Determination of virus-specific antigens in extracts from herpes simplex virus-infected cells by a Cr release inhibition test. A 51cr release inhibition test (CRIT) was developed for determining herpes simplex virus type 1 antigens in infected cell extracts. Among the factors studied were the optimal concentrations of complement, antibody, and infected target cells. Under standardized conditions, the relationship between antigen content and inhibition of chromium release was linear in a semilogarithmic plot, indicating that the antigen content can be determined from testing two dilutions of a given preparation. Other tests showed that infected cells cryopreserved at -70 degrees C for periods exceeding 3 months can be used in CRIT. Five different batches of infected cell extracts were tested in both CRIT and blocking neutralization tests with comparable results being obtained, suggesting that both tests measure the same activity.


Immunogenic properties of the glucosyltransferase from Streptococcus sanguis OMZ 9: kinetic study of inhibition by antibodies. An anti-glucosyltransferase serum was prepared against a pure enzyme preparation from Streptococcus sanguis OMZ 9, which synthesized both soluble and insoluble dextran. Sera, crude gamma globulins, and antibody fractions obtained after gel filtration on a Bio-Gel P200 column were used to study enzyme-antibody interactions. A strong inhibition of glucosyltransferase activity was obtained only with the purified antibody fraction. Kinetics studies showed that the anti-glucosyltransferase antibodies acted as noncompetitive inhibitors with respect to the substrate (sucrose). The addition of primer dextran in the reaction mixture during preincubation produced a diminution of the inhibition, and the antibodies acted as mixed type inhibitors with respect to dextran. The simultaneous addition of dextran and antibodies can protect the enxyme against antibody inhibition.


Specificity of the blastogenic response of human mononuclear cells to herpesvirus antigens. Peripheral blood mononuclear (PBM) cells from donors with a history of prior infection with herpes simplex virus, varicella-zoster virus, and/or cytomegalovirus were cultured for 2 to 8 days with glycine-extracted antigens prepared from these viruses and from infectious bovine rhinotracheitis virus. The proliferative response of PBM cells from all donors was specific during the first 6 days in culture. During this period the cellular immune responses of the seronegative donors were clearly different from those of the seropositive donors. The responses of PBM cells in culture with any of the human herpesvirus antigens studied was not influenced by prior infection of the donor with one or more other human herpesviruses. In contrast, although no donors had antibody to infectious bovine rhinotracheitis virus, the PBM cells from some of them had a proliferative response to this bovine herpesvirus, which increased with time. This nonspecific response appears to be a host-associated function which may be related to recognition of a common herpesvirus antigen.


Purification and partial characterization of hepatitis e antigen (HBeAg). Purification of hepatitis e antigen (HBeAg) from 200 ml of chimpanzee plasma was accomplished by a combination of ion-exchange chromatography on diethylaminoethyl-cellulose followed by gel filtration. High-resolution sodium dodecyl sulfate-polyacrylamide gel electrophoresis of purified HBeAg demonstrated two major polypeptides with estimated molecular weights of 22,000 and 55,000. HBeAg labeled with 125I showed a high affinity for protein A-conjugated Sepharose CL-4B. The precipitation reaction between HBeAg and anti-HBe was inhibited by preincubating the purified antigen with rabbit anti-human immunoglobulin G (IgG). These data show that HBeAg is associated with  a serum fraction with the biophysical and antigenic properties of an immunoblobulin of the IgG class. Sedimentation coefficient analysis of purified HbeAg resulted in an S20w value of 11.6 and a molecular weight value of 324,000. These findings, supported by gel filitration and polyacrylamide gradient gel electrophoresis, revealed that HBeAg has properties analogous to those of a dimer of IgG.


Electron-microscopic study of the anaphylactic degranulation of human basophils from atopic subjects. Leukocytes from subjects allergic to Dermatophagoides pteronyssinus were incubated for 20 min with a solution of D. pteronyssinus extracts. Histamine release was measured at 0, 3, 10 and 20 min. Simultaneously, samples were treated for electron microscopy in such a way as to correlate histamine release and the morphological aspects of basophil leukocytes. The principal features accompanying histamine release were: a progressive activation of the cytoplasmic membrane which showed long processes, densification of the mitochondria, fusion of granulations, progressive dissolution and exocytosis of the contents of the granulations, short segments of rough endoplasmic reticulum, active Golgi apparatus, and thin membrane-bound granules suggesting resynthesis of mediators.


Characterization of the Hanganutziu-Deicher (serum-sickness) antigen as gangliosides containing n-glycolylneuraminic acid. Gangliosides that possess the same sugar sequence but differing in the type of sialic acid (N-acetyl- or N-glycolylneuraminic acid) were compared for their reactivity with Hanganutziu-Deicher ('serum sickness') antibodies by double-diffusion gel precipitation tests. Only N-glycolylneuraminic acid containing gangliosides formed precipitation lines with Hanganutziu-Deicher antibodies, thus suggesting that Hanganutziu-Deicher antigens are gangliosides that contain N-glycolylneuraminic acid.


