for paraffin-embedded neural tissue using thionin. A metachromatic staining procedure which differentiates Nissl substance in neuron cell bodies, neuropil area, and axonic fibres of passage with distinct colours with no counterstaining has been developed for paraffin-embedded tissue utilizing thionin stain. Frozen sections of neural tissue fixed in formalin may also be satisfactorily stained by this procedure, and both paraffin-embedded and frozen sections have been found to retain the distinct colours for a year with little fading. Since metachromatic staining of paraffin-embedded neural tissue has not been successfully achieved before, the following procedure will be especially valuable for studying small vertebrate brains, or other central nervous tissue which must be processed by the paraffin method.


Human B-lymphocyte antigens expressed by lymphocytic and myelocytic leukemia cells. II. Detection by human anti-B-cell alloantisera. The majority of human lymphocytic and myelocytic leukemia cells express a polymorphic antigen that is found on peripheral blood B-lymphocytes and cultured lymphoblastoid B-cell lines. These B-lymphocyte antigens were detected by 34 human alloantisera that were repeatedly absorbed with pooled platelets to remove all activity against HLA antigens and T-lymphocytes. Absorption studies indicated that a common antigen was present on both B-lymphocytes and positive leukemia cells. Leukemia cells could be subdivided into two groups based on the presence of the B-lymphocyte antigen. Fourteen of 18 acute myelocytic leukemia cells, 10 of 13 acute lymphoblastic leukemia cells, 4 of 6 chronic myelocytic leukemia cells, and 2 of 2 chronic lymphocytic leukemia cells were positive. This group of leukemia cells also reacted with rabbit anti-B-cell sera raised to papain digests of spleen cell membranes. F(ab')2 fragments of the rabbit antsera were shown to specifically block the reactions of the human antisera against B-cells and leukemia cells. These results suggested that the rabbit and human anti-B-cell sera were reacting with identical molecules. This conclusion was supported by immunoprecipitation data.


Effects of streptovaricins and their degradation products on RNA-directed DNA polymerase of Rauscher leukemia virus. The activities of streptovaricin complexes, streptovaricins, streptovals, and streptovarinic degradation products were elevated against RNA-directed DNA polymerases of Rauscher leukemia virus, DNA-dependent DNA polymerase of bacterial and mammalian cells, and DNA-dependent RNA polymerases of mammalian origin. The activities of streptovaricins were also listed for comparison purposes. The effects of streptovaricin complexes on viral DNA polymerases varied significantly from lot to lot, and streptovaricin complex lot 7 was the most active. All the streptovals and streptovaricin degradation products except varicinal A showed a marked improvement (twofold to tenfold) in activity against the viral enzyme over the parent streptovaricins. None of these compounds, however, displayed any significant effect on either the DNA polymerase of L1210 leukemia cells and Escherichia coli or the RNA polymerase of isolated nuclei of mouse liver. As a result of tests in these systems, some specific inhibitors of RNA-directed DNA polymerases of Rauscher leukemia virus were selected.


Effects of streptovaricins and their degradation products on infectivity of Rauscher leukemia virus. The virucidal effects of streptovaricin (Sv) A, SvC, SvD, streptoval (Sval) C, Sval Fc, and streptovarone were evaluated by incubation of the drug with Rauscher leukemia virus (RLV) at 37 degrees C for 60 minutes prior to dillution and addition to cells (in vitro assay) or before ip injection into animals (in vivo assay). The in vitro and in vivo assays were plaque formation and splenomegaly, respectively. A dose-related effect was observed with all six compounds with the in vitro assay. On an equimolar basis, the Sv degradation products, i.e., Sval C, Sval Fc, and streptovarone were most inhibitory, followed by SvD; SvA and SvC were least active. At 0.0625 mumoles, the three Sv degradation products inactivated over 90% of the RLV. Similar results were obtained through the in vivo assay. At 0.06 mumoles, streptovarone, Sval C, and SvD showed 78,62, and 29% inhibition of splenomegaly, respectively; SvA and SvC were essentially inactive. A direct relationship was observed between inhibition on RNA-directed DNA polymrase of RLV by these compounds and their virucidal effects. No drug given at the time of injection, however, showed any significant effect on virus infective processes in vitro or in vivo. The reason for the lack of therapeutic effects of these compounds is discussed.


Antigenic similarity between simian virus 40-induced surface and fetal antigens in hamster. The tumor-associated cell-surface antigen (TSSA) on simian virus 40 (SV40)-transformed hamster cells was studied serologically by a complement-dependent cytotoxicity test. An antiserum was obtained from guinea pigs inoculated with SV40-transformed hamster cells. The serum was cytotoxic to SV40-transformed hamster cells after absorption with 15-day hamster embryo cells, hamster cells transformed either by polyoma virus or adenovirus 12, various tissues of hamster origin (brainliver, spleen, and kidney), or sheep red blood cells. These results indicated that the major TSSA induced specifically by SV40 was similar or identical to the antigen present during early stages of embryogenesis.


Effect of serum deprivation on myelinating mouse cerebellum cultures. In order to investigate the effect of nutrient deprivation on brain development, mouse cerebellum cultures were grown in nutrient media with reduced serum concentration. After 9--18 days in vitro, experimental cultures grown in media with low serum concentration exhibited a delayed and retarded myelination. Electron microscopic examination of experimental cultures revealed profiles of deficient myelination, but showed normal neuronal and synaptic structures. The amount of myelin basic protein measured by radioimmunoassay was markedly reduced in experimental cultures. Activity of cholinesterase in these experimental cultures was also decreased.


Interspecies determinants of Friend leukemia virus antigens involved in cytolysis of virus-pfoducing cells. The cytolytic reactivity of a complex goat anti-feline leukemia virus (FeLV) antiserum for mouse cells (Eveline) releasing large quantities of Friend leukemia virus (FLV) was analyzed by the sensitive [14C]nicotinamide release microcytotoxicity assay. Whereas this interspecies killing reactivity could be blocked by absorption of the goat anti-FeLV serum with a preparation of disrupted FLV, absorption with purified FLV gp71, the major envelope glycoprotein, had no effect. Subsequent serum absorptions with the individual FLV structural polypeptides revealed that the lysis of the Eveline cells by the goat anti-FeLV serum is mediated by antibodies recognizing the interspecies determinant of p30, the major internal capsid protein. The expression of this internal viral component at the surface of virus-producing cells is discussed further. The results also demonstrated that removal of approximately 70% of the carbohydrate portion of gp71 with a preparation of glycosidases did not affect the integrity of its interspecies determinant; these results are in agreement with an earlier study (Bolgnesi et al., 1975) that examined primarily the group- and type-specific sites.


Increased length of DNA made by virions of murine leukemia virus at limiting magnesium ion concentration. Conditions have been developed for reverse transcription by detergent-disrupted virions of Moloney murine leukemia virus which permit synthesis of molecules that appear to be complete transcripts of the 35S RNA subunits. At limiting Mg2+ concentration, DNA is synthesized in good yield, up to a maximum size of about 2.4 X 10(6) daltons. DNA larger than 2 X 10(6) daltons, taken from alkaline sucrose gradients, has no detectable self-complementarity and was protected from digestion by S1 nuclease to an extent of 90% by annealing to 70S RNA. All size classes of DNA made in these reactions are primed with RNA, because all are initiated with a pApdAjunction. To produce such long molecules, it is necessary to keep the concentration of Mg2+ in the reaction mixture below the total concentration of deoxyribonucleoside triphosphates. Under these conditions, degradation of the RNA template is minimized. The rate of DNA synthesis is also slowed by 30 to 50%, but products longer than 5,000 nucleotides, which are not found otherwise, are completed between 3 and 6h of reaction.


Common structural antigen of papovaviruses of the simian virus 40-polyoma subgroup. An antigenic determinant common to the major capsid polypeptide (VP1) of simian virus 40 (SV40) and polyoma virus is described. Antisera prepared against intact viral particles reacted only with cells infected with the homologous virus by immunofluorescence tests (IF). However, antisera prepared against disrupted SV40 particles reacted in IF with both polyoma- and SV40-infected permissive cells. The cross-reaction with polyoma was localized to VP1 by the following evidence. (i) The IF cross-reaction was inhibited by preincubation of the antiserum with purified SV40 VP1; (ii) purified radiolabeled polyoma VP1 was precipitated by the cross-reactive serum, and this reaction was inhibited by unlabeled SV40 VP1; (iii) other antisera prepared against purified SV40 VP1 or polyoma VP1 reacted in IF with both SV40- and polyma-infected permissive cells. These cross-reacting antisera also reacted in IF with permissive cells infected with BK virus, rabbit kidney vacuolating virus, and the stumptailed macaque virus, suggesting that all members of the polyoma-SV40 subgroup share a common antigenic determinant located in their major capsid polypeptides.


Expression of an early, nonstructural antigen of herpes simplex virus in cell transformed in vitro by herpes simplex virus. Hyperimmune rabbit antiserum to an early, nonstructural herpes simplex virus type 2 (HSV-2)-induced polypeptide (VP143) reacted in immunofluorescence tests with a variety of cell lines transformed by HSV-2. Cytoplasmic fluorescence was observed in 10 to 50% of HSV-2-transformed cells, whereas no fluorescence was observed in cells transformed by other oncogenic DNA viruses or by a chemical carcinogen. VP143-specific reactivity could be absorbed from anti-VP143 serum with HSV-2-transformed cells but not with cells transformed by other agents. When HSV-2-transformed cells were synchronized in mitosis and examined at various times postmitosis for VP143-specific fluorescence, the expression of VP143 was shown to be cell cycle dependent.


Viral protein synthesis in Friend erythroleukemia cell lines. Viral protein synthesis was studied in two Friend virus-induced erythroleukemia cell lines (Ostertag cell lines FSD1-F4 and B8) by the technique of immuno-precipitation with monospecific antisera to the major envelope glycoprotein gp70 and major core protein p30. One of the cell lines (F4) releases active Friend virus complex to the growth medium, where release of virus from the other cell line (B8) is barely or nondetectable. It was found that in the nonproducer cell line B8, a large-molecular-weight protein of about 65,000 containing p30 antigenic determinants is synthesized, yet no p30 is produced upon prolonged incubation and chase, suggesting that this might be the actual lesion that prevents mature virus production by these cells. In both cell lines, the predominant protein species that is immunoprecipitated with monospecific anti-gp70 serum is a protein of 55,000 to 60,000 daltons that is labeled with glucosamine to a much lesser extent that gp70 and appears to become heterogeneous with time. Large amounts of gp70 can be detected in the cell-free medium, but none of the unstable species of 55,00 to 60,000 molecular weight.


Purification and molecular characterization of adenovirus type 2 DNA-binding protein. An adenovirus type 2 (Ad2) DNA-binding protein was purified by sequential DNA-cellulose, Sephadex G-200, and DEAE-Sephadex chromatography, with a yield of 120 mug of binding protein (95 to 99% homogeneity) starting with 2 X 10(9) infected cells. By omitting the Sephadex G-200 step, 400 to 600 mug of 95% pure binding protein was obtained. To obtain high yields of highly purified binding protein, it was necessary to include deoxycholate and Nonidet P-40 at selected stages during the preparation. The highly purified binding protein appeared to have retained its native stage as indicated by: (i) binding to single-stranded but not native Ad2 DNA, (ii) almost complete precipitation by immunoglobulin G from hamsters immunized by extracts of tumors induced by Ad2-simian virus 40 hybrid viruses, and (iii) identical sedimentation coefficient with binding protein obtained from DNA-cellulose chromatography only. Zonal centrifugation in sucrose gradients and gel filtration revealed that purified binding protein has a sedimentation coefficient of 3.4S and a Stokes radius of 5.2 nm. Based on these two values, a molecular weight of 73,000 was calculated, in agreement with the estimate from sodium dodecyl sulfate-polyacrylamide gel electrophoresis. A frictional ratio of 1.88 was calculated, suggesting that the Ad2 DNA-binding protein does not have a typical globular protein structure.


The changing role of chemotherapy in treatment of non-seminomatous germinal testicular cancer. Although this series is too small for meaningful statistical conclusions it demonstrates the effectiveness and hazards of bleomycin and vinblastine combined with other treatment modalities in the handling of non-seminomatous germinal testicular cancer. Bleomycin and vinblastine therapy is indicated 1) as a last restor in patients who have failed with other forms of therapy, 2) to convert high stage lesions to a lower stage and 3) as adjuvant therapy in the lowest stage lesions. The indications, risks and alternatives to chemotherapy, especially in patients with low stage tumors, must be realized by the physician and patient prior to its administration. However, the results reported here and elsewhere are so encouraging that we strongly recommend more aggressive use of these agents in any treatment regimen involving non-seminomatous tumors.


Evaluation of research needs in nephrology in urology. Obstructive and neuromuscular disorders affecting the urinary system. Obstructive and neuromuscular disorders are common problems of the urinary tract. They cause considerable morbidity and some mortality. In all problem areas research efforts have been and continue to be limited. The principal reason for the lack of research efforts in these areas is the lack of trained established investigators interested in the problems of the urinary tract. Unfortunately, most basic scientists with the research and investigative background to solve many of these problems see them as mundane and not worthy of investigation. In addition, these same basic scientists are out of touch with clinical urologic diseases and probably do not appreciate the magnitude of the problems. In our opinion the solution is 2-fold: to train qualified investigators in urology and nephrology, and to interest, educate and collaborate with the basic scientists about these clinical problems of the urinary tract.


[Bathmotropic effect of panangin and strophanthin in patients with extrasystole]. The antiarrhythmic effect of Panangine and Strophantin was studied by way of rhythmography in patients with extrasystole of different etiology. The strophantin (0.25 mg) an Panangine (2 vials intravenously) tests, and therapeutic courses of Panagine (2 coated tablets 3 times a day for 5--7 days) or of Strophantin (0.25 mg) with Panagine (2 vials intravenously) for 7--10 days were conducted. A course of Strophantin with Panangine was given only to those patient who did not increase the number of their extrasystoles after the Strophantin test. Such a course resulted in the most striking antiarrhythmic effect (60.3% of the patients).


[The vulnerability of the right atrium. II. Correlations between vulnerability and the atrial refractory periods (author's transl)]. The vulnerability of the right atrium to single electrical stimuli was investigated in 100 non-selected patients. According to the response pattern to extrasystolic stimulation of the right atrium during the relative refractory period, the patients could be divided in 3 groups: Group A: no signs of vulnerability. Group B: 1 to 5 additional atrial extrasystoles after the premature atrial stimulus. Group C: runs of atrial flutter or fibrillation for at least 8 s. The 3 groups were compared with respect to the duration of the effective and relative refractory period of the right atrium. From group A to group C the effective refractory period showed a tendency to shorten, while the relative refractory period showed a tendency to prolong. These results are interpreted as supporting a reentry as the underlying mechanism of the here described atrial vulnerability.


Myocardial ischaemia, risk factors and death from coronary heart-disease. Mortality follow-up is now complete for 5 years in the 18 403 male civil servants aged 40-64 who were examined between 1967-69 in the Whitehall Study of British civil servants. During this period, 277 of them died of coronary heart-disease (C.H.D.); half of these deaths were in subjects in whom the findings at initial screening had suggested early myocardial ischaemia (angina or history of possible infarction according to standard questionnaire, or electrocardiographic evidence of ischaemia). The finding of suspect ischaemia had greater predictive power than the "primary" coronary risk factors, from which it was generally independent. At each level of the primary risk factors, the risk of death from C.H.D. was much greater in the presence of suspect ischaemia; and, with the possible exceptions of glucose tolerance and physical activity, the main risk factors still operated even at the stage of early ischaemia. These findings have implications for future studies of the effects of intervention.


Carboxyhaemoglobin levels in smokers of filter and plain cigarettes. Blood-carboxyhaemoglobin (COHb) levels were studied in 343 healthy male cigarette smokers aged 35-64. The mean COHb level was 30% higher in the 248 men who smoked unventilated filter cigarettes than in the 41 men who smoked plain cigarettes, after adjusting for the number of cigarettes smoked (P less than 0.001). This result was consistent with the carbon-monoxide (CO) yields of these cigarettes: on average, the unventilated filter cigarettes had yields 25% higher than the plain ones. The mean COHb level was 7% higher in the 54 men who smoked ventilated cigarettes than in men who smoked plain cigarettes. Although this difference was not statistically significant, it was in the opposite direction to the result which might have been expected on the basis of the CO yields of these cigarettes: on average the ventilated filter cigarettes had yields 21% lower than the plain ones. The medical implications of these results are uncertain. COHb levels of 3-4% or more can exacerbate angina pectoris and intermittent claudication, and it is, therefore, of concern that COHb levels are higher in smokers of filter cigarettes than in smokers of plain cigarettes. However, any assessment of risk associated with either type of cigarette should take account of the observation from other studies that filter cigarettes are associated with a lower mortality from lung cancer than are plain cigarettes.


Direct evidence for synthesis of valine in man. Plasma valine and 13C-valine concentrations were measured in 2 healthy volunteers and 2 uraemic patients during and after a 3-hour intravenous infusion of the 13-c-labelled alpha-keto-acid analogue of valine. Plasma-valine increased by 23-43%. 13C-valine accounted for most of the early increase, but for less than 30% of the increase 1 hour after infusion. It was concluded that valine had increased by 2 mechanisms with different time courses. Initially, transamination of the infused ketoacid predominated, confirming directly for the first time that the essential aminoacid valine can be synthesised in-vivo both in health and uraemia. The previously unsuspected second mechanism was quantitatively more important but slower in ooperation; it may prove to be the key to understanding the metabolic effects of essential aminoacid precursors.


Cell electrophoretic analysis of lymphocytes and polymorphonuclear cells from patients with rheumatoid arthritis. Cell electrophoresis was performed on peripheral-blood lymphocytes obtained from 11 patients with rheumatoid arthritis (R.A.) and 16 normal controls. On the basis of cell frequency/mobility histograms each subject's lymphocyte electrophoretic profile was resolved into a bimodal distribution of fast and slow cells. The mean electrophoretic mobility (E.P.M.) of the fast-cell population was similar in controls and in R.A. patients but the mean E.P.M. of the slow population was significantly slower in R.A. than in the controls. In the R.A. patients the E.P.M. of the polymorphonuclear cell population was significantly slower than that of P.M.N. cells in the peripheral blood of controls.


Blood-flow in incompetent perforating veins. A new pulsed multichannel ultrasonic Doppler instrument was used to determine the relative blood-flow in the perforating veins of 16 patients with symptoms attributable to their leg venous systems. 9 patients had leg ulceration and 7 did not. 8 (89%) of the ulcer patients had sustained blood-flow from the deep to the superficial veins ("reverse") during calf muscle contraction, compared to 1 (14%) of the group without leg ulcers. This evidence supports the theory that postphlebitic leg ulceration is the result of retrograde flow of blood in incompetent calf perforating veins during walking.


Lipofuscin formation in the developing nervous system of squirrel monkeys consequent to maternal dietary protein deficiency during gestation. The formation of lipofuscin pigment under the extrinsic influence of maternal protein deprivation during most of the gestation period has been demonstrated in the cerebellum and motor cortex of the developing fetuses and neonates in squirrel monkeys. The controls maintained on high protein diets failed to show lipofuscin pigment in appreciable quantities. The dirty yellow to dark brown pigment bodies exist as homogeneous particulates as well as duplex structures, each with a thin, irregular, light brown cortex, studded with one or more darker granules, and a medulla filled with sharp yellow substance. The malnourished neonates, especially in the third and fifth laminae of motor cortex, show loose aggregations of lipofuscin bodies in the apical dendrite and occasionally in the body of the dendritic branches but not in the axon hillock or the body of the axon. Histochemically, the pigment is a complex of lipids, carbohydrates and proteins, showing strong reaction for non-specific esterase and acid phosphatase and mild activity of succinate and lactate dehydrogenase and monoamine oxidase, which puts the pigment observed in the same category as the typical aging pigment described in literature. It is clear that maternal protein deficiency during the critical period of the fetal brain development has an adverse effect on its biochemical maturation. The accumulation of lipofuscin pigment is probably one manifestation of the metabolic changes in the neurons, which may have the impact of hastening the aging process, if protein deprivation is continued into the postnatal period.


Skin test for coeliac disease using a subfraction of gluten. Gluten and various of its fractions and subfractions have been used for intradermal testing in 10 patients with coeliac disease and in 20 healthy control subjects. All the coeliac patients gave positive Arthus-type reactions (type III) to the subfractions of gluten, whereas all the control subjects were negative. The subfraction B2 evoked strong reactions in almost all the coeliac subjects. If tests with larger series confirm the present results, skin testing with B2 may prove to be a useful screening test for coeliac disease. As B2 has already been found to stimulate the lymphocytes of coeliac patients, the present findings are also of interest in relation to the pathogenesis of coeliac disease.


A heparin analogue with specific action on antithrombin III. The effects of heparin and a semi-synthetic heparin analogue were compared in vivo and in vitro. The two drugs differed strikingly in their in-vitro behaviour: unlike heparin, the heparin analogue had little effect in a specific heparin assay or on overall clotting, as measured by the kaolin-cephalin clotting time (K.C.C.T.). When given by parenteral injection, the heparin analogue had almost the same potentiating effect on antithrombin III as mucous heparin, but without a comparable effect on the K.C.C.T. These observations suggest that the heparin analogue may have desirable characteristics for the prophylaxis of venous thrombosis, since it selectively potentiates antithrombin III in vivo while having little effect on overall clotting.


Starvation suppression and refeeding activation of infection. An ecological necessity? The hypothesis is advanced that starvation suppresses and refeeding activates certain infections as an essential part of an ecological balance between man, his animals, and his environment. During famine, then, man fails to thrive, but his ultimate extinction is prevented in part by the parallel decline in fecundity of his "micropredators". In times of plenty the parallel increase in the same predators is a check against his excessive multiplication.


Biochemical markers in human breast cancer. Nineteen biochemical parameters, most of which have been individually advocated as tumour-index-substances for breast cancer, were measured in 51 patients with breast disease, 42 of whom had active breast cancer. Seven of these parameters were raised in more than half of the 17 patients of the series with overt metastases; these were serum ferritin (88%), C-reactive protein (87%), carcinoembryonic antigen (81%), acid glycoprotein (75%), total alkaline phosphatase (64%), sialyl transferase (56%), andthe urinary hydroxyproline/creatinine ratio (73%). The incidence of biochemical abnormalities in patients in this group compared favourably with the results of physical methods of detecting metastases. 7 of 16 further patients without evidence of distant metastases, but who had a poor prognosis as judged by histology of the primary tumour and axillary lymph-nodes, had abnormalities of at least one of the seven parameters. 3 of these patients have relapsed within a year of mastectomy. The results suggest that these biochemical tests could assist in monitoring metastatic disease and could indicate at the time of mastectomy, patients who might benefit from immediate systemic therapy in addition to local treatment of their breast carcinomas.


Hyperventilation: An important cause of pseudoangina. 50 patients initially referred to a cardiac clinic for confirmation or exclusion of angina were found to be habitual hyperventilators. 13 of them also had some organic heart disease. Simple physiotherapy aimed at restoring a normal breathing pattern proved an effective treatment, 76% of those followed for 11-68 months being symptom-free.


Neutrophil function in children who are carriers of hepatitis-B surface antigen. The functional capacity of neutrophils of twenty children who are chronic carriers of hepatitis-B surface antigen (HBsAg) was investigated by histochemical and quantitative nitroblue tetrazolium tests (N.B.T.), an assay of bactericidal capacity against Encherichia coli, and an immunofluorescence test for HBsAg in neutrophils, The histochemical N.B.T. test showed that there was a higher proportion of stimulated neutrophils in chronic HBsAg carriers than in controls, but HBsAg particles were not detected in these cells. Bactericidal activity was significantly reduced in chronic carriers. If the defects in neutrophil function in the HBsAg carrier children preceded their hepatitis-B infections they could have influenced the development of the carrier state. Newborn infants infected with hepatitis-B virus are likely to become chronic carriers and neutrophil functions in uninfected newborn infants are similar to those seen in these carriers of HBsAg.


Mechanism of pancreatic polypeptide release in man. Pancreatic polypeptide (P.P.) is a potent hormonal peptide which has been isolated from the pancreas. Its role in human physiology and pathology is not yet established. After a standard hospital lunch the plasma concentration of P.P. showed a rapid and identical rise in 10 healthy controls, 11 duodenal-ulcer patients, and 6 post-vagotomy patients but remained undetectable in 4 totally pancreatectomised subjects. In contrast plasma-P.P. was unaffected by intravenous administration of glucose, aminoacids, or fat. However, during intravenous infusion of caerulein, a cholecystokinin analogue, P.P. rose by nearly five-fold, and an even greater rise was seen after intravenous injection of Boots secretin. In 19 duodenal-ulcer patients insulin hypoglycaemia produced a rapid rise in plasma-P.P. but this did not occur in any of the 17 patients studied after a truncal vagotomy. Thus P.P. is released by oral but not intravenous nutriments and the existence of an entero-P.P. axis is postulated. One component of this axis may be the vagal innervation but the normal postprandial rise seen after vagotomy suggests that other control mechanisms, such as the intestinal hormones, are more important.


Stress deficiency of the T-lymphocyte system exemplified by Down syndrome. A comparison of immune competence in 26 patients with Down syndrome in an institution and 26 matched healthy controls revealed an atypical pattern of T-cell immunodeficiency in the Down-syndrome patients. The patients with Down syndrome had a lymphocytosis in blood with high counts of T (and B) cells, but with impaired effector function of T cells as judged by anergy to dinitrochlorobenzene, low responsiveness to ubiquitous antigens which elicit delayed-type hypersensitivity reactions, and low mitogenic activity of non-stimulated and phytohaemagglutinin-stimulated lymphocytes in culture. Helper-T-cell function measured by the humoral immune response to flagellin was intact, and there were minor abnormalities of the B-cell system. Attempted restoration of T-cell function with levamisole was unsuccessful. This pattern of T-lymphocytosis with impaired effector function could be explained by "stress-deficiency" of the immune system consequent upon a heavy load of infection in early life.


Vitamin-D in human milk.

Vitamin-D in human milk. The vitamin-D concentration in human milk is reported  to be very low, yet breast-fed infants do not develop rickets. All the earlier assays of vitamin D were made on the lipid fraction of milk, and the aqueous phase was discarded. It is now clear that most of the vitamin D in human milk is present as a water-soluble conjugate of vitamin D with sulphate. The concentration of vitamin-D sulphate in the aqueous fraction of human milk collected at different stages of lactation has been chemically measured. Milk was collected from twenty-two women 3-8 days post partum and fourteen women 4-6 weeks post partum. The vitamin-D sulphate concentration in milk collected between the 3rd and 5th days was 1-78 mug/dl, significantly higher that the 1-00 mug/dl for milk collected between the 6th and 8th days. After the 8th day there was no significant change.


Lymphocyte purine 5'-nucleotidase edficiency in primary hypogammaglobulinaemia. The mean activity of purine 5'-nucleotidase (E.C.3.1.3.5) in lymphocytes was significantly lower in patients with non-familial adult onset "variable" primary hypogammaglobulinaemia than in non-hypogammaglobulinaemic control subjects. These findings suggest that 5'-nucleotidase activity is necessary for normal lymphocyte function. This may be related to its role in facilitating the transfer of newly formed purines across cell membranes by converting them from nucleotides to nucleosides. This enzyme defect may be a characteristic metabolic lesion in at least some of the patients; alternatively, it may be secondary to some other metabolic defect or plasma-membrane change.


Response of lymphocytes from cancer patients to human chorionic gonadotrophin. Positive responses to human chroionic gonadotrophin (H.C.G.) were observed in the macrophage electrophoretic mobility test when lymphocytes from 34/44 patients with various types of cancer and from 7/21 patients with non-malignant disease were tested. Lymphocytes from some pregnant women also responded but those from parous women who were not pregnant did not respond. Positive responses were also obtained with H.C.G. sub-units but not with luteinising hormone.


Duchenne's muscular dystrophy: studies in cultured fibroblasts. Fibroblast cell-lines derived from four patients with clinically confirmed Duchenne's muscular dystrophy were examined by Duchenne's muscular dystrophy were examined by transmission electron microscopy. Cell-lines from patients with Duchenne's muscular dystrophy could be distinguished from normal cell-lines by the presence of characteristic inclusion bodies.


How doctors deal with epilepsy.

How doctors deal with epilepsy. All of those adults with epilepsy on the lists of 17 general practitioners in Metropolitan London were identified. 87% of these agreed to be separately interviewed in their homes by a neurologist and a sociologist. The vast majority had been referred to hospital for investigation, and their hospital notes were inspected. The pattern of care suggests unnecessary referral, unnecessary electroencephalography, inadequate communication of the diagnosis, inadequate medication, and follow-up supervision not related to patient need.


Use of early fetal tissues obtained from suction termination of pregnancy. Organs were identified in the fetal products of pregnancies of short gestation terminated by the suction method. Various tissues from such fetuses were grown in tissue-culture and some of these cultures were used for the isolation of viruses. This new approach would ensure the supply of human embryonic tissue, essential for certain virlogical procedures, as suitable fetuses from pregnancies terminated by hysterotomy become less common. Tissues from these early fetuses could also be used fro transplantation and developmental studies.


An inactive higher-molecular-weight renin in normal subjects and hypertensive patients. Inactive renin which can be converted to an enzymologically active form by acidification to pH 3-3 by dialysis was found in normal subjects and in patients with untreated essential hypertension. The molecular weight of this inactive renin (43 000) is 2000 +/- 900 (S.E.M.) daltons higher (p less than 0.05) than that of naturally occurring active renin and the two forms of renin could be clearly separated by ion-exchange chromatography with diethylaminoethyl-"Sepharose'. Patients with low-renin essential hypertension had proportionately larger amounts of inactive renin than usual, so that the total renin in these patients approached the normal range. Inactive renin seems likely to be a form in which the enzyme is secreted since in a patient with renal-artery stenosis its concentration was greater in the renal-vein blood coming from the affected kidney.


Prevention of postoperative thromboembolism by dextran 40, low doses of heparin, or xantinol nicotinate. The efficacy of dextran-40 infusions or low-dose heparin or xantinol-nicotinate administration in preventing postoperative thromboembolic complications has been investigated in a prospective, controlled, randomised trial as part of an international multicentre trial. 382 patients over the age of forty years undergoing elective major general surgery were investigated; 100 had a complete protocol in the control, 94 in the heparin, 92 in the dextran, and 32 in the xantinol-nicotinate group. 31 patients died: 13 in the control group, 10 in the heparin group, 6 in the dextran group, and 2 in the xantinol-nicotinate group. In 4 controls, 6 in the heparin group, and 1 in the dextran group the cause of death was acute pulmonary embolism. 4 further controls, 1 in the heparin group, and 2 in the dextran group had emboli found at necropsy, which either contributed to the deaths or were incidental findings. The distribution of pulmonary emboli in this study did not show the difference between the control and the heparin group reported in the multicentre trial. The 125I-fibrinogen test was done in all patients. The frequency of isotopic deep-vein thrombosis was 36-0% in the controls, 12-8% in the heparin group, 21-7% in the dextran group, and 40-6% in the xantinol-nicotinate group. The difference between the controls and the heparin groups was highly significant and between the control and the dextran group, probably significant. There were significantly more side-effects in the heparin group than in the dextran group.


Bone-marrow transplant from an unrelated donor for chronic granulomatous disease. A boy with chronic granulomatous disease received a bone-marrow graft from a fully compatible unrelated donor, in association with a modified immunosuppressant regimen. He showed considerable clinical improvement with definite evidence of engraftment, and he has enjoyed a normal life for over 3 years.


Nephroptosis and hypertension.

Nephroptosis and hypertension. The degree of renal mobility was measured in patients with and without hypertension to evaluate a suggested causal relation between nephroptosis and hypertension. In contrast to men, women often showed pronounced renal mobility and the degree of renal mobility correlated positively with the blood-pressure. Fibromuscular dysplasia of the renal artery was always accompanied by a considerable degree of renal mobility on the side involved. The results of this study indicate that in women there may be an association between abnormal renal mobility and the development of high blood-pressure.


Upper gastrointestinal haemorrhage in hepatic cirrhosis: causes and relation to hepatic failure and stress. Emergency fibroscopy revealed bleeding lesions in 84 cirrhotic patients. In patients with moderate or no hepatic failure, the commonest actively bleeding sources were oesophagogastric varices and acute mucosal ulcers associated with the ingestion of anti-inflammatory drugs. In patients with severe hepatic failure, acute mucosal ulcers unrelated to drugs predominated and there was evidence that these were stress-induced erosions.


Effect of blood-group on relation between HLA match and outcome of cadaver kidney transplants. In a series of 4998 cadaver kidney transplants the extent of HLA matching correlated with transplant outcome in patients with blood-groups other than O (non-O) but not in patients of blood-group O. The high survival-rate of poorly matched kidneys in O recipients was responsible for the lack of correlation between HLA matching and graft survival in these patients. Survival-rates of transplants with 4 HLA mismatches were 52 +/- 3% at one year in 222 O-to-O grafts compared with 24 +/- 5% in 65 B-to-B grafts (P less than 0.0001). In 2827 non-O patients, there was a strong correlation between HLA matching and graft survival (P less than 0.0001 at one year). One possible reason for the cancelling out of the effect of HLA mismatching in type-O recipients is that these patients waited longer on dialysis for a transplant and consequently received more blood-transfusions.


Altered vitamin-D metabolism in pregnancy. Low levels of plasma-25-hydroxy-vitamin-D (25-OHD) (less than 16 nmol/1) were found in 33% of a group of pregnant women studied in South London. Non-White women in the last 10 weeks of pregnancy had significantly reduced levels compared with non-pregnant controls (P less than 0.02). Plasma-25-OHD was unrelated to dietary vitamin-D intake, age, parity, social class, plasma-calcium, and plasma-albumin. Reduction of plasma-25-OHD could contribute to the fall in plasma-calcium during pregnancy, and may result from enhanced maternal metabolism or increased utilisation of vitamin D by the fetus.


Mechanism of anti-inflammatory action of glucocorticosteroids. Glucocorticosteroids react with blood monocytes and tissue macrophages to produce a peptide factor which stimulates the random migration of polymorphs in vitro in the capillary-tube migration system. An identical effect on polymorph migration is produced by colchicine and vinblastine, drugs which inhibit the assembly of the cytoplasmic microtubules on which the functional activity of polymorphs depends. Pharmacological agents which inhibit microtubular assembly indirectly by increasing intracellular cyclic adenosine monophosphate (A.M.P.), also stimulate polymorph migration in vitro. These observations suggest that the anti-inflammatory activity of glucocorticosteroid drugs is mediated by a peptide hormone which inhibits polymorph microtubular assembly. Many peptide hormones are believed to act by increasing the concentration of cyclic A.M.P. within target cells and this mechanism is probably also responsible for the inhibitory effect of steroids on phagocytic cells.


Vaccination against whooping-cough. Efficacy versus risks. Calculations based on the mortality of whooping-cough before 1957 predict accurately the subsequent decline and the present low mortality. Notifications of incidence, though variable and incomplete, follow the same pattern of steady decline in the United Kingdom and are unaffected either by small-scale vaccination beginning about 1948 or by nationwide vaccination beginning in 1957. When valid comparisons can be made, attack-rates may be lower and complications fewer in vaccinated children, but allowance has to be made for overcrowding and socio-economic differences which may be more important as determinants of attack-rates. No protection by vaccination is demonstrable in infants. Adverse reactions and neurotoxicity following vaccinations were studied in 160 cases. In 79, the relationship to pertussis vaccine was strong. In 14 of these cases, reaction was transient but characteristic of a syndrome of shock and cerebral disturbance, which, in the other 65 cases, was followed by convulsions, hyperkinesis, and severe mental defect. It seems likely that most adverse reactions are unreported and that many are overlooked. Precise information about the efficacy and safety of this vaccine is lacking, because existing provisions, national and international, for epidemiological surveillance and evaluation are inadequate. The claim by official bodies that the risks of whooping-cough exceed those of vaccination is questionable, at least in the U.K.


Whooping-cough admissions to a paediatric hospital over ten years. The protective value of immunisation. 188 children with pertussis were admitted to Derbyshire Children's Hospital over a period of ten years. Fewer immunised children were admitted than would be expected if immunisation were ineffective. Immunisation seemed to decrease the risk of complications and the time spent in hospital. It is suggested that pertussis immunisation is valuable and should perhaps be introduced at an earlier age than is now recommended.