Ultrastructural cytochemistry of biogenic amines in nervous tissue: methodologic improvements. A new fixation method has been developed for localizing biogenic amines in nervous tissue. The method is a modification of the chromaffin reaction in which all fixation steps are buffered with mixtures of sodium chromate and potassium dichromate. In this way the fixation and cytochemical reaction are carried out almost simultaneously. Using the rat vas deferens as a model tissue, it was found that the preservation of electron dense (chromaffin) cores in the vesicles of adrenergic nerve terminals depended on several factors: a short primary fixation using low concentrations of aldehydes, the presence of the chromate/dichromate buffer during all fixation steps and, finally, a long incubation period in a slightly acidic (pH 6.0) solution of this buffer before postfixation in osmium tetroxide. Using this method it was possible to identify not only small and large dense-cored vesicles as storage sites for amines but also a tubular reticulum (neuronal endoplasmic reticulum), the latter especially in nerve terminals of mesenteric arteries and iris. Biogenic amines were also visualized in sympathetic ganglion cells and in the central nervous system e.g., supraependymal nerve terminals, tissues that up to now proved the most difficult in terms of amine localization. In all the tissues examined the cytochemical reaction was highly selective and present in well preserved tissue, which is a significant advance over previously available techniques. It therefore offers new opportunities for further studies on the role of biogenic amines as neurotransmitters.


A microspectrophotometric study of Papanicolaou-stained cervical cells as an aid in computerized image processing. As part of a study of cytologic automation, microspectrophotometric investigation of Papanicolaou-stained cervical cells was performed, using a Leitz MPV-II scanning photometer connected to a PDP 8/F minicomputer. It was shown that the selection of one single wavelength may result in difficulties in detecting boundries between background and cytoplasm and/or between cytoplasm nucleus. A set of two wavelengths, 530 nm and 570 nm, were found to be optimal for the image processing of these cells.


Quantitation of mast cell heparin by flow cytofluorometry. Mast cells can be automatically identified in a mixed cell population by flow cytofluorometry after Berberine sulphate staining. Volume specific counts of the total number of cells and number of mast cells, as well as frequency distributions of fluorescence intensities of mast cells, based on a large number of cells, can be rapidly obtained. Results obtained by microscope fluorometry of cells identified by phase contrast microscopy showviously published results it may be inferred that the fluorescence intensity of individual mast cells is proportional to mast cell heparin content. The automated cell counts correlated very well with manual hemocytometer counts. Both cell counts and the determination of mean mast cell fluorescence showed excellent reproducibility.


The identification of eosinophil colonies in soft-agar cultures by differential staining for peroxidase. There has been a need to easily quantitate the incidence of eosinophil colonies within soft agar cultures. This has been realized by layering of the agar with benzidine dihydrochloride that permits detection of peroxidase activity in cells. Eosinophil colonies can be specifically identified by the addition to the substrate of potassium cyanide, an inhibitor of enzyme activity in neutrophils and monocytes. The enumeration of eosinophil colonies can be accomplished by scanning fresh or embedded cultures with low power magnification.


The blue reaction product in horseradish peroxidase neurohistochemistry: incubation parameters and visibility. A blue reaction product is formed at sites that contain horseradish peroxidase (HRP) activity when benzidene is used as the chromogen. With neutral red as a counter stain, this method affords excellent visualization of both retrograde and orthograde axonal transport of intracerebrally injected HRP. The visibility of this blue reaction-product is better than the visibility of the brown reaction-product obtained in the commonly used diaminobenzidene procedures. Variations in incubation times and reagent concentrations resulted in significant differences in the extent to which transported HRP could be demonstrated with benzidene. One of these benzidene procedures demonstrated a wider extent of HRP transport than a representative diaminobenzidene procedure. The substantia nigra and the nucleus locus ceruleus did not display artifactual deposition of the blue reaction-product.


Mapping of the genetic control of murine response to low doses of the dinitrophenyl conjugates of ovomucoid and bovine gamma-globulin. The antibody responses in mice to low doses of the DNP-conjugates of ovomucoid (OM) and bovine gamma-globulin (BGG) were measured for a number of inbred strains carrying independent and recombinant H-2 haplotypes. This permitted mapping the gene(s) controlling the response to low doses of OM (Ir-1-OM) within the I-A subregion of the H-2 complex. Similarly, it was possible to map the primary gene(s) controlling the response to low doses of BGG (Ir-1-BGG) within the I-A and/or I-B subregions. Further localization of the Ir-1-BGG gene(s) to the I-A or I-B subregion of the H-2 complex was not possible due to the ambiguous response of the B10.A(4R) strain of mice.


Relationship between beta2-microglobulin and cell-surface alloantigens of the mouse. Molecular relationships between beta2m and other cell surface antigens (H-2, Tla, Ia, and Thy-1) were studied with the double immunofluorescence method. Cells were incubated with an antiserum against one antigen capped, and then tested with an antiserum against a second antigen. Capping of beta2m on thymocytes led to simultaneous capping of H-2 and Tla but not Thy-1 antigens; capping of H-2 and TIa (but not Thy-1) antigens resulted in capping of all beta2m detectable by the immunofluorescence method. Similarly, capping of beta2m on B or T lymphocytes resulted in capping of H-2 and vice versa. Ia antigens on B lymphocytes were not capped after the redistribution of beta2m. We conclude from these data that, in the cell membrane of thymocytes, virtually all the beta2m molecules are associated with H-2 and Tla, but not with Thy-1, and that on the cell surface of T or B lymphocytes, virtually all beta2m  is associated with H-2 but not with Ia. We found no evidence of any significant free beta2m on either thymocytes or splenocytes.


Development of mast cells in vitro. II. Biologic function of cultured mast cells. Mast cells were obtained by long term culture of rat thymus cells on rat embryonic fibroblast monolayers. Pure mast cell preparations obtained culture were incubated with 125I-labeled rat E myeloma protein to study receptors for IgE on their surface. When the cells were obtained after 35 to 45 days culture, the average number of receptors per mast cell was 100,000 to 400,000. An equilibrium constant of the binding reaction between their receptor and rat IgE was in the order of 108 M-1. The histamine content of the cultured mast cells was 0.2 to 5 mug/106 cells. The measurement of histamine content in mast cells recovered after different periods of culture suggested that the histamine content increased with maturation. Even after 45 to 50 days culture, the histamine content of cultured mast cells was significantly lower than that in rat peritoneal mast cells. The cultured mast cells were passively sensitized in vitro with rat IgE antibody against Nippostrongylus brasiliensis. The sensitized cells released histamine upon incubation with the antigen. It was also found that cultured mast cells released histamine upon exposure to compound 48/80. These results indicated that cultured mast cells have physiologic functions similar to those of normal rat mast cells, but they have not reached full maturation.


Conformational integrity of myoglobin after immunization with Freund's adjuvant. The use of Freund's complete adjuvant for immunizing goats with myoglobin produces mainly antibodies directed against antigenic determinants present in the native protein. Only about 9% of the total antibodies produced are directed toward determinants not expressed in tha native molecule. This shows that neither emulsification nor the subsequent in vivo events leading up to the immune response appreciably perturb the conformation of the protein surface.


Amyloid-related serum protein SAA from three animal species: comparison with human SAA. The amyloid-relates serum protein SAA has been isolated by gel filtration in 10% formic acid from three animal species: mink, mouse, rabbit. Sera used in the isolation procedure were obtained from animals in which high concentrations of SAA had been induced by treatment with LPS. The isolated SAA proteins had a subunit size similar to that of human SAA, with m.w. values ranging from 10,000 to 11,700 (estimated by gel filtration in 6 M guanidine-HC1) or 12,400 to 15,000 (estimated by SDS-PAGE). The m.w. studies and amino acid sequence data indicated that SAA and the amyloid fibril protein AA in the mouse, and probably also the mink, are related in the same way as in man, the two proteins having common NH2-terminal amino acid sequences and SAA being extended by 20 to 40 residues at the COOH-terminal end of the molecule.


The immunologic and molecular profiles of HLA antigens isolated from urine. Human urine was shown to be a good source for the isolation of immunologically functional HLA-A9 antigens. The use of complex solubilization procedures can be avoided since the antigens are present in soluble form and are not complexes with membrane fragements. Purification in excess of 400-fold could be achieved by the application of cellulose ion exchange chromatography, isoelectric focusing, and acrylamide gel electrophoresis. The purified HLA-A9 antigen is composed of a glycoprotein of m.w. 38,000 and beta2-microglobulin, a peptide of m.w. 12,000. HLA-A9 antigens isolated from urine proved to be immunologically functional since they not only reacted specifically with anti-HLA-A9 alloantibody but also elicited anti-HLA-A9 xenoantibodies. These antibodies when covalently attached to Sepharose 4B specifically bound HLA-A9 antigens isolated from both serum and urine.


Isolation and characterization of canine secretory immunoglobulin M. Canine secretory immunoglobulin M, isolated from both colostrum and bronchial secretions, contained the unique glycoprotein bound secretory component. The presence of this extra subunit accounted for the differences in size, quaternary structure, and antigenicity observed upon comparison of secretory immunoglobulin M with its serum counterpart. Approximately 90% of the isolated secretory immunoglobulin M contained covalently bound secretory component while, in the remainder of the population, secretory component was loosely attached and easily dissociated from the immunoglobulin. Following peptide bond cleavage with cyanogen bromide, the release of bound secretory component and J chain from secretory immunoglobulin M was not detected. Because cyanogen bromide cleavage of secretory immunoglobulin A results in the release of these subunits, differences in the primary structure of secretory immunoglobulin M and secretory immunoglobulin A must exist around the binding sites for secretory component and J chain.


Coincident hepatitis B surface antigen and antibodies of different subtypes in human serum. Three patients with simultaneously detectable hepatitis B surface antigen (HBsAg) and antibody (anti-HBs) in their sera were studied for subtypes of HBsAg and anti-HBs. In each case anti-HBs was found to be directed to a different subtype than that of the circulating HBsAg, indicating that reinfection (or simultaneous infection) with a second subtype occurred.


Affinity in radioimmunoassay of antibody cross-reactive with carcinoembryonic antigen (CEA) and colon carcinoma antigen-III (CCA-III). Several antisera raised against purified carcinoembryonic antigen (CEA) were evaluated for their content of antibody cross-reactive with colon carcinoma antigen-III (CCA-III). All antisera gave a reaction of partial identity between CEA and CCA-III and demonstrated a high titer in CEA radioimmunoassay (RIA). Between 50 and 70% of the CEA RIA activity was removed, however, by absorption with soluble CCA-III or adsorption onto CCA-III-containing immunoadsorbents. Immunoadsorbent retained antibody gave a line of complete identity between CEA and CCA-III. Purified CCA-III (2 mug) only partially depressed CEA binding by this common site antibody, whereas nanogram quantities of CCA-III inhibited the reaction between specific CCA-III antibody and radioiodinated CCA-III. In addition, low levels of CEA were equally effective in depressing CEA binding by the common site or CEA-specific antibody. The higher affinity in RIA of the common site antibody for CEA over CCA-III suggests that the common determinant expressed on CEA is stereochemically different from that on CCA-III. The results further demonstrate that interference by plasma CCA-III is not a significant factor in the measurement of CEA by RIA.


A micro-titer red-cell-linked-antigen-antiglobulin-test for DNP specific immunoglobulins. A micro-titer red-cell-linked-antigen--antiglobulin test is described for the measurement of DNP specific immunoglobulins in mouse plasma samples.


Immunofluorescent localization of basement membrane in lesions of dermatitis herpetiformis. Dermatitis herpetiformis (DH) is a blistering disease with a characteristic histology that includes papillary edema, neutrophilic papillary microabscesses, and development of subepidermal blisters. In spite of this pathologic sequence occurring entirely beneath the basement membrane zone, prior studies have indicated that the basement membrane, as defined by period acid-Schiff (PAS) or silver stains, lies at the floor of fully formed blisters or is destroyed by the disease process. To more accurately assess its location in primary lesions of DH, the basement membrane was stained using immunofluorescent techniques. Lesional skin from 5 patients with DH was used as substrate for indirect immunofluorescence with sera from patients with bullous pemphigoid (BP) and fluoresceinated antihuman IgG. The BP-stained basement membrane was attached to the roofs of early blisters, where it would be expected from the pathologic sequence of blister formation. PAS stains of the same or serial sections show the basement membrane to be in the roof or at the floor of the blisters. PAS stains of sections from formalin-fixed lesional skin, on the other hand, show the basement membrane to routinely lie at the blister floor, when not destroyed. The BP-stained epidermal basement membrane has greater anatomic and functional significance than either the PAS-or silver-stained basement membrane for two reasons: (1) it corresponds to a specific morphologic structure, the lamina lucida, a part of the epidermis, and remains attached to the rest of the epidermis unless destroyed; and (2) it is antigenic, capable of binding with BP antibodies.


Quantitative analysis of the major subdeterminants of hepatitis B surface antigen. Antisera monospecific for subtyping of speciments of hepatitis B surface antigen (HBS Ag) were prepared by affinity chromatography. These sera were then used for quantitation of the a, d, and y subdeterminants of HBS Ag on a standard panel of sera and on 41 unselected sera containing HBS Ag. Although 25 of this latter group were completely monospecific for ad or ay, the remaining 16 sera showed degrees of cross-reaction with the heterologous antiserum. It is suggested that d and y subdeterminants are not infrequently present on the same particle.


A passive hemagglutination test for diagnosis of trench fever due to Rochalimaea quintana. A passive hemagglutination test devised for diagnosis of trench fever was easily performed and highly sensitive and specific. Tanned sheep erythrocytes were sensitized with soluble antigen from Rochalimaea quintana. The test detected antibody in six of seven cases of primary infection and in four cases of late, relapsed trench fever. Titers of antibody ranged from 1:20 to 1:640. Although both IgM and IgG antibody to R. quintana were detected by passive hemagglutination, IgG appeared to be the major reactive antibody. Antigens involved in the reaction were two types of proteins, one inactivated at 50 C and 60 C and the other at 80 C and 100 C. Of 322 control samples of sera that were tested, only one reacted positively; thus, the test had a specificity of greater than 99%. The single positive reaction was in serum from a patient with Q fever. This finding suggests that, in an area where Q fever is endemic, this disease must be ruled out in the interpretation of a positive passive hemagglutination test. Sera should be tested routinely against tanned, unsensitized erythrocytes, since an occassional sample of serum may agglutinate unsensitized cells. Because of its sensitivity and specificity, as well as its simplicity of performance, the passive hemagglutination test shows promise as a useful procedure for serologic identification of both acute and past infection with R. quintana.


Histocompatibility antigens in patients with leprosy. The frequencies of distribution of 25 histocompatibility antigens were determined in 92 Mexican patients with leprosy and compared with those in 315 Mexicans who did not have the disease. No statistically significant differences were found between the patients and the controls in regard to histocompatibility antigens, and subgroups with a significant difference could not be identified by division of the patients according to the density of Mycobacterium leprae or the presence or absence of cell-mediated immunity directed against antigens of M. leprae.


Radiation leukemia in C57BL/6 mice. I. Lack of serological evidence for the role of endogenous ecotropic viruses in pathogenesis. The humoral immune response against endogenous ecotropic murine leukemia viruses (MuLV) was examined in irradiated and control C57BL/6 mice. Control mice developed antibodies against MuLV slowly throughout life. In contrast, within 2-3 mo after irradiation 90% of irradiated C57BL/6 mice had developed detectable antibodies against MuLV. The characteristics of this immune response, however, were identical in control and irradiated mice in terms of peak titers, specificity for endogenous ecotropic MuLV, and reactivity against the ecotropic viruses' glycoprotein (gp71). Moreover, the rate of appearance of antibodies against MuLV in irradiated mice and the peak titers were generally not affected by age at irradiation, dose of irradiation (two, three, or four treatments of 175 R), or bone marrow reconstitution. Although the ability of irradiation to accelerate the appearance of antibody in a population of C57BL/6 mice suggested activation of endogenous ecotropic MuLV, there was no apparent correlation between the appearance of this immune response or its persistence and the development of lymphoma. Thus, the incidence of lymphoma was comparable in mice that: (a) developed no immune response; (b) developed an immune response only transiently after irradiation; or (c) developed an immune response which persisted until death from lymphoma. Moreover, experimental conditions that alter the ability of irradiation to induce leukemia, such as age, dose, or bone marrow reconstitution did so without significantly altering either the rate of appearance of a humoral immune response to MuLV or its peak titers. The results, therefore, fail to demonstrate any seroepidemological relationship between endogenous ecotropic MuLV and radiation-induced leukemia.


Radiation leukemia in C57BL/6 mice. II. Lack of ecotropic virus expression in the majority of lymphomas. The expression of endogenous ecotropic viruses in radiation-induced thymomas of C57BL/6 mice was examined. Competition radioimmunoassays for AKR MuLV gp71, p30, and p12 were used for viral antigen expression. 3 of 40 lymphomas had readily detectable ecotropic gp71 at levels of 95-689 ng/mg protein; the remainder of the tumors had no detectable gp71 (less than 1.0 ng/mg protein). 30 thymomas were characterized by the presence of MuLV p30 at levels of 1-10 ng/mg protein, levels that were comparable to those found in thymus extracts from age-matched, nonirradiated control. 10 tumors were characterized by having p30 levels of 10-30 ng/mg protein. In one tumor significant levels of AKR MuLV p12 were detectable. Since B-tropic and N-tropic viruses from C57BL/6 mice have glycoproteins (gp71) indistinguishable from AKR MuLV gp71 and the N-tropic virus had a p12 serologically identical to AKR MuLV p12, these results demonstrate that overt endogenous B-tropic virus was detectable in 2 of 40 thymomas and endogenous N-tropic virus was detectable in 1 of 40 thymomas. The lack of overt expression of gp71 or p12 was also confirmed by cytotoxicity assays using monospecific antisera to these viral proteins. Radiation-induced lymphomas were also examined for the presence of reverse transcriptase after chromatography of tissue extracts on poly G-Sepharose. One tumor, which was characterized by the lack of gp71, also had no detectable reverse transcriptase; whereas one tumor with gp71 was characterized by readily detectable levels of reverse transcriptase in cellular extracts. The presence of viral RNA was examined using AKR cDNA. Low levels of RNA capable of hybridizing with AKR cDNA were found in age-matched, nonirradiated mice; these hybrids had Tm's of 72 degrees C, while hybrids with AKR MuLV 70S RNA had Tm's of 80 degrees C. In 1 of 12 thymomas the concentration of hybridizable RNA and the Tm of the hybrids were identical to control values. In 9 of 12 thymomas the concentration of hybridizable sequences increased approximately three-to fivefold and the Tm of these hybrids varied from 73 to 75 degrees C. In 1 of 12 thymomas the concentration of hybridizable sequences increased over 100-fold, hybridized completely with AKR MuLV cDNA, and the hybrids had Tm's of 79 degrees C. This thymoma was also characterized by the presence of the AKR MuLV type of gp71 and p12. One tumor was characterized by a 10-to 100-fold increase in hybridizable sequences, which only partially hybridized with AKR MuLV cDNA, and hybrids had a Tm of 73 degrees C. This tumor was characterized by the presence of AKR MuLV gp71 but not AKR MuLV p12. The results taken together demonstrate that overt endogenous ecotropic virus expression is only rarely detectable in radiation-induced thymomas of C57BL/6 mice.


The specificity of cellular immune responses II. The structure of antigenic determinants leading to T-lymphocyte stimulation. T cells from guinea pigs immunized with the hapten 2,4-dinitrophenyl (DNP)-coupled directly to mycobacteria are of interest since they recognize and respond to DNP conjugated to many but not all carriers. The experiments reported here further analyze the structure of the complex, chemically defined antigenic determinants recognized by such T cells. These antigenic determinants can have DNP coupled either to the xi-amino group of lysyl residues or to the hydroxyl group of tyrosyl residues. Furthermore, essential contributions to the determinant recognized by such T cells are made by amino acid residues to which the hapten is not attached. Such residues are thought to be close to the hapten group itself, since introducing a small spacer between hapten and carrier prevents recognition. The hapten itself is also recognized and discriminated from other haptens with great precision by these T lymphocytes. The strain of guinea pig immunized affects the precise specificity characteristics of the responding T cells, in a way that may reflect the activity of histocompatibility-linked immune response genes. Finally, the characteristics of the immunogen have been studied. It is thought that the lipid content of the mycobacteria may be critical in inducing the hapten-reactive T cells, and this is supported by finding similar responses in T cells from guinea pigs immunized with DNP protein to which lipid has been covalently attached. Thus, the T-cell population being studied, while recognizing haptens with great precision, appears to require a larger determinant for activation than do hapten-specific B lymphocytes.


The specificity of cellular immune responses in guinea pigs. III. The precision of antigen recognition by T lymphocytes. T lymphocytes from guinea pigs immunized with 2,4-dinitrophenyl (DNP) derivatives of mycobacteria respond to a variety of DNP conjugates. Preincubation of such cells with a given DNP conjugate under conditions which lead to the inactivation of responding cells causes a loss of the response to that conjugate, but has little effect on the response to DNP coupled to unrelated carriers. Thus, the responses of such cells to a variety of DNP conjugates can best be explained by the presence of a mixture of highly specific cells each responding to a different antigenic dterminant rather than by the presence of T cells with specificity limited to the hapten itself. Furthermore, the activity of T cells from DNP-mycobacteria-primed donors could not be blocked by a variety of nonstimulatory DNP conjugates. This suggests that while such T cells clearly recognize DNP with great precision, the receptor does not contain a very high affinity site for the hapten. A possible model for such a T-cell receptor is discussed.


Some properties of antisera to serum amyloid A protein (SAA): inhibition of precipitation by complexing of SAA to albumin. Three potent rabbit antisera to human serum amyloid A protein (SAA) appear to be directed exclusibely to the carboxy terminal region not shared with the tissue amyloid A protein. Since binding to albumin completely blocks the reaction of these antisera with the antigen, and since SAA exists in serum complexed to albumin, the anti-SAA cannot be used to detect or quantitate this serum component. The possibility that similar problems will be encountered with immunoassays for molecules that exist complexed to other proteins is discussed.


Genetic control of the immune response to staphylococcal nuclease. IV. H-2-linked control of the relative proportions of antibodies produced to different determinants of native nuclease. The relative proportions of antibodies of different specificities within antisera raised to native staphylococcal nuclease have been studied in several strains of mice in which the antibody response has been shown to be under H-2-linked Ir-gene control. A method was developed in which binding to different radiolabeled fragments of nuclease was titrated against increasing fragment concentration until the binding capacity of the antiserum for that fragment was saturated. In comparing the low responder (H-2b) strain C57BL/10 with its congenic high responder counterpart B10.A (H-2a), it was found that the two strains made markedly and reproducibly different proportions of antibodies to different determinants on native nuclease. Since these two strains differ only at H-2, and therefore have identical immunoglobulin structural gene repertoires, we conclude that H-2-linked Ir genes can control the response to different determinants on the same antigen molecule independently of one another. This result suggests a possible role of H-2-linked genes in the selection of specific B cells.


The Fc receptor on thymus-derived lymphocytes. IV. Inhibition of binding of antigen-antibody complexes to Fc receptor-positive T cells by anti-Ia sera. Treatment of splenic T lymphocytes with anti-Ia antiserum inhibits the binding of antigen-antibody (AgAb) complexes to the majority (less than 50%) of Fc receptor-positive (FcR+) T cells. A similar inhibition was observed with anti-H-2D and anti-H-2K sera but not with anti-Thy 1.2. Despite the presence of Ia determinants on peripheral T cells, as established by the inhibition of AgAb binding, Ia could not be detected on peripheral T cells by immunofluorescence assays. Data obtained with the AgAb-binding inhibition assay indicate that determinants controlled by loci mapping in the I-A and I-C, S, or G regions are present on the FcR+ T cells. Evidence is presented that subpopulations of T cells within the FcR+ T-cell population may be distinguishable on the basis of which I-region-controlled determinant is expressed. The data are discussed in terms of phenotypic and functional heterogeneity of T lymphocytes.


Qualitative patterns of protein synthesis in the mouse oocyte. Mouse oocytes were found to synthesize proteins actively at the germinal vesicle, metaphase I, metaphase II, and pronuclear (6 hours post-fertilization) stages. The qualitative pattern components being synthesized in vitro, as demonstrated using polyacrylamide gel electrophoresis, changed throughout maturation and fertilization. Oocytes were arrested at metaphase I by greater than 0.1 mug/ml cycloheximide or actinomycin D. The protein pattern in oocytes cultured in the presence of actinomycin D progresses to a metaphase II pattern in spite of the nuclear maturation arrest, indicating a dissociation between meiotic maturation and the changes in the pattern of proteins synthesized at different stages of maturation.


A comparison of four methods used to concentrate Rous sarcoma virus from tissue culture fluids. Three methods of pelleting, pelleting followed by Pronase treatment, polyethylene glycol (PEG)-Pronase, and diaflo ultrafiltration (diafiltration) were used to concentrate RSV(RAV-1) from tissue culture fluids. Sucrose-gradient fractions containing virus preparations which had been concentrated by diafiltration or pelleting were heavily contaminated with amorphous debris. This debris was not present in similar, gradient-purified preparations that had been concentrated by the PEG-Pronase or pellet-Pronase methods. Maximum recovery of radiolabelled virus particles and virion-associated RNA-dependent DNA polymerase activity was obtained in gradient fractions containing virus concentrates prepared by the pellet-Pronase and PEG-Pronase methods. Although there were slight differences in recovery by these two methods, the advantages of the PEG-Pronase method make it the preferred method, especially when large volumes of tissue culture fluids are used.


Effects of a memory aid of the three types of conservation in institutionalized retarded children. Qualitative identity, quantitative identity, and equivalence conservation were assessed in 60 retarded boys and girls in three groups (MAS 5.4,6.3, and 7.5)9 Half of the Ss were provided a memory aid while the other half were not. The memory aid did not facilitate conservation on any of the tasks. The order of task difficulty found was qualitative identity, quantitative identity, and equivalence conservation. These data were discussed in terms of memory deficits and factors contributing to conservation and M.A.


Neuron culture from adult goldfish.

Neuron culture from adult goldfish. Neurons and glia from the central nervous system of the adult teleost Carassius auratus have been grown as explant cultures of minced brain tissue and as trypsin dissociated cells. These cultures exhibit extensive neurite growth from two neuronal types, have organotypic ultrastructure, and contain electrically active cells. Autoradiographic data indicate that these neurons do not divide in culture, and histological evidence suggests that some mature neurons survive explantation and regenerate processes. However, explantation of brain fragments not containing undifferentiated cells, localized in the ventricular and subventricular zones in the brains of fish, resulted in mesenchymal and glial cell cultures only. Therefore, a contribution to the population of cells in culture by undifferentiated cells must be considered. The cultured neurons remained viable for at least 19 weeks and ultrastructural and electrophysiological data indicate synaptic interaction between cells in explant cultures.


Isoelectric focusing and electrophoresis of the CSF proteins in tremor of different origins. The CSF proteins have previously been very little investigated in the cerebellar syndrome of chronic alcoholism and in essential tremor. Such studies have been carried out more thoroughly by electrophoretic methods in Parkinson's disease but generally with normal results. In the present investigation the CSF proteins were examined by isoelectric focusing and quantitative paper electrophoresis in 10 patients with the cerebellar syndrome of chronic alcoholsm, 12 patients with Parkinson's disease and 16 subjects with essential tremor. Abnormal CSF proteins of very similar appearance were found on isoelectric focusing in the acidic pH interval 5.6-5.8 in 80% of the patients with the cerebellar syndrome of chronic alcoholism. In Parkinson's disease the most common aberration was evidence of nonspecific blood-CSF-barrier damage which occurred in half of the patients. In only 17% of these cases did other alterations appear, situated in the pH range alkaline to pH 5.8. Abnormal CSF proteins were found in 94% of the patients with essential tremor. The aberrant proteins appeared in both the acidic and alkaline pH regions, most frequently with anisoelectric point at pH 5.9, 7.2 and 9.3. There was a considerably higher frequency of CSF protein abnormalities in different pH ranges in patients with tremor of more pronounced degree as compared to those with only mild symptoms. The electrophoretic examinations failed to show any conclusive alterations. Barrier-damage patterns of mild or moderate degree or slightly increased levels of CSF beta1-globulin were occasionally found in all 3 diseases. The results indicate that isoelectric focusing of the CSF proteins may be of diagnostic value in the cerebellar syndrome of chronic alcoholism and in essential tremor but does not reveal any characteristic abnormalities in Parkinson's disease.


Steroidal compounds (injectable and implants) affecting spermatogenesis in men. The effect of androgen and different progestins on spermatogenesis was studied in young men with subfertility, prostatovesiculitis and haematospermia, and in older men with benign hypertrophy of the prostate. The compounds (testosterone, testosterone oenanthate, ethinyl oestradiol, megestrol acetate, ethinyl norgestrienone and Depo-Provera) were administered intramuscularly, orally or as subcutaneous silastic implants.


Protein-binding polyhedral boranes.

Protein-binding polyhedral boranes. A series of polyhedral borane derivatives containing protein-binding functional groups has been synthesized. Problems encountered in earlier studies (low incorporation levels, gross precipitation of conjugates) have been overcome by including a water-solubilizing gluconamide group in the structure. This modification has allowed high levels of boron to be covalently bound to HGG, forming a completely water-soluble conjugate.


Antenatal diagnosis of trisomy 13 with unexpected increase in alpha-feto protein. A 24-year-old woman who had previously given birth to an infant with Down's syndrome was shown by chromosomal analysis of the liquor amnii to be carrying an infant with trisomy D. Routine examination of serum and liquor alpha-feto protein (AFP) in the antenatal period showed unexpected high levels of both, consistent with a neural tube defect. The fetus, however, did not have evidence of a neural tube defect but had scalp defects which were presumed to have allowed the leakage of AFP from the fetus into the liquor amnii and hance into the maternal serum.


A serotyping system for Clostridium welchii (C. perfringens) type A, and studies on the type-specific antigens. A serotyping scheme for Clostridium welchii (C. perfringens) type A employing 57 antisera has been used to investigate the epidemiology of 153 food-poisoning outbreaks and 32 cases of gas gangrene and other clinical infections. Respectively 65% and 59% of the isolates were typable, and in 55% of the food-poisoning outbreaks the causative serotypes were established. Isolation and reporting methods that would render the typing scheme of even greater epidemiological value are described. The type-specific antigen was shown to reside in the capsule and to be lost from strains that had become rough. Development of roughness and its prevention are described. A great range of antisera and an internationally acceptable serotyping scheme is expected after integration of this set with those developed independently in America and Japan.


Xenotropic properties of an isolate from murine Rauscher leukemia virus in primates. Murine Rauscher leukemia virus (MuRLV) from BALB/c plasma consisting of a mixture of an ecotropic and a xenotropic virus could be separated out by a selection process when propagated in human and simian cell cultures. This hypothesis is supported by obtaining consistently lower infectivity titers of human cell propagated RLV in human and simian cells as compared to MuRLV propagated in mouse cell cultures. Furthermore, RLV passaged in a simian cell culture failed to replicate in mouse cells, had a wide host range, was able to rescue Moloney sarcoma genome, possessed murine type C group-specific antigen, and was neutralized by anti-HRLV. Its reverse transcriptase was strongly inhibited by antiserum to MuRLV enzyme; however, antiserum to woolly monkey enzyme also inhibited (30%) its reverse transcriptase, suggesting some difference in antigenic properties. Inoculation of this virus in rhesus monkeys was inconclusive.


Carcinogenicity of aflatoxin B1 in rhesus monkeys: two additional cases of primary liver cancer. Three of 42 (7%) monkeys given aflatoxin B1 (AFB1) for longer than 2 years have developed primary malignant neoplasms of the liver. Liver biopsies performed at intervals during aflatoxin administration revealed that neoplasia was preceded by pathologic lesions of the liver, including toxic hepatitis, proliferation of pseudotubules, and hyperplastic nodules. Serum alpha-fetoprotein levels, monitored in one of the monkeys by radioimmunoassay, paralleled tumor growth and recurrence of the hepatocellular carcinoma. Normal serum alpha-fetoprotein levels were noted for a monkey with hemangioendothelial sarcoma. Our results implicate AFB1 as a liver carcinogen in monkeys and add additional support to the hypothesis that humans exposed to this substance may be at risk of developing liver cancer.


Interaction analysis of selective and nonselective cell-mediated cytotoxicity. The coexistence of selective and nonselective cytotoxic cells in effector suspensions required a method of separating the two effects before specificities in cell-mediated cytotoxicity could be investigated. A method of analysis was derived which used the average cytotoxicity for each effector and target to estimate selective and nonselective cytotoxic effects. The analysis clearly detected specificity in tests of cell-mediated lympholysis, and application to tests of cell-mediated cytotoxicity on cultured human tumor cells showed that selective reactions were found.


Host response to tumor-associated fetal antigens: kinetics and components. Using a colony-inhibition bioassay that measures the contribution of the host response in transplantation immunity to tumor-associated fetal antigens (TAFA), I evaluated the occurrence of TAFA during different developmental stages of murine fetal liver cells. A maximum concentration of TAFA was present at approximately 15 days' gestation. A similar age-equivalence phase of gestation was also found in pig fetal liver. Host-response kinetics to TAFA were examined, and results indicated a similar pattern of activity for tumor- or fetal-immunized groups with a transient immune suppression in the middle of the study period. I detected specific antibodies generated by tumor and fetal cell immunization by a complement-dependent serum microcytotoxicity assay. The specificity of the reaction was evaluated further by absorption studies that demonstrated specific cross-reactive antibodies to fetal and tumor cell targets. Adoptive transplanted immunity to TAFA was indirectly evaluated by a modified Winn assay which indicated that a cellular immune mechanism was an important component of the host response to TAFA.


In vitro induction of tumor-specific immunity. II. Activation of cytotoxic lymphocytes to murine oncofetal antigens. The presence of oncofetal antigens (OFA) on a wide variety of murine tumor cells was demonstrated to a totally in vitro system of cellular immunity. Nonimmune spleen lymphocytes were cocultivated with irradiated syngeneic fetal liver cells and, at various times after initiation of culture, were tested for the presence of cytotoxic lymphocytes (CL) by 51Cr-release assay with labeled tumor target cells. Significant cytotoxic activity was regularly detected after such culture, whereas only minor levels appeared in control cultures of spleen lymphocytes with irradiated syngeneic spleen cells. Specificity of the reaction was assessed by inhibition tests in which nonlabeled cells were admixed to the CL and 51Cr-labeled tumor targets. Fetal liver cells gave significant inhibition; however, no inhibition was found with adult spleen cells. Various tumor types gave inhibition, and fibrosarcomas were more effective than plasmacytomas or lymphomas. The results suggested that all tumor types tested possess such OFA, as well as their unique or virus-associated, tumor-associated transplantation antigens, and that the in vitro system permits a more active response to the tumor-associated OFA than that observed in in vivo studies.


Induction of tumors in Syrian hamsters by a human renal papovavirus, RF strain. Injection of RF virus (RFV), a papovavirus isolated from human urine, into newborn Syrian hamsters induced subcutaneous sarcomas in 50% of the recipients with 18- to 48-week latent periods. Transplantation of 2 X 10(6) primary RFV-induced tumor cells into weaning hamsters caused tumors in 100% of the recipients within 1-2 weeks. Continuous tissue culture cell lines were established from two primary tumors; one of these was transplantable. An in vitro-transformed continuous cell line (RF-194) obtained by infection of primary hamster embryo fibroblasts with RFV was transplantable in weaning hamsters. Neither infectious RFV nor virion antigens were detected in transformed cells. No RFV was recovered when transformed cells were fused with permissive, human embryo kidney cells by means of inactivated Sendai virus. Immunoperoxidase staining was used to show that all three RFV-transformed cell lines contained an intranuclear T-antigen closely similar to that of simian virus 40(SV40)-infected cells. Most hamsters (84%) with primary or transplanted RFV tumors responded with antibodies that reacted with RFV T-antigen and the T-antigen of SV40-infected cells. Likewise, hamster antisera against SV40 T-antigen cross-reacted with RFV T-antigen. Adsorption of RFV T-antisera with an excess of lyophilized SV40-transformed cells removed all detectable activity against SV40 T-antigen but left significant activity against RFV T-antigen. The reciprocal adsorption produced an antiserum spedicic for SV40 T-antigen. Thus human and simian papovavirus T-antigens were related but immunologically separable.


Effect of ethidium bromide on transplanted virus-induced tumor cells. Ethidium bromide (2,3-diamino-5-ethyl-6-phenylphenanthridinium bromide) significantly inhibited the RNA-dependent DNA polymerase of types A and C particles isolated from transplanted adenovirus 12-induced tumors of CBA mice. It was also cytotoxic for an established in vitro line of adenovirus 12-induced tumor cells of CBA mice and caused cell death, inhibition of [3H]thymidine uptake, and a significant reduction of cells in metaphase. Ethidium bromide significantly inhibited the in vivo growth of transplanted adenovirus 12-induced tumor cells of CBA mice, simian virus 40-induced tumor cells of hamsters, and murine leukemia virus-induced lymphoma cells of BALB/c mice. The compound may have exerted the antitumor activity by selectively affecting oncornavirus in the tumor cells.


Detection, quantitation, and characterization of the major internal virion antigen of the bovine leukemia virus by radioimmunoassay. The major internal polypeptide of the bovine leukemia virus (BLV) was purified to homogeneity with the use of gel filtration and affinity chromatography. Like previous results, the protein had a molecular weight of 25,000 daltons as determined by electrophoresis in polyacrylamide gels with sodium dodecyl sulfate. More than 90% of the 125I-labeled protein was precipitated by bovine sera that reacted in immunofluorescence tests with acetone-fixed BLV-infected cells. In contrast, minimal precipitation (less than 5%) was observed with sera from 36 cattle in leukemia-free herds; these sera, negative by immunofluorescence, included six samples that had high titers of antibodies to the foamy-like bovine syncytia virus (BSV). Antisera prepared against several other oncornaviruses or the Mason-Pfizer monkey virus (M-PMV) did not bind the BLV p25 protein. Conversely, the labeled p30 polypeptides of several oncornaviruses tested did not react with bovine sera that had high titers of antibodies to BLV p25. Competitive radioimmunoassay(s) (RIA) also failed to detect cross-reactions between BLV p25 protein and the internal polypeptides of other mammalian and avian oncornaviruses, M-PMV, or foamy-like BSV. The RIA for BLV p25 antigen was also highly sensitive and specific for the detection and quantitation of the antigen in virus preparations and cell homogenates.


Safety of viral vaccine cell substrates: a reevaluation. Primary cultures of African green monkey kidney and rabbit kidney as well as diploid cell lines WI-38 and DBS-FRhL-2 were examined for evidence of tumorigenicity and latent RNA tumor viruses. Cells inoculated into immunosuppressed newborn hamsters and rhesus monkeys were not tumorigenic. Cells treated with 2'-deoxy-5-iodouridine to induce the production of latent viruses were examined by electron microscopy, density gradient centrifugation, and the reverse transcriptase enzyme assay. No evidence was found for RNA tumor viruses by the biochemical or biophysical methods used. The results indicated that each type of mammalian cell currently used in the production of virus vaccines would be acceptable for these parameters of safety if similar control procedures were applied at the time the vaccines were manufactured.


Potentiation of cytotoxic T-cell function by virus. Inoculation of C57BL/6J mice with allogeneic P815 mastocytoma cells in the presence of simian virus 40 (SV40), a DNA tumor virus, led to an enhanced cytolytic T-cell response to P815 in vivo. Cytotoxic function was also augmented if SV40 was given subsequent to a primary immunization, even when mice were given a suboptimal dose of immunizing cells. Although SV40 increased the cell-mediated immune response to allogeneic cells, it did not enhance the antibody response to the soluble antigen dinitrophenyl bovine gamma-globulin, a helper T-cell-dependent response. Thus it appeared that SV40 had a selective adjuvant effect on lymphocyte subpopulations, since it increased cytotoxicity but not helper T-cell function.


Role of divalent ion complex formation in pyran--inhibition of nucleic acid biosynthesis. The degree of inhibition of mammalian DNA-dependent RNA polymerases I and II and Moloney leukemia virus RNA-dependent DNA polymerase by pyran copolymer was dependent on the concentration of the divalent cation cofactor in the reaction mixture. Inhibition was completely blocked by an excess of divalent cations. It was concluded that pyran inhibited these enzymes by complexing with the essential divalent cation cofactor.


Immunofluorescent localization of alpha fetoprotein in yolk sac carcinomas of the rat. alpha-Fetoprotein (AFP) was demonstrated by the immunofluorescent antibody staining technique in 1 primary and 3 transplantable yolk sac carcinomas of rats. AFP was observed only in structures with a characteristic endodermal appearance. This protein was not detected in embryonal carcinoma cells.20


Combined cystometric, sphincter electromyographic and uroflowmetric studies before and after transurethral resection of the prostate. Detrusor and urethral function was studied in 10 patients before and 3 months after transurethral resection of the prostate by  means of flowmetry, and simultaneous gas cystometry and integrated sphincter electromyography. THE PATIENTas cystometry and integrated sphincter electromyography. The patient had no clinical signs of neurological disease. Six had neurogenic exaggeration of the detrusor reflex in the preoperative studies. In 3 patients changes in the detrusor reflex were found postoperatively. Postural changes in detrusor reflex excitability were encountered preoperatively and postoperatively. The preoperative finding of detrusor hyperreflexia in the majority of the patients is ascribed to lesions in the detrusor reflex organization at 2 anatomical sites: 1) a subclinical lesion of the cerebral circuits of the detrusor reflex control owing to arteriosclerosis and 2) an increase of sensory detrusor-reflex triggering stimuli from the morphologically changed prostatic urethra. The study calls for diagnostic techniques for delineation of minimal cerebrospinal impairment and objective assessment of the sensory innervation of the urethra.


The Gibbons indwelling silicone ureteral stent catheter. A method is described to facilitate the placement of the Gibbons indwelling silicone ureteral stent catheter using 2 ureteral catheters. The small catheter is placed within the silicone stent and the larger one is placed over the smaller catheter, fixing the ureteral stent within the obstructed ureter.


[Study of the interdependence between prostatic hypertrophy and disorders in hormone levels. Preliminary report. 1. Study of blood testosterone]. Using the radioimmunological method and applying the double antibody separation technique, testosterone was determined in 14 patients and LH in a few. A group of men in good health aged between 20 and 60 years (22 persons) was used as a control and a group of men aged from 60 to 90 years (24 persons) in whom the size of the prostate gland corresponded to their age. In 4 patients, testosterone levels were determined before and after prostatectomy. It was found that testosterone levels in the serum of men aged over 60, in good health, were markedly lower. Patients with prostatic hypertrophy showed testosterone concentrations several times higher than the men in the control groups. Testosterone levels in patients with considerable hypertrophy of the prostate were higher than in patients with moderate hypertrophy. Adenomectomy had a remarkable influence on the decrease of testosterone in the serum.


Improved chemotherapy in disseminated testicular cancer. Two combination chemotherapy regimens for disseminated testicular cancer are described. Our present regimen of platinum vinblastine and bleomycin has been highly successful, producing 16 complete (80 per cent) and 4 partial (20 per cent) remissions. Furthermore, 2 patients have been rendered free of disease by the surgical removal of residual disease, making the effective complete remission rate in these 20 patients 90 per cent. Of these patients 16 are alive and 14 are free of disease for more than 8 to more than 20 months. Despite the significant toxicity during the first 12 weeks of this therapeutic regimen it usually was manageable and maintenance therapy produced minimal toxicity. We believe that this regimen is a major advance in the management of patients with disseminated testicular cancer.


[Abnormality of the pyelo-ureteric junction revealed by a prostatic adenoma (author's transl)]. An acquired hydronephrosis proximal to obstruction caused by a proastatic adenoma regressed after surgical removal of the latter. On this basis, the authors review the lesions predisposing to abnormalities of the junction and the circumstances which may provoke a hydronephrosis.


Prostatism and prostatectomy: the value of urine flow rate measurement in the preoperative assessment for operation. Urine flow rates were measured before and after elective prostatic operations in 53 patients. The over-all success rate was 72 per cent, as assessed by the changes in symptoms and urine flow rates. Patients with less than a satisfactory operative result had more frequency and urgency and higher preoperative urine flow rates than patients with good results from the operation. Since bladder conditions unassociated with bladder outflow obstruction may be responsible for the poor results it is recommended that urine flow rates be measured before an elective prostatic operation so that patients unlikely to benefit from an operation may be identified.


Radical prostatectomy: palliation for stage C carcinoma of the prostate. An objective comparison is made of patients with stage C carcinoma of the prostate treated with radical prostatectomy versus more conservative measures. Morbidity from local manifestations of the tumor left in situ was markedly increased, whereas those patients afforded an extirpative operation had a much improved quality of life.


Self-instructional emergency medicine program for medical students. A self-instructional program in emergency medicine has been developed for freshmen medical students at the University of Virginia School of Medicine. The cognitive objective of the course is to give the student the minimum level of knowledge to diagnose emergency medical conditions. Performance of the appropriate practical treatment is the course's psychomotor objective. Evaluation of the student's grasp of the program's cognitive and psychomotor objectives is accomplished by the written and practical examination for certification of emergency medical technicians and practical tests in basic life-support. Self-instructional guides, algorithms, videotapes and reading materials are the educational resources for the course. As a result of successful completion of the training program, the student is certified as an emergency medical technician in the Commonwealth of Virginia.


Pathogenesis of polycation-induced alterations ("fusion") of glomerular epithelium. Perfusion of rat kidneys with polycations (protamine sulfate, poly-L-lysine), resulted in glomerular epithelial alterations very similar to those observed in proteinuric states, particularly rat aminonucleoside nephrosis. Such changes did not occur after exposure to neutral or anionic macromolecules (poly-DL-alanine, myoglobin, heparin, poly-L-glutamic acid and ovalbumin). Morphigenetic factors in the polycation-induced lesion included retraction and flattening of foot processes, narrowing of filtration slits, formation of occluding junctions between foot processes and cell swelling. The associated suppression of histochemically demonstrable glomerular polyanion suggested that neutralization of cell surface anionic sites was an important factor in the causation of the lesion, which was reversible by reperfusion with heparin. Observations by freeze-fracture confirmed the similarity of the polycation-induced lesion to the epithelial changes in rat aminonucleoside nephrosis. Following exposure to polycations there was also "staining" of anionic sites on epithelial and endothelial cell membranes and glomerular basement membrane. Reperfusion of protamine-treated kidneys with heparin resulted in restoration of previously suppressed colloidal iron staining and the formation of spherical, electron-dense (heparin-protamine) complexes within the glomerular filter.


Sequelae of concussion caused by minor head injuries. Of 145 patients with concussion from minor head injuries admitted to the Royal Victoria Hospital, Belfast, over one year, 49.0 per cent had no symptoms, 38.9 per cent had between 1 and 6 symptoms, and 2.1 per cent had more than 6 symptoms about six weeks after the accident. There was significant correlation between a high symptom-rate at six weeks and positive neurological signs and symptoms at twenty-four hours. Post-concussion symptoms were more frequent in women, in those injured by falls, and in those who blamed their employers or large impersonal organisations for their accidents. The results suggest that both organic and neurotic factors are involved in the pathogenesis of symptoms at six weeks.


Hyperinsulinism of hepatic cirrhosis: Diminished degradation or hypersecretion? The breakdown of proinsulin in the pancreatic beta cell yields insulin and C-peptide which are secreted in equimolar amounts. Unlike insulin, C-peptide is not degraded significantly by the liver, so that its measurement should give a better assessment of insulin secretion than estimation of peripheral insulin levels alone; particularly in the presence of hepatic dysfunction. Plasma C-peptide and insulin response to an oral glucose load have therefore been assessed in 14 cirrhotic and 7 normal subjects. Cirrhotic patients were divided into hyperinsulinaemic and normoinsulinaemic groups based on fasting plasma-insulin concentrations. Fasting blood-blucose and plasma-C-peptide concentrations were the same in normal and cirrhotic subjects, suggesting that basal pancreatic insulin secretion was the same in all subjects. Thus the C-peptide/insulin ratio was significantly decreased in hyperinsulinaemic subjects (2-13 +/- 0-31, compared with 4-63 +/- 0-48 in controls). After oral glucose, the two groups of cirrhotic patients showed the same glucose intolerance. C-peptide concentrations were also the same but insulin concentrations were markedly increased in the hyperinsulinaemic group. It is suggested that pancreatic insulin secretion is not increased in cirrhosis and that the peripheral hyperinsulinism is due solely to decreased hepatic insulin degradation secondary to either spontaneous portal-systemic shunting or to parenchymal damage.


Autosomal recessive inheritance of susceptibility to tinea imbricata. Familial distribution of chronic tinea imbricata in an untreated Melanesian population was consistent with a genetic predisposition to this disease. The pattern suggested that susceptibility to chronic Trichophyton concentricum infection is recessively inherited and controlled by genes at a single autosomal locus. In married couples there was no concordance of the disease above that expected by chance and the observed segregation of tinea imbricata accorded well with that predicted by the genetic hypothesis.


Subcutaneous fat in newborn infants of diabetic mothers: An indication of quality of diabetic control. Biceps, triceps, subscapular, and suprailiac skinfolds were measured in 40 newborn infants of diabetic mothers. Maternal fasting blood-glucose and mean blood-glucose in the third trimester correlated significantly with neonatal skinfold thickness. Skinfold measurement, when compared with a reference range for gestational age, may be a convenient way of assessing one effect of maternal diabetes on the fetus. Gluteal adipose-cell diameter was measured in 31 infants of diabetic mothers. The fattest babies had the largest adipose cells and there was a significant positive correlation between maternal fasting blood-glucose and neonatal adipose cell diameter. These findings are consistent with the hypothesis that in diabetic pregnancy fetal hyperglycaemia and hyperinsulinism stimulate increased triglyceride synthesis in adipose cells and enlargement of adipose cells and lead to an increase in fetal subcutaneous fat.


Human arterial and venous tissues generate prostacyclin (prostaglandin x), a potent inhibitor of platelet aggregation. Fresh rings of arteries and veins obtained from surgical specimens generated an unstable substance, prostacyclin (prostaglandinx, [P.G.X]) WHICH IS A POTENT INHIBITOR OF PLATELET AGGREGATION. The spontaneous generation of prostacyclin as well as its generation from exogenous arachidonic acid was inhibit by incubation of the tissues with a prostaglandin-synthetase inhibitor such as indomethacin, whilst the generation induced by prostaglandin endoperoxides was not. 15-Hydroperoxyarachidonic acid (a lipid hydroperoxide) inhibited the generation of prostacyclin in all three situations. It is postulated that prostacyclin is important for prevention of deposition of platelets on the vessel wall and that the inhibition or prevention of the generation of prostacyclin is important in the genesis of diseases, especially those in which increased lipid peroxidation occurs, such as atherosclerosis.


Radially oriented fibrin crystallisation. A new test for endotoxin sensitivity in man. Radially oriented acicular crystalline aggregates could be induced by incubating heparinised blood with bacterial endotoxins. These aggregates did not appear in the blood of 37 healthy volunteers but were observed in the blood of 130 patients, predominantly those with vasculitis, psoriasis, and bacterial infections. Study of these asteroid structures, which resemble 'sunbursts', led to the view that they are oriented crystals of fibrin radiating from a central platelet mass undergoing lysis.


Social-services support for multiple sclerosis patients in West of Scotland. A survey has been carries out of 104 patients with multiple sclerosis (m.s.) in West-Central-Scotland. All patients were living at home, and success of services to assist them in the community was examined. The partients were aged 16-65 years and had permanent disability, many being severely handicapped. Regular hospital follow-up was more common among the least disabled. 24 patients had never seen a social worker. Many patients had experienced problems with employment but 37 per cent of these had never registered with a disablement resettlement officer. After advice 10 additional patients applied successfully for an attendance allowance. Legislation requires local authorities to compile a register of the disabled and give information on services available to them. Only 19 patients were registered, however, and no one had received any information from a local authority. Many of these M.S. patients had failed to establish or maintain contact with available services. These results indicate a need for reorganisation of the support for the chronically disabled, possibly by setting up regular clinics for assessment and management, as recommended by the Tunbridge report (1972).


Outcome of pregnancy among women in anaesthetic practice. A survey has been made of the outcome of the pregnancies of 5700 women doctors first registered in England and Wales in 1950 or later. Conceptions that occurred when the mother was in an anaesthetic appointment resulted in smaller babies, higher stillbirth-rates, and more congenital malformations of the cardiovascular system than the pregnancies of other women doctors. There was no significant difference in the spontaneous-abortion rate between the two groups. A pronounced effect of age on this rate was evident among all groups examined.


Oral cimetidine in severe duodenal ulceration. A double-blind controlled trial. 40 adult outpatients with active endoscopically proven duodenal ulceration, who would otherwise have merited elective ulcer surgery, entered a double-blind trial of either cimetidine (1g/day) or placebo. After twenty-eight days, 17 of 20 (85%) patients receiving cimetidine showed ulcer healing, compared with 5 of 20 patients receiving placebo (p less than 0.0005). Patients receiving cimetidine had significantly more pain-free days and pain-free nights than those receiving placebo. There was good correlation between ulcer healing and symptomatic relief (p less than 0.0005).


Neurological and phychometric studies in children surviving freshwater immersion accidents. A study of the neurological and intellectual sequelae of childhood near-drowning is reported. Results are from a total  population study, without selection, of all freshwater immersion accidents in which consciousness was lost in the water. Such accidents affected 56 children in the city of Brisbane and environs over the period 1971-75. 54 of these children have been re-examined medically and psychometrically. Over 95 per cent of children who survived such accidents were neurologically normal. The median i.q. of survivors was 110 (range 90-137), which is higher than that of the general population. There is a suggestion that visualmotor (performance)) skills are particularly vulnerable to freshwater immersion hypoxia. In 20 per cent of survivors subscale disparities between verbal and performance skills exceeded 15 i.q. points. No correlation between the post-immersion I.Q. and either estimated immersion-time or water temperature was demonstrated in this study. No long-term emotional or personality disorders were encountered. Uncommon gross clinical sequelae of prolonged immersion in fresh water included spastic quadriplegia and gross mental retardation. All children in this study were apparently dead at the moment of rescue; despite this, the prognosis of near-drowning in childhood is excellent


1,25-dihydroxycholecalciferol in hypoparathyroidism. 8 patients with hypoparathyroidism have been treated successfully with 1,25-dihydroxycholecalciferol (0-25-1-0 mug/day) for 4-24 mo. Normal serum-calcium levels were restored in all patients, but half the patients required supplementary oral calcium. Treatment reduced serum-phosphorus in all patients, but values remained high in 3, and the renal-tubular reabsorption of phosphate returned to normal in only 2 of the 8 patients. A comparison is made with the effects of oral therapy with massive amounts of calcium salts. The data obtained are discussed in relation to the probable mode of therapeutic action of 1,25-dihydroxycholecalciferol.


Dose-effect relation of cholestryamine in children and young adults with familial hypercholesterolaemia. Twenty children and young adults with familial hypercholesterolaemia (F.H.), on a diet low in cholesterol and high in polyunsaturated fats, were treated with cholestyramine in a metabolic unit to determine the decrease in plasma cholesterol and low-density lipoprotein (L.D.L.) cholesterol in relation to drug dosage, pretreatment concentrations of cholesterol and L.D.L. cholesterol, and body-weight. When the dose of cholestyramine was increased in thirteen patients by 1 g/day up to 16 g/day, given twice daily, cholesterol and L.D.L. cholesterol fell within the normal range in eleven subjects (average dose, 7 g/day), and the response was directly proportional (P less than less than 0-001) to the pretreatment concentrations of cholesterol (r = 0-89) and L.D.L. cholesterol (r = 0-93) but did not correlate with body-weight. Plasma total cholesterol and L.D.L. cholesterol continued to fall and concentrations reached a plateau after which additional cholestyramine had no further effect (average dose, 11 g/day). The L.D.L. cholesterol regression line successfully predicted the dose required to reduce L.D.L. cholesterol concentrations in seven other patients. There was a significant decrease in mean serum-folate in female patients. It was concluded that the minimum effective dose of cholestyramine in young patients with F.H. can be predicted from the pretreatment plasma total and L.D.L. cholesterol and may be given twice daily.


Plasma-exchange and immunosuppression in the treatment of fulminating immune-complex crescentic nephritis. Nine patients with fulminating immune-complex crescentic nephritis were treated by a regimen of intensive plasma-exchange, steroids, and cytotoxic drugs. In five patients with severe renal failure there was early and rapid improvement in renal function; in one patient an early but extensive focal necrotising glomerulitis was arrested; in two patients improvement was delayed for 3 and 7 weeks and could not confidently be attributed to therapy; one patient, anuric at presentation, did not recover renal function. Follow-up renal biopsy specimens, obtained in three patients, showed no evidence of active disease. With the Clq-deviation test, circulating immune complexes were detected in five patients before treatment and had disappeared when renal function had improved and stabilised: these patients showed the best response to therapy. In three patients temporary withdrawal of plasma-exchange was followed by the reappearance of immune complexes in the circulation and was accompanied in two patients by deterioration in renal function; reintroduction of plasma-exchange was followed by elimination of immune complexes and further improvement in renal function.


Genetic factors in the development of chronic active hepatitis. In 14 of 16 patients with chronic active hepatitis (C.A.H.) who did not have HLA antigens B8 and/or B12 an external triggering factor (drug or virus) could be demonstrated at onset of symptoms. In contrast external factors were involved in only 11 of 25 cases of C.A.H. in patients with HLA-B8 and/or B12. In the latter group antinuclear antibodies were less common in cases possible triggered by external agents compared with cases in which no such factor was demonstrated. The results suggest that there are at least two pathogenetically different types of C.A.H.---one genetically determined type in which no external factor is involved and in which autoimmune phenomena are common, and another type triggered by environmental agents and not involving predisposing genetic factors.


Behaviour in ferrets of swine influenza virus isolated from man. After intranasal instillation into ferrets, the "swine" influenza virus A/New Jersey/8/76(Hsw1 N1) had a 50% minimal infectious dose similar to that of previously tested A/PR/8-A/England (H3 N2) recombinants virulent and attenuated for man. A/New Jersey produced only a mild upper respiratory tract infection. However, higher titres of virus were recovered from the lungs over a longer period than experienced previously with Asian and Hong Kong virus strains. There was a diphasic pyrexia the second and higher peaks of which correlated with peak titres of virus in lung macerates. These results suggest that A/New Jersey has a pneumotropic potential in ferrets and, if the animal model is valid, possible in man.


Limitations of transcendental meditation in the treatment of essential hypertension. 20 hypertensive patients participating in a professionally supervised programme of transcendental meditation showed no significant change in blood-pressure after a 6-month study. Although there were small reductions in systolic blood-pressure and in pulse-rate early in the trial, these changes had disappeared by 6 months. At no time did the mean diastolic pressure fall significantly. Plasma-renin activity did not change during the study. It is concluded that while the general feeling of wellbeing experienced by most patients may provide a useful adjunct to conventional treatments, it is unlikely that transcendental meditation contributes directly towards the lowering of blood-pressure.


Myotonia precipitated by propranolol therapy. A 39-year-old man with ischaemic heart-disease developed clinical myotonia while taking propranolol. The myotonia disappeared when administration of the drug ceased. The patient appears to have dystrophia myotonica which had not been evident before propranolol therapy.


Is peptic ulceration a hormonal disease? The pathogenesis of peptic ulceration cannot be explained by an abnormal capacity to secrete acid, for ulcers develop in patients who secrete acid normally. Duodenal and gastric ulcers have a common cause. The location of an ulcer in each individual is primarily determined by his capacity to secrete acid at that time. There is a difference between the mechanisms which heal an ulcer and cure a patient of his disease. Procedures that reduce an individual's capacity to secrete acid, heal an ulcer by moving the focus of the ulcerogenic forces to a more proximal site. It is necessary to remove an antral factor if in addition the patient is to be cured of his disease. It is postulated that this antral factor is the gastrin (G17) which is released in abnormal amounts into gastric juice in patients with ulcers and with gastrinomas. The abnormal amount of G17 in gastric juice may be responsible for releasing abnormal amounts of G34 into the circulation from the duodenum and from gastrinomas. The abnormal release of gastrin develops as a result of an impaired response to duodenal acidification manifest in part by an impaired release of secretin. It is postulated that the abnormal stimulation of antral gastrin release may on occasions give rise to antral G-cell hyperplasia, and that the abnormal secretion of gastrin into gastric juice may on occasions give rise to gastrinomas. These abnormalities may cause ulcers by producing an uncontrolled secretion of acid and an abnormal exposure to bile.


A miniature syringe pump for continuous administration of drugs and hormones: The Mill Hill infuser. A miniature syringe pump has been developed for intravenous or subcutaneous infusion of the many drugs and hormones which produce better therapeutic responses when continuously delivered at a controlled rate than by repeated separate injections. The infuser uses disposable plastic syringes and is designed for precise tamper-proof delivery of 2 ml/24 h or 2 ml/8 h with quartz-crystal control, unaffected bt the dose-rate can be precisely determined by prescribing solutions of appropriate strength.


Abuse of solvents "for kicks". A review of 50 cases. The deliberate inhalation of solvents "for kicks" is becoming more common among children and adolescents in the west of Scotland. 50 consecutive police cases of solvent abuse in Lanarkshire have been reviewed. Investigation of the 42 patients (40 male and 2 female) showed that "sniffing" was a group activity involving many adolescents aged 12-19 years, all of whom had a previous history of solvent abuse. Although some incidents were reported within school hours, 52% of referrals occurred after 6 P.M., with Mondays, Wednesdays, and Fridays being the commonest days for referrals. Of 4 cases sent to hospital, 2 required admission. A list has been compiled of the agents most frequently used for inhalation purposes and a simple screening procedure based on gas-liquid chromatography developed to detect their solvent components. This has been successfully applied to the detection of solvents in blood-samples from the patients referred.


Prolonged exposure to a stressful stimulus (noise) as a cause of raised blood-pressure in man. Systolic and diastolic blood-pressure was significantly higher in 44 male industrial workers with a noise-induced auditory impairment (greater than or equal to 65 dB at 3000, 4000, or 6000 Hz) than in 74 males of the same age with normal hearing. Moreover, significantly more individuals with hypertension (resting recumbent blood-pressure greater than or equal to 160/100 mm Hg) were found in the group with noise-induced loss of hearing. It is suggested that replaced and prolonged exposure to a stressful stimulus (industrial noise severe and prolonged enough to cause a permanent loss of hearing at the relevant frequencies) may be a contributing factor to the rise in blood-pressure through a mechanism involving structural adaption of blood-vessels in response to repeated peaks of raised blood-pressure.


B-lymphocyte alloantigens associated with multiple sclerosis. 6 B-lymphocyte alloantigens have been provisionally identified with lymphocytotoxic antisera reacting, after absorption, specifically with B but not T cells. 3 of these antigens appear to form part of an allelic series. The frequency of HLA and B-lymphocyte antigens was then studied in 59 patients with multiple sclerosis (M.S.). 1 of the B-cell antigens, BT 101, was found in 49 out of 59 patients (83%), compared with 10 out of 30 normal individuals (33%), giving a relative risk of 9-8 times to the association. 2 other B-cell alloantigens and HLA-B7 showed lesser but significant positive associations with M.S. Apart from providing possible clues to the pathogenesis of M.S., the association between BT 101 and M.S. may allow screening for susceptible individuals who are thought to be at special risk.


Effect of intermittent compression of the arms on deep venous thrombosis in the legs. Despite the presence of venostasis in the legs, intermittent compression of the arms during and after surgery reduced the incidence of deep venous thrombosis (D.V.T.) in the legs to half that in control patients and maintained blood fibrinolytic activity at preoperative values. It is suggested that the release of fibrinolytic activators is essential to the prophylactic action of pneumatic leggings.


Treatment of hypersplenism by embolus placement in the splenic artery. In a patient with liver cirrhosis and severe hypersplenism resistent to corticosteroids splenectomy was attempted but proved impossible. Embolisation of the splenic artery with tiny fragments of absorbable gelatin sponge induced gradual restitution of haematological function to normal after 6 wk. Abdominal pain, paralytic ileus of short duration, transient pyrexia, and pleural effusion ensued but were well tolerated. The patient has remained well, 2 1/2 mo later. It is suggested that this simple method may prove rewarding and could safely be used, if necessary, more than once in patients for whom other treatments are unsuitable or have failed.


Reconstitution of B and T lymphocyte function in severe combined immunodeficiency disease after transplantation with thymic epithelium. A patient with severe combined immunodeficiency has been given a transplant of thymic epithelium obtained from short-term culture of normal human thymus. 4 wk after transplantation immunoglobulin was detected and the patient now has normal levels of the three main classes. Functional antibodies of four specificities have been detected. An increase in reactivity to allogeneic cells and phytohaemagglutinin has been observed, with an increase in E rosettes; a positive delayed skin test to candida antigen is now present. The patient has shown reversal of wasting and has been free of infection for 7 months since receiving the transplant. It appears that under certain circumstances thymic epithelium reconstitutes both T and B cell functions.


Islet-cell antibodies in diabetes mellitus. Islet-cell antibodies (I.C.A.) were found in 38% (319/829) of insulin-dependent diabetic patients, in 5% (6/112) of insulin-independent diabetics, and in 1.7% (3/177) of non-diabetic subjects. In the insulin-dependent group I.C.A. were found in 85% of patients immediately after the onset of symptoms and they became less common as the duration of disease increased I.C.A. were equally common in both sexes and the decline in their prevalence was independent of age. The antibodies were directed against cytoplasmic components of islet cells but not against insulin itself. The appearance of I.C.A. probably follows cell damage occurring before the onset of symptoms. By contrast, thyroid and gastric autoantibodies were more common in older patients and females. There was no correlation between the presence of these antibodies and I.C.A. in patients with either diabetes of recent onset or longstanding disease.


Differential response of chlamydial and ureaplasma-associated urethritis to sulphafurazole (sulfisoxazole) and aminocyclitols. 91 men with non-gonococcal urethritis (N.G.U.) were randomly treated with either sulphafurazole (sulfisoxazole), 500 mg orally q.i.d. for 10 days, or an aminocyclitol (streptomycin or spectinomycin), 2 g intramuscularity for 1 to 3 doses at 12 h intervals. Initial urethral cultures were positive for Chlamydia trachomatis (C) in 36 (40%). Ureaplasma urealyticum (U) was isolated from the urethra or urine from20 (95%) of 21 White men in a first episode of N.G.U. who had negative chlamydia cultures. Sulphafurazole, active against C. trachomatis but not U. urealyticum in vitro, produced a clinical response in 7 of 7 men with C+U- N.G.U. and 5 of 19 with C-U+ N.G.U. (P less than 0-01). Aminocyclitols, active against U. urealyticum but relatively inactive against C. trachomatis in vitro produced a clinical response in 0 of 6 men with C+U-N.G.U., 9 of 11 men with C-U+N.G.U. from whom ureaplasma was eradicated (P less than 0-01), and 0 of 8 with C-U+ N.G.U. from whom ureaplasma was not eradicated. C+U+ N.G.U. responded poorly to both antimicrobials alone. These results support the aetiological importance of both C. trachomatis and U. urealyticum in N.G.U.


Subcutaneous infusion and intramuscular injection of desferrioxamine in patients with transfusional iron overload. The effects of intramuscular injection and subcutaneous infusion of desferrioxamine (D.F.) on urinary iron excretion were compared in eleven patients with thalassaemia major and one with congenital sideroblastic anaemia who were being maintained on regular blood-transfusions. Total (48-hour) urinary iron excretion ranged from 3-3 to 40-3 mg (mean 16-3 mg) in nine patients who received 750 mg D.F. intramuscularly before transfusion and from 3-9 to 32-3 mg (mean 11-9 mg) in ten patients who received D.F. by the same route after transfusion. In all 9 patients studied before transfusion, continuous subcutaneous infusion of 750 mg D.F. over 24 hours increased iron excretion by 61-5 to 135-8% (mean 101+/-25-4 S.D.%) compared with intramuscular injection of a similar dose. In the 10 patients studied after transfusion, the iron excretion produced by continuous subcutaneous infusion was from 18-9 to 213% (mean 128+/-74-3%) more than that produced by a single intramuscular injection of D.F. When the subcutaneous dose over 24 hours was increased to 1500 mg in six patients, 48-hour iron excretion ranged from 29-2 to 81-2 mg (mean 52-4 mg) and was increased by 80-2--794% (mean 429%) compared with the excretion when 750 mg was given by intramuscular injection. It is concluded that continuous subcutaneous infusion of D.F. produces more iron excretion in patients with iron overload than intramuscular injection. Providing a suitable portable pump can be carried by the patients, continuous subcutaneous infusion of desferrioxamine may prove a valuable means of preventing or treating iron overload in anaemic patients maintained on regular transfusions.


Bacteraemia in a London teaching hospital 1966-75. Analysis of blood-cultures done at St. Bartholomew's Hospital over the decade 1966-75 has revealed a five-fold increase in the incidence of bacteraemia. This increase was particularly prominent in patients with non-malignant disease. The predominance of bacteraemia caused by gram-negative organisms declined in recent years, whereas bacteraemia due to gram-positive organisms, anaerobes, and Candida spp. increased. A recent increase in the number of blood-cultures performed was associated with the introduction of commercially prepared bottles. This led to a higher rate of investigation of patients who ultimately proved to have negative results, and also to a higher incidence of contamination which exceeded the positivity-rate.


Inappropriate antihypertensive therapy in the elderly. Six symptomless patients aged 64-84 (mean 72) years received antihypertensive therapy from their family doctors. Pretreatment systolic pressures ranged from 160 to 220 mm Hg and disastolic pressures from 80 to 120 mm Hg. Within one week of starting therapy all six patients were admitted as emergencies with epidoses of unconsciousness. Admission systolic pressures ranged from 80 to 150 mm Hg and diastolic pressures from 50 to 90 mm Hg. Before admission each patient had experiences symptoms of postural hypotension and had become housebound. After antihypertensive therapy was stopped, one patient had a residual left homonymous hemianopia but the others recovered completely. A raised systolic and distolic pressure is common in the elderly; potent antihypertensive treatment may seriously impair the quality of life and is often unecessary.


Amputation of the toes for vascular disease: fate of the affected leg. 53 patients coming to amputation of one or more toes for the late results of degenerative vascular disease were studied prospectively. By a median time of thirteen months, 26 of the 53 had undergone a major amputation of the affected side. Diabetes was associated with the same prognosis as atherosclerosis obliterans uncomplicated by diabetes. A palpable pedal pulse or a functioning arterial reconstruction carried a virtual guarantee of success for the toe amputation. The presence of a popliteal pulse, however, was not associated with any better prognosis than the presence of a femoral pulse alone. Smoking seemed to exert little influence. With the passage of time, the major-amputation rate rose steadily, and by 3 1/2 years almost three-quarters of the patients had come to major amputation.


Small-bowel abnormalities in multiple sclerosis. Jejunal biopsies were performed in 12 randomly chosen patients with multiple sclerosis. The jejunal mucosa was examined histologically, ultrastructurally, and by tissue immune techniques. Histology showed a normal mucosa in 7 patients, increased inflammatory-cell infiltration in 3, a partial villous atrophy in 1, and a subtotal villous atrophy in the remaining patient. Fine structural abnormalities were seen in 6 of 8 patients studied. These included microvillous changes, increase in theliolymphocytes and epithelial lysosomes, thickening of the connective tissue with or without collagen fibres, and numerous macrophages containing large amounts of membrane-bound electron-dense material. The latter was seen in 5 of the 8 mucosae examined.


Evidence that endogenous calcitonin protects against renal bone disease. Plasma concentrations of calcium, phosphate, alkaline phosphatase (A.P.), immunoreactive calcitonin (iC.T.), and immunoreactive parathyroid hormone (iP.T.H.) were measured in fifty-two patients with chronic renal failure on maintenance haemodialysis. On the basis of a bimodal distribution of values for plasma-A.P. the patients were dividied into 2 groups. In those patients with normal A.P. concentratons as well as in twenty-eight normal subjects there was a positive correlation between iP.T.H. and iC.T. which was independent of plasma calcium or phosphate. Patients with increased plasma-A.P. had higher concentrations of iP.T.H., lower concentrations of iC.T., and showed a negative relation between the concentrations of the two hormones. It is suggested that a possible factor in the pathogenesis of renal bone disease is a failure to secrete C.T. in adequate amounts.


A simple method of estimating progression of chronic renal failure. In 31 of 34 patients with chronic renal insufficiency caused by various diseaes, reciprocal serum-creatinine concentration declined linearly as creatinine concentration rose from a mean of 2-6 mg/dl to 14-8 mg/dl over an average of 71 months. These results indicate that in most cases reciprocal serum-creatinine declines linearly with time as chronic renal failure progresses. Analysis of this relation in individual patients gives an estimate of the progression of the disease, may help to determine the effects of therapy, and could be used to predict when dialysis will become necessary.


HLA-linked genetic control of host response to Mycobacterium leprae. Non-random parental HLA-haplotype segregation is demonstrated in siblings with leprosy. A new method is described for the statistical analysis of non-random segregation among sibships of different sizes. Sibs with the same type of leprosy show a significant excess of identical HLA haplotypes. This is also true for families in which only tuberculoid leprosy is found, which is by far the commonest type in the population studied. However, sibs affected with different types of leprosy share a haplotype less often than expected. This indicates that both susceptibility to and type of leprosy are controlled by at least two HLA-linked genes. Our findings suggest that the equivocal results of previous population studies are due to differences of linkage disequilibrium between HLA-linked genes controlling the host response to Mycobacterium leprae and alleles of HLA A and B loci in various populations.


Prognostic significance of lymphocyte surface markers in adult non-Hodgkin's malignant lymphoma. Neoplastic tissues from 75 adults with non-Hodgkin's malignant lymphoma were examined for B and T lymphocyte surface markers. All nodular lymphomas were B-cell type. Of 40 diffuse lymphomas, 23 were B, 5 were T, and 12 were "null" type. Patients with nodular lymphoma survived significantly longer than those with diffuse lymphoma (P = 0-00003). For patients with diffuse lymphoma, however, surface markers provide prognostic information not obtainable by histological classification as "poorly differentiated lymphocytic" or "histiocytic". Patients whose malignant cells had B markers survived significantly longer than those whose malignant cells had no markers (P=O-008). Survival of patients with diffuse lymphoma was best predicted by a classification utilising both surface markers and histological appearances. Differences in survival among patients with B-cell and "null" or T-cell lymphomas may relfect differences in sensitivity to specific drugs.


Rapid decrease in plasma-triiodothyronine during surgery and epidural analgesia independent of afferent neurogenic stimuli and of cortisol. Changes in circulating triiodothyronine (T3), thyroxine (T4), binding of thyroid hormones to plasma proteins (resin-T3 test), cortisol, and glucose were evaluated in sixteen patients undergoing abdominal hysterectomy. In eight of the patients afferent neurogenic impulses from the surgical area were blocked during and after operation by epidural analgesia. These patients were pain-free, and the normal stress-induced increase in cortisol and glucose was abolished. During epidural analgesia and general anesthesia plasma-T3 fell rapidly and values in the hypothyroid range were found 6 hours after skin incision. Similarly, an increase in the resin-T3 test reflected decreased binding of T3 to plasma proteins. Plasma-T4 decreased slightly during surgery and epidural analgesia (as it does when other anaesthetics are given), but increased during general anaesthesia. These results indicated that the alterations in thyroid hormones and their binding to plasma proteins after surgery are not caused by a stress-induced increase in plasma-cortisol or by neurogenic afferent stimuli from the surgical area, factors which are both known to affect concentrations of other hormones.


Cell partition. A simple test for lymphocyte sensitisation. An established method of investigating total surface charge on particulate materials has been applied to the detection of changes in surface charge on macrophages which are induced by supernatants from the interaction of sensitised lymphocytes and appropriate antigen. The method was studied as an alternative to the established but problematical macrophage-electrophoretic-mobility (M.E.M.) test for detecting such changes. This preliminary investigation suggests that the results from subjects with and without malignant disease differ significantly.


Mortality from pneumococcal meningitis.

Mortality from pneumococcal meningitis. The records of 207 patients with pneumococcal meningitis admitted to Ahmadu Bello University Hospital, Zaira, northern Nigeria, between February, 1971 and June, 1976 have been reviewed. Mortality was 51%. Death was more likely in patients with a short history, impaired consciousness on admission, and an associated pneumonia. Cerebrospinal fluid levels of protein, lactate, fibrin degradation products, and polysaccharide antigen were higher in patients who died than in survivors.


Outbreak of hospital infection with a strain of Staphylococcus aureus resistant to gentamicin and methicillin. A previously unreported strain of Staphylococcus aureus, resistant to both gentamicin and methicillin, as well as other antibiotics, caused an outbreak of hospital infection involving 17 patinets and 8 staff. In this outbreak none of the patients had received previous aminoglycoside therapy. The epidemic strain was eliminated by the use of strict measures for the control of infection.


Maximum acid output and risk of peptic ulcer. The relationship between the capacity to secrete acid and the risk of peptic ulcer has been examined prospectively in 114 healthy symptom-free students and retrospectively in 2361 patients with and without ulcers. The risk of ulcer was found to increase as te maximum acid output (M.A.O.) increased, and the risk of recurrent ulceration, after vagotomy and drainage for duodenal ulceration, was found to increase as the postvagotomy M.A.O. increased. The risk of recurrent ulcer, at any postvagotomy M.A.O., was always greater than the risk of ulceration in a healthy individual with an equivalent M.A.O.. The addition of an antrectomy to a vagotomy restored the risk of recurrent ulcer towards that of a healthy individual developing his first ulcer. The therapeutic benefit of adding an antrectomy to a vagotomy could not be attributed solely to its enhancement of the percentage reduction in M.A.O. from 65% to 95%. The major therapeutic effect of an antrectomy seems to be achieved independently of its action on M.A.O.


Maximum acid output and position of peptic ulcers. In a retrospective analysis of 2218 tests of gastric secretion 27% of patients with duodenal ulcers had an abnormal capacity to secrete acid. The abnormality was evident only in patients who had had symptoms for longer than three years, and was greatest in patients who had had symptoms for six to nine years. There was no significant difference between the capacity to secretic acid in patients who had symptoms of duodenal ulcer for less than three years and the capacity to secretic acid in normal controls. The tendency for the capacity to secrete acid to increase with duration of symptoms was also evident in patients with gastric ulcers. The positions in which ulcers were found were closely related to the maximum acid output (M.A.O.), and to the age of patients. The site of recurrent ulcers, after vagotomy and drainage, was also related to the M.A.O. after vagotomy. These influences of ageing and vagotomy on the site of ulcers can be attributed to their antecedent effect on the M.A.O. It is suggested that the capacity to secret acid alone is not responsible for the genesis of peptic ulcers but that it influences the position in which an ulcer may develop under the influence of an unknown ulcerogenic factor.


Remission of myasthenia gravis following plasma-exchange. A course of daily plasma-exchange was undertaken in 3 patients with severe myasthenia gravis (M.G.) who had failed to respond to anticholinesterases, thymectomy, and steroids. In the 2 cases with acquired M.G., exchange was associated with an unequivocal improvement in muscle weakness and fatiguability, indicating that a humoral factor in plasma is directly concerned in causing the disorder of neuromuscular transmission. The improvement began within a few days of initiating exchange and continued for several days after exchange ceased. Preliminary evidence suggests that the beneficial effect can be maintained with subsequent immunosuppressive therapy. The patient with congenital M.G. failed to improve, suggesting that its pathogenesis may differ from that of the acquired disease.


Low lead levels and mental retardation.

Low lead levels and mental retardation. Borderline and mildly retarded children attending the hospital developmental evaluation clinic were divided into two groups on the basis of the presence or absence of a pred with a paediatric control group using blood-lead concentration as the independent variable. Children with a history of diagnosed lead posisoning were excluded from the study. The group of mentally retarded children "aetiology unknown" had statistically significantly raised blood-lead concentrations but the mentally retarded sample with "probable aetiology" showed no significant difference in lead concentrations from those of the normal controls. It is concluded that the association between lead and mental retardation extends over a much wider range than hitherto suspected and that the nature of this association is independent of a history of "encephalopathic" lead poisoning. It is suggested that physicians should consider raised lead levels in their examination of all children suspected of mental retardation and that the numerical definition of lead toxicity should be re-evaluated.


Penicillinase-producing Gonococci in Liverpool. Gonococci, which had acquired a TEM-type of penicillinase widely distributed among gram-negative bacilli, appeared in February, 1976, and soon accounted for 9% of isolates at a clinic in Liverpool. In 45 patients infected by such gonococci, the frequency of complications did not suggest reduced communicability or invasiveness, and usual forms of treatment with penicillins always failed. Spectinomycin succeeded in 21 (95%) of 22 patients treated, blt tetracyclines succeeded in only 13 (68%) of 19. Appropriate laboratory tests for recognising penicillinase-producing gonococci must be used since such gonococci have already been transferred to other parts of the U.K. Penicillinase-stable cephalosporins were active in vitro and could prove to be the future treatment of choice.


Red-blood-cell composition in thyroid disease. Eight red-blood-cell (R.B.C.) consitituents were measured in ten patients with hyperthyroidism and in ten healthy subjects. Only R.B.C. sodium ([Na]) and zinc ([Zn]) differed much between the groups. Therefore, only these variables were measured in a larger group of untreated hyperthyroid patients. The increase in. R.B.C. [Na] and decrease in R.B.C. [Zn] were confirmed. The R.B.C. [Na] and [Zn] were related to each other and to the plasma-thyroid-hormone concentration. However, more patients had low R.B.C. [Zn] (91%) than had raised R.B.C. [Na] (50%). Further studies suggest that the R.B.C. [Zn] lags behind the clinical response when these patients are treated. These results suggest that the measurement of R.B.C. [Zn] may have a role in the diagnosis of hyperthyroidism.