Comparison between the macrophage electrophoretic mobility (MEM) and the fixed tanned erythrocyte electrophoretic mobility (FTEEM) tests in the detection of cancer. When peripheral lymphocytes from patients with a history of cancer are incubated with encephalitogenic factor (EF), in 90% of cases the resulting products reduce the net surface negativity of guinea-pig macrophages, used as detector cells, as revealed in the macrophage electrophoretic mobility (MEM) test. The MEM test is positive in 36% of people with no history of cancer. Formaldehyde-fixed tanned sheep erythrocytes have been used as detector cells in place of guinea-pig macrophages, in a fixed tanned erythrocyte electrophoretic mobility (FTEEM) test, with lymphocyte products identical to those used in MEM tests. In patients with a history of cancer, positive results were obtained in 28/42 cases with the FTEEM test compared with 32/42 in the MEM test. In people with no history of cancer, negative results were obtained in 16/18 cases with the FTEEM test, compared with 12/18 in the MEM test in the present series, and 51/69 in a more extensive series. These differences are not significant. Cases in which discrepancies are revealed between the two tests are discussed in terms of individual case histories.


Colonies of EBNA-positive cells in soft agar from peripheral leukocytes of infectious mononucleosis patients. Epstein-Barr virus (EBV)-associated nuclear antigen (EBNA)-positive lymphoblastoid cells grew as colonies in soft agar after seeding of leukocytes from the peripheral blood of four patients with infectious mononucleosis serologically determined to be caused by EBV. In individual cases more colonies were obtained from blood specimens during the acute phase of the disease than during the convalescent phase. Incorporation of human umbilical cord serum, which contained neutralizing antibody to EBV, into the agar medium did not reduce the number of colonies developing. Our observations indicate that colony-forming cells were originally present in the blood samples, and that they were not infected and subsequently transformed in vitro. Cells from less than 20% of the EBNA-positive colonies grew to form lymphoblastoid cell lines, which were EBNA-positive and had B lymphocyte surface markers. However, the majority (over 80%) of the EBNA-positive colonies failed to form immortalized cell lines. No colonies were obtained from 91 blood samples from healthy young adults and from five patients with an IM-like disease unrelated to EBV infections. The present results strongly suggest that already transformed cells or cells very easily transformed by EBV are present in the blood of IM patients.


Early increase of serum alpha-fetoprotein in spontaneous hepatocarcinogenesis in mice. The concentration of serum alpha-fetoprotein (AFP) was followed in C3H mice having a high incidence of spontaneous liver-cell cancer. No general elevation of serum AFP level with age was seen in mice without tumor. With a single exception, mice bearing hepatocellular carcinomas had increased serum AFP levels. In some mice this increase followed a biphasic course. Mice killed within 1 month of the time when an elevation of serum AFP was first observed had small tumors or no detectable tumor. Premalignant lesions were present in the livers of 11 out of 16 mice that had elevated AFP but no cancer, while only one out of 14 mice with normal AFP had such alterations. Our results strongly suggest that spontaneous hepatocarcinogenesis proceeds through almost the same premalignant lesions as chemically induced carcinogenesis, and that an increase in AFP production occurs early during this process, often preceding macroscopic lesions. Autologous antibodies to AFP were produced in a group of C3H mice by immunization with rat AFP. These anti-AFP antibodies reduced the amount of serum AFP but had no effect on the incidence of spontaneous hepatomas.


Early detection of tumor-specific antibodies to mammary carcinoma. BALB/cfC3H female mice which develop mammary tumor virus (MTV)-induced mammary tumors possess not only serun antibodies directed against virus-associated antigens which are expressed on mammary tumor cells but also antibodies directed against antigenic specificities which are unique to each tumor. The possible range of unique specificities is considerable; a survey of serum samples from 18 tumor-bearing females revealed only two which reacted with a tumor-unique specificity of another isologous MTV-induced mammary tumor. A retrospective study revealed that in multiparous females, the serum activity against tumor-unique antigenicity could be detected 1 to 4 months before the tumor was grossly visible. On the other hand, in virgin females, the serum activity was detectable only when the tumor was visible. On the basis of this difference in time of origin of reactivity, we speculate that the mechanism of tumor formation in parous females may involve a longer preneoplastic or latent neoplastic period than in virgin females; in the latter the change from normal to overtly neoplastic appears to be more abrupt.


Mammary leukaemia (ML) antigen isolated from L 1210 leukaemia cells. Crude 3 M KCl extracts from L 1210 cells and purified ML antigen were characterized and the presence of ML antigens was verified by various anti-ML sera, as well as by antisera to disrupted MMTV B particles. Selected ML-positive fractions from preparative polyacrylamide electrophoresis and purified ML antigen isolated by affinity chromatography were analyzed by polyacrylamide disc electrophoresis. With both procedures the same sharp band was obtained. The molecular weight of both preparations was found to be 73,500 as estimated. The relationship between ML antigen, MMTV antigenic products and cell-associated viral proteins is discussed.


EBV antigens in lymphocytes of patients with exudative tonsillitis, infectious mononucleosis and Hodgkin's disease. A protocol utilizing isokinetic gradients to isolate human lymphocytes is combined with another that purifies the C3 receptor-bearing B lymphocyte subpopulation, thus enriching the EB virus genome-carrying population. Also, rabbit antisera were prepared to the Epstein-Bar virus nuclear antigen (EBNA) and the early antigen (EA) and utilized in an indirect immunofluorescence test (IIT) to detect these antigens in human lymphocytes isolated from various disease states. Using these methods we demonstrated excellent correlation between standard methods previously employed to detect EB virus-coded antigens and our IIT employing xenogenic antisera. Such tests were done on lymphoblastoid cell lines as well as lymphocytes isolated directly from patients with EB virus lymphoproliferative diseases. Human palatine tonsil-derived lymphocytes from children with exudative tonsillitis and peripheral blood lymphocytes of infectious mononucleosis contained only EBNA in C3 receptor-bearing B lymphocytes. However, patients with lymphoproliferative disorders, including Hodgkin's disease, harbored in their spleens and lymph nodes lymphocytes producing both EBNA and EA.


Presence of common surface antigens(s) on endodermal tumors and embryonal tissues of rats, hamsters and mice. Antiserum against yolk-sac carcinoma of rat was prepared in rabbits. After appropriate absorption in vitro or in vivo this antiserum was examined on different tumors and normal tissues or rat, hamster and mouse. The methods used were indirect immunofluorescence and indirect immunoperoxidase staining and cytotoxicity tests. The immune serum was found to react with the cell membrane of different rat and hamster yolk-sac carcinomas. It reacted also with the cell surface of rat hepatoma cells. By absorption on hyalin and blocking with amniotic fluid it was shown that the antigen was neither a basement membrane component nor alpha-fetoprotein. The antiserum was cytotoxic to yolk-sac carcinoma and hepatoma cells. The immune reaction was limited to the cell membrane, as observed in immunofluorescence and in immunoperoxidase staining. The specificity of the antiserum was proved by cross-absorptions with various tumor lines and by removing its activity with the soluble fraction of yolk-sac carcinoma cells. Non-endodermal rat and hamster tumor lines did not react with the anti-yolk-sac carcinoma immune serum. Most normal adult tissues, including spermatozoa, were negative, but a positive reaction was observed in ovaries and on glandular cells of the uterus. In embryonal tissues this surface antigen(s) was detected in the endoderm of 8-day-old rat embryos 7-day-old mouse embryos and in yolk-sac endoderm of both species. The data indicate that the antigen(s) is associated with endodermal differentiation.


Antigenic differences between AKR lymphoma and thymus cells leading to detection of a tumor antigen associated with immunological enhancement. In an experimental model conditioning for enhancement, an AKR lymphoma was made to grow in BALB/c mice, permitting the simultaneous comparison of tumor-bearing (progressor) and tumor-rejecting (regressor) animals. By immunofluorescence using as target AKR lymphoma and normal thymus cells, both acetone-fixed and unfixed, it was observed that the allogeneic progressor serum contained three antibodies, two of which could be asborbed by thymocytes while the other combined selectively with the acetone-fixed lymphoma target. This tumor-specific antibody could not be detected in regressor serum which, on the other hand, could be completely absorbed by thymocytes. The identification of this acetone-resistant tumor antigen led to the preparation of aceton-treated acellular lymphoma extracts: a precipitate was obtained which upon inoculation in BALB/c mice produced an antiserum that combined selectively with lymphoma targets. In vivo experiments showed that pretreatment with this antigen led to a significant increase in allogeneic tumor incidence, 76% as compared to 37% in the controls. It is concluded that in this allogeneic model, an acetone-resistant tumor-specific antigen and the corresponding antibody are involved in tumor enhancement.


Catalase in salivary gland striated and excretory duct cells. III. Immunocytochemical demonstration with fluorescein and peroxidase-labelled antibodies. There has been disagreement as to the identity of the enzyme responsible for the peroxidate activity in luminal epithelial cells of distal ducts of salivary glands; both peroxidase and catalase could be responsible. Our immunocytochemical investigations using anti-catalase antibodies demonstrate that there are high levels of catalase in these cells in the mouse submandibular gland confirming previous enzyme histochemical studies from this laboratory. Since only relatively small amounts of lactoperoxidase are observed in ductal cells by conventional histochemistry or immunocytochemistry, there can be little doubt that the majority of the peroxidatic activity in striated and excretory duct luminal epithelial cells is due to catalase.


Saccharide distribution in human trophoblast demonstrated using fluorescein-labelled lectins. The reaction of six Flourescein-conjugated lectins with human trophoblast and foetal limb are described. The trophoblast had marked membrane fluorescence after treatment with wheatgerm and Lotus tetragonolobus lectins. Concanavalin A reacted strongly with the trophoblastic cytoplasm but did not react with the nuclei. Most of the lectins had only weak reactions with foetal tissue, apart from the cartilage matrix which fluoresced strongly after treatment with several of the lectins. In addition, concanavalin A reacted with amylase-labile material, probably glycogen, in the skin epidermis and chondrocyte cytoplasm. Various substances were tested in order to determine the nature of the wheatgerm lectin receptor of trophoblast. Several secretory and binding proteins were found not to be responsible, and the identity of the receptor remains unknown.


Ultrastructural visualization of sulphated complex carbohydrates in blood and epithelial cells with the high iron diamine procedure. The high iron diamine (HID) method has been found to impart density at the ultrastructural level selectively to sites known to contain sulphated complex carbohydrates. Thus, immature primary granules in rabbit heterophils, immature precrystalloid granules in rabbit eosinophils, all granules of rabbit basophils, mouse and rat mast cells and the nucleoids of alpha-granules of rabbit platelets were stained by HID. Granules of mast cells in rat cervical lymph node varied in the distribution pattern of the HID-reactive component. Mucous droplets within goblets of mouse colonic epithelial cells varied in HID reactivity. Sites known to contain sialomucin but no sulphates, such as mucous cells and apical plasmalemmae in mouse rectosigmoid colon, failed to stain with HID in contrast to their reactivity of dialysed iron at the ultrastructural level. The surface of mast cells and blood cells lacked affinity for HID, indicating that the dialysed iron binding at the surfaces can be attributed to neuraminic acid. HID proved more effective than dialysed iron in visualizing acid mucosubstance in precursor forms of the crystalloid granules in the eosinophil and in mast cell granules. Inclusion of 0.5% glycerol in the HID solution enhanced staining in mouse colon.


[The usefulness of cryotherapy in ORL surgery (author's transl)]. A report of benefits of cryosurgery and its clinical application based on 125 patients is presented. Cryosurgery was especially appropriate, palliatively and curatively, for lesions of the mouth and pharynx and superficial lesions of facial skin.


Pantomimic representation in psychotic children. Previous research suggests that verbal deficits among psychotic children may be paralleled by deficits in nonverbal pantomime. However, certain questions such as the level of pantomime exhibited by psychotic children, its susceptibility to modification, and its relation to other symbolic functions have not been systematically examined. To investigate these issues, 24 psychotic children were required to represent absent objects (e.g., toothbrush) via pantomime after receiving verbal instructions or instructions accompanied by a model demonstrating the pantomime. Also, measures of receptive and expressive speech, human figure drawings, and pretend play were obtained. The findings indicated very few complete failures in pantomime; higher pantomime performance when a model was provided although even in this condition most responses consisted of low-level substitutions of a body part in place of the absent object; and significant relationships between pantomime and measures of receptive vocabulary, echolalia, drawing, and play. The relationship of the findings to symbolic functioning in normal children and their relevance to understanding symbolic deficits in psychotic children are discussed.


Decreased levels of collagen mRNA in rous sarcoma virus-transformed chick embryo fibroblasts. We have found that double-stranded cDNA synthesized in extended reactions by avian myeloblastosis virus reverse transcriptase is suitable substrate for a variety of restriction endonucleases. Experiments in which rabbit reticulocyte mRNA was reverse-transcribed and restricted to generate beta-globin-specifihe nucleotide sequence of beta-globin mRNA. This method has been applied to study collagen mRNA synthesis in normal and Rous sarcoma virus (RSV)-transformed chick embryo fibroblasts. Characteristic sets of collagen cDNA restriction fragments were produced from RNA fractions rich in collagen message activity. These sets of cDNA fragments, generated by the restriction endonucleases Hae III and Hap II, provided a convenient marker for the presence of collagen mRNA sequences. Equal quantities of high molecular weight mRNA from chick embryo fibroblasts (CEF) and RSV-CEF were reverse-transcribed and the resulting cDNA was restricted. The relative yields of collagen cDNA fragments from such reactions strongly suggest that the decrease in functional collagen RNA following RSV-induced transformation of CEF represents a decrease in the copy number of collagen messenger sequences. The potential of this approach for the study of regulation in other systems is discussed.


Comparison of immunofluorescence and immunoperoxidase staining for identification of rubella virus isolates. To explore possible advantages which immunoperoxidase (IP) staining might have over immunofluorescence (IF) staining for identifying rubella virus isolates, direct comparative studies were done on the same coded clinical materials using the same rubella immune rabbit serum as the primary antiserum in both systems. The rubella immune rabbit serum and conjugated anti-rabbit immune globulins could be used more dilute in the IP system than in the IF system. Both IP and IF staining detected rubella antigen in all specimens which were positive by interference. IP staining also detected low levels of rubella antigen in a few additional specimens which had originally been positive for rubella virus, but which on retesting were negative by interference and IF staining. With second-cell-culture-passage material, IP and IF staining showed comparable specificity, and the few specimens which reacted nonspecifically generally did so in both systems. Cell cultures inoculated directly with urine specimens showed greater nonspecificity by IP than by IF, but this activity could be abolished by pretreatment with sodium azide and peroxide; other methods tried for inactivating endogenous peroxidase activity destroyed rubella antigen as well. The intensity of staining for positive specimens was comparable in the two systems. However, more antigen was demonstrable in both systems when BHK-21 cells were inoculated as a cell suspension and then permitted to grow into monolayers than when the same specimens were inoculated into preformed monolayers. IP staining was considered to be a highly satisfactory alternative to IF staining for identification of rubella virus isolates.


Ontogeny of actin and microsomal antigens in gastric parietal cells. Six fetal and 10 neonatal rat or mouse stomachs and a 14-week human fetal stomach were examined for immunofluorescence reactivity with four sera containing parietal cell antibody (PCA) and four other sera containing smooth muscle antibody (SMA). In rat and mouse stomachs, parietal cells first reacted with PCA in 19-day fetal stomachs and with SMA in two-day neonatal rat stomachs or newly-born mouse stomachs. SMA reactivity with fetal rodent stomachs was restricted to the cytoplasm of smooth muscle, the apices of gastric mucosal cells, and the cytoplasm of fibroblasts surrounding invaginating gastric pits. In the 14-week human fetal stomach, parietal cells stained with PCA but not with SMA. Specificity of the staining reactions was established by the complete inhibition of PCA staining by serum absorption with a gastric microsomal fraction but not with actin. Conversely, the SMA staining was abolished by serum immunoabsorption with actin but not with microsomal fraction. These observations, indicating that the development of the parietal cell microsomal antigen precedes that of actin, may be used to distinguish between the staining of parietal cells by SMA and PCA.


Enzyme immunoassays with special reference to ELISA techniques. In this review outlines are given on various types of enzyme immunoassay. The applications to such enzyme immunoassays, especially ELISA, are dealth with in detail. It is concluded that these techniques have high sensitivity and will be suitable in due course as routine laboratory tests.


The pharmacokinetics of bleomycin in man. A series of studies were carried out to investigate the pharmacokinetics of the antineoplastic antibiotic bleomycin in patients with neoplastic disorders. Drug was administered by long (four to five days) and short (10 minutes) zero-order infusions, and serial plasma and urine samples were collected. Serum and urine bleomycin concentrations were determined by radioimmunoassay. The disposition of bleomycin after the 10-minute infusion was described by a two-compartment open model. However, following multiple-day infusion estimates were obtained that were inconsistent with those from the short infusion. Parameters from the long infusions agreed with those from the short infusion when terminal plasma bleomycin levels less than 10 microunits/ml were excluded. The time to reach steady state following the long infusion (ca. 12 hours) was consistent with the half-life (3 hours) predicted by the short infusion and the long infusion excluding levels less than 10 microunits/ml. Possible explanations include assay interference by an unknown metabolite or strong binding of drug to tissues with release at a rate much less than the apparent rate of elimination of drug from the body.


Computed tomography and transabdominal ultrasound in the evaluation of the prostate. The prostates of eight patients scheduled to undergo prostatectomies were studied prospectively by transabdominal gray scale ultrasound and computed tomography. Computed tomography gave excellent visualization of prostate morphology and pelvic anatomic relationships. Ultrasound provided more histopathologic information in the cases of carcinoma and prostatitis. Additionally, its capacity to distinguish surgical capsule from adenoma, in benign prostatic hypertrophy, made it more useful for volume determinations.


Sources of potential residue problems in dairy herds. Drugs and chemicals provide economic benefit to animal and crop production; to derive this benefit they must be used appropriately. Data from the United States Department of Agriculture, Food Safety and Quality Service residue monitoring program indicates certain drugs and chemicals are not being used appropriately by the dairy industry, and residues are occurring in cow carcasses at slaughter. Residues of antibiotics and certain pesticides which are administered intentionally are being found. Dairy cows are also particularly susceptible to inadvertant exposure to chemicals because of certain management practices and physiologic characteristics of dairy cows. Contamination of Michigan dairy cows with polybrominated biphenyls is an example. Methods of controlling both intentional and unintentional exposure of cows to drugs and chemicals will require considerable involvement of producers.


The direct demonstration of histamine release in allergic reactions in the skin using a skin chamber technique. We have adapted a skin chamber technique to permit sampling of fluid at the skin window sites of pollen antigen-induced allergic reactions. Low background levels of histamines are formed in control chambers, whereas significantly increased (p less than 0.01) amounts are found within 30 min following ragweed application in sensitized subjects.


The effect of aldehyde fixation on selected substrates for energy metabolism and amino acids in mouse brain. The effect of aldehyde fixation on concentrations of low molecular weight constituents was determined by comparing amounts of selected intermediates in brains of mice exposed to aldehyde fixative solutions with those perfused with phosphate buffer solution alone. Aldehyde perfusion resulted in excellent preservation of cerebral cortex ultrastructure in the presence of dramatic declines in adenosine triphosphate, phosphocreatine, glucose and glucose-6-phosphate that occureed before exposure of the tissue to aldehyde fixatives. Decreases in hexose were accompanied by approximately a 4-fold increase in lactate and a 2-fold increase in pyruvate. Glycogen levels decreased by about 60% during the initial operative procedure but remained constant after aldehyde fixation. Glycogen content declined approximately 90% in tissues that were not treated with aldehyde. Concentrations of aspartate and glutamate changed only slightly during the initial period (1-5 min) and remained constant for at least 90 min in cerebral cortices fixed with aldehydes. Alanine levels increased in both fixed and unfixed tissue; however, this increase was much smaller in tissues exposed promptly to aldehydes. Total ninhydrin-positive material in perchloric acid extracts of brain decreased in mice exposed to aldehyde solutions but increased in tissues that were not. These results indicated that several amino acids may be measured reliably in tissues preserved for light and electron microscopy. In addition, determination of glutamate: alanine ratios in tissues perfused with aldehydes may provide an indication of the timing of fixation.


Genetic analysis of Ia determinants expressed on Con A-reactive cells. Pretreatment of mouse lymphoid cells with anti-Ia sera and C abrogated the proliferative responses of these cells to Con A. Studies were carried out with several anti-Ia reagents and intra-H-2 recombinant mouse strains to map the I subregion(s) whose products are expressed on Con A-reactive cells. Treatment with a (B10.A X A)F1 anti-B10 reagent and C abrogated the ability of BALB/c cells to respond to Con A. Absorption studies on this reagent demonstrated that Con A-reactive cells express Ia determinants coded by the I-A subregion. The results with two additional reagents, B10.A(4R) anti-B10.A(2R) tested on B10.BR cells and (B10 X D2.GD)F1 anti-B10.D2 absorbed with B10.A cells and tested on BALB/c cells, demonstrated that Con A-reactive cells also express Ia determinants encoded to the right of I-A. Several antisera and strain combinations were evaluated in which the antisera could contain antibodies specific for products of genes encoded by the I-J subregion, but the results were inconclusive. These data demonstrate that there are at least two different I subregions, one in I-A and one to the right of I-A, that code for antigens expressed on Con A-reactive cells.


Separation and partial characterization of a type-specific antigen from Chlamydia trachomatis. Type-specific antigens of Chlamydia trachomatis have been demonstrated by the mouse toxicity prevention test and a variety of immunofluorescent techniques. In addition, biologic activity has been associated with these antigens in terms of type-specific immunity to trachoma infections. This report is the first to describe the detection of a soluble type-specific antigen of C. trachomatis and its separation from those antigens that cross-react among different immunotypes. Test antigens were prepared by labeling the surface components of purified, yolk sac grown organisms with a radioiodinated intermediate (Bolton-Hunter reagent, 125I). The organisms were solubilized with Triton X-100 and gel filtered through Sepharose 6B. All fractions were then tested in radioimmunoassay for binding with rabbit antisera raised against solubilized immunogens prepared from homologous and heterologous strain organisms propagated in BHK-21 cells. A fraction demonstrating homologous binding only was used in subsequent modified procedures for the preparation of quantities of type-specific antigen sufficient for analysis. The antigen appears to be a heat labile, cell surface protein associated  with apparent immunogenic activity during the course of actual chlamydial eye infection.


In vitro secondary MLR. III: Hybrid histocompatibility determinants. With cells obtained from long-term mixed lymphocyte culture (MLC) we have have demonstrated hybrid histocompatibility determinants that are recognized as mixed lymphocyte reaction (MLR)-stimulating determinants. Hybrid determinants can also be detected (1) in an in vitro assay for effector cells that measure inhibition of immunoglobulin secretion by LPS-blasts, and (2) in DTH-like responses of in vitro-primed cells injected into "B" mice and challenged in vivo with stimulator cells. The possible implications of these findings for the physiologic function of MLR determinants are discussed.


Theory of equilibrium binding of symmetric bivalent haptens to cell surface antibody: application to histamine release from basophils. We present a theory of equilibrium binding of symmetric bivalent haptens to cell surface antibody in the presence or absence of monovalent hapten. Bivalent haptens can link together antibodies to form linear chains or rings on cell surfaces. We show how to calculate the amount of any complex of bound bivalent hapten, monovalene fraction of antibody involved in complexes made up of two or more antibodies, i.e., the fraction of antibody that is cross-linked (Xpoly). We treat the case when the antibody on the cell surface, which is specific for the hapten, is homogeneous. For this case we prove a number of general properties about Xpoly: 1) Xpoly approaches zero at both high and low bivalent hapten concentration. 2) Xpoly becomes a maximum when the bivalent hapten concentration equals Amax, where Amax = 1/H + B/2. H is twice the equilibrium constant for the binding of a single hapten site to a single antibody site and B is the monovalent hapten concentration. 3) a plot of Xpoly vs the log of the bivalent hapten concentration is symmetric about the maximum value of Xpoly. We use these and other properties of Xpoly in this paper to clarify the relationship between cross-link formation and histamine release.


Histamine release due to bivalent penicilloyl haptens: control by the basophil plasma membrane. We describe the characteristics of in vitro histamine release from human basophils passively sensitized with serum from a penicillin-allergic individual. The histamine release is induced by a synthetic bivalent hapten, bis benzylpenicilloyl 1,6 diaminohexane (BPO)2. We present data on the effect of a monovalent hapten, benzylpenicilloyl formyl-L-lysine (BPO)1, on the histamine release. We also examine how histamine release depends on the concentration of serum used for passive sensitization, the source of cells used for passive sensitization, and the time allowed for histamine release. We interpret these experiments in terms of a theory of equilibrium binding of bivalent haptens to cell surface antibody that is presented in the previous paper. The results are consistent with the idea that the amount of histamine release is controlled by the number of cross-linked IgE molecules on the cell surface. In particular, the histamine dose-response curve rises because cross-links rise, has a maximum because the cross-links are a maximum, and falls because the cross-links fall.


Intra- and interspecies reactivity of human and mouse natural killer (NK) cells. Natural killer (NK) cells can kill certain syngeneic, allogeneic and xenogeneic tumor targets in short-term 51Cr release assays. In the present study, intra- and interspecies NK activity was analyzed. Ten mouse and five human tissue culture lines were used. In direct cytolytic assays with mouse spleen cells or human PBL effectors, intraspecies was much stronger than interspecies reactivity, as a rule. A certain interspecies activity was obtained, stronger in mouse anti-human (M alpha H) than in human anti-mouse (H alpha M) combinations. In the H alpha M system, activity was associated with the same type of PBL-derived non-B-non-T cell fraction as in the intraspecies H alpha H system. The non-B-non-T cell nature of the M alpha H killer cell has been demonstrated previously. Nonlabeled tumor cells were allowed to compete with isotope-labeled targets in intra- and interspecies cytolytic NK tests. NK-sensitive tumor lines of the same species were superior to xenogeneic competitors in both M alpha M and H alpha H tests. In the M alpha M assay, the competing ability of the same human tumors varied, depending on the genotype of the mouse effector cells. None of the human lines tested competed effectively with strain CBA effectors but some showed a certain competition with C57BL effectors. In the H alpha H assay, strong competition was seen with two of the 10 xenogeneic mouse tumors tested.


Structural studies of the protein portion of the H-2-linked Ia glycoprotein antigens of the mouse: tryptic peptide comparison of products from the I-A and I-C subregions of B10-HTT. Ia antigens from the I-A8 and I-Ck subregions of the B10.HTT (H-2t3) strain of mice were isolated by indirect immunoprecipitation of arginine-labeled, nonionic detergent-solubilized materials. After biochemical purification the electrophoretically homogeneous 28,000 dalton glycoprotein beta chains from the Ia precipitates were digested with trypsin and the resultant radiolabeled tryptic peptides were compared by analytical ion exchange chromatography. These comparisons reveal that the beta chains of Ia antigens from the A (I-A8) and C (I-Ck) subregions of B10.HTT share only two out of 12 to 14 of their arginine tryptic peptides. Thus these noncross-reactive Ia antigens are structurally quite diverse, and would possess sufficient structural variability to account for their lack of antigenic cross-reactivity.


Characterization of the proteolytic fragments of bovine colostral IgG1. Bovine colostral IgG1 was subjected to both papain and pepsin hydrolysis. Papain digestion appeared to be optimal at pH 7.4 in the presence of 0.01 M cysteine. The molecule was split at the COOH-terminal side of the interchain disulfide bond(s), and in addition to Fab fragments, two Fc fragments, designated Fc(I) and Fc(II), were obtained. Both Fc fragments had an identical NH2-terminal sequence, but differed in m.w. by about 10,000, with Fc(II) being the smaller one. Differences were also observed in their circular dichroism (CD) spectra and in their susceptibility to carboxypeptidase hydrolysis. These results suggested that the distinguishing characteristics of the two Fc fragments resided in the COOH-terminal parts of the molecules. Pepsin hydrolysis yielded the expected F(ab')2 and pFc' fragments. This hydrolysis was found to be dependent upon substrate concentration leading to aggregate formation at IgG1 concentrations below 3%.


An isotope-release assay and terminal-labeling assay for measuring cell-mediated allograft and tumor immunity to small numbers of adherent target cells. A 51Cr-release assay and terminal 51Cr-labeling assay for measuring cell-mediated immunity to adherent target cells is described. Both techniques utilize small 10 microliter-per well microtiter plates, require low numbers of target cells (50-500 per well), and consequently, relatively small numbers of effector cells per well (3x10(3) -1x10(5)). Both assays are objective, quantitative, and simple to perform. The suitability of these techniques for monitoring immunologically specific, cellmediated, cytotoxic response to syngeneic and allogeneic tumor cells and normal skin fibroblasts is demonstrated. Lymph node cells, spleen cells and peritoneal exudate cells serve as effectors.


Technical improvements in the mixed antiglobulin rosetting reaction with consequent demonstration of high numbers of immunoglobulin-bearing lymphocytes in viable preparations of human peripheral blood. A modification of the mixed antiglobulin rosetting reaction (MARR) to improve sensitivity as a test for immunoglobulin (Ig)-bearing human blood lymphocytes is described. A mean 5.7%, lymphocytes Ig-positive by the MARR when rosettes were formed in medium containing 0.2% bovine serum albumin (BSA), increased to 20% when rosettes were formed in 5% BSA or by incubating the lymphocytes or indicator erythocytes with Vibrio cholerae neuraminidase before rosetting. Under these various rosetting conditions the MARR is as sensitive as the direct antiglobulin rosetting reaction (DARR). Further, with the MARR, false positive rosette formation due to unusual antimembrane antibodies can be excluded during the mixed antiglobulin rosetting procedure by use of blocking controls. Substitution of F(ab)2 antiglobulin for IgG anti-gamma, anti-alpha and anti-mu did not reduce the number of lymphocytes demonstrable with the MARR, indicating that the MARR does not react with adsorbed Ig molecules on lymphocytes. Summation of the number of sheep erythrocyte (E) rosetting lymphocytes and mixed antiglobulin rosetting lymphocytes approximated 100%, yet in T-enriched preparations a maximum of 4% of lymphocytes were Ig-positive by the MARR, suggesting that null lymphocytes which have been reported to be E-negative and immunofluorescence-negative are B lymphocytes with surface Ig determinants.


Group-specific component [Gc] levels in chronic liver disease. Serum Group-specific component (a probable vitamin D transport protein) concentrations have been measured in 72 patients with chronic liver disease. Low mean values were found in groups of patients with cirrhosis and metastatic liver disease. In a group of patients with biliary tract disease the mean value was not significantly different from normal except for seven patients with severe bone disease who were found to have the lowest levels. The mechanism for the reduction remains to be clarified, but low Group-specific component values may play a contributory role in the osteodystrophy of chronic obstructive liver disease.


Immunological studies of T-cell receptors. I. Specifically induced resistance to graft-versus-host disease in rats mediated by host T-cell immunity to  alloreactive parental T cells. The present studies extend our previous efforts to understand the immunological basis of specifically induced graft-versus-host (GVH) resistance in F1 hybrid rats. Immunization of F1 rats with alloreactive T-cell populations of parental strain origin induces a host-mediated T-cell response which is specific for anti-major hostocompatibility complex receptors on parental T cells. This protective immunity is rapid in onset and once induced, it provides a highly effective, specific resistance to lethal GVH disease which is radioresistant and can be adoptively transferred to syngeneic recipients.


T-cell populations specifically depleted of alloreactive potential cannot be induced to lyse H-2-different virus-infected target cells. Mouse lymphocyte populations of one parental H-2 type (A) were specificially depleted of alloreactive potential by filtration through irradiated A X B F1 recipients, and thoracic duct cells were then stimulated with virus in an A X B F1 environment. Experiments using T cells that had previously been exposed to influenza virus in the context of A established that cross-priming for recognition of viral components expressed on H-2-different (B) target cells does not occur. Furthermore, immunologically naive T cells stimulated with vaccinia virus, subsequent to negative selection for reactivity to B, could not be shown to interact with virus-infected cells of type B. Either there is no significant T-cell repertoire for recognition of virus associated with an H-2 determinant not encountered during ontogeny, or such T cells are also alloreactive and are removed during filtration.


Simultaneous complementary idiotypic responses: absence of reciprocal regulation. Complementary antibodies, i.e. antibodies having combining site structures which are at least partially directed against each other, were induced in A/He mice by immunization with phosphorylcholine (Pc) coupled to keyhole limpet hemocyanin or with the Pc-binding IgA myeloma protein, HOPC-8 (H8). Both responses were monitored by enumerating plaque-forming cells and assaying serum antibody levels to Pc and H8. Prior immunization with H8 markedly suppressed subsequent immunization with Pc and vice versa; neither plaque-forming cell response was diminished, however, when mice were immunized simultaneously with Pc and H8. Experiments were designed to determine if the absence of reciprocal regulation was due to change in idiotypes. This was determined by measuring inhibition of plaque formation using complementary antibody. Plaque formation by cells was equally inhibited by high dilutions of the appropriate complementary antibody whether cells were from mice immunized with one, the other, or both antigens. Thus, the absence of regulation could not be accounted for by emergence of different idiotypes. Interestingly, sera from mice immunized to have high responses to both antigens were relatively ineffective in inhibiting plaque formation or suppressing immunization to Pc. However, such sera contained complexes of the complementary antibodies; apparently antibody to Pc in such sera quenches or neutralizes the activity of anti-H8 antibody. But the formation of complexes, at least measurable levels of circulating complexes, must be a result rather than the cause for the absence of reciprocal regulation, since regulation was also absent when immunization to Pc was manipulated so that responses were too low to result in detectable levels of circulating antibody to Pc. It is proposed that simultaneous complementary responses may occur in nature to other antigens and antibodies, and that such simultaneous responses may cause pathologic changes.


The detection of a spleen focus-forming virus neoantigen by lymphocyte-mediated cytolysis. The existence of a nonvirion tumor-associated cell surface antigen (TASA) on cells transformed with Friend (FLV) on Rauscher (RLV) leukemia virus has been difficult to demonstrate. Antisera raised against classically defined Friend- Moloney-Rauscher antigenic determinants have been shown to react with virus structural proteins coded for by genetic information contained in the lymphatic leukemia or helper (LLV) virus genome. The recent development of nontrans-formed fibroblast cell lines which contain the replication-defective spleen focus-forming virus (SFFV) genome, free of replicating LLV, has allowed investigation of an SFFV-specific antigen. We have applied the techniques of mixed tumor-lymphocyte culture stimulation followed by lymphocyte-mediated cytolysis assays to search for the cell surface expression of an antigen coded expressly by SFFV genetic information. SFFV nonproducer-immune, in vitro activated spleen cells were capable of effecting the lysis of SFFV-containing BALB/c 3T3 and Fischer rat epithelial, cloned cell lines. Normal BALB/c 3T3 and BALB/c 3T3 cells infected with three types of ecotropic LLV were unaffected. Syngeneic FLV and RLV-induced murine leukemia cells were also killed by SFFV nonproducer-immune lymphocytes. In addition, Kirsten sarcoma virus-transformed, replication-defective and replication-rescued BALB/c 3T3 fibroblasts were not susceptible to SFFV antigen-directed cytolysis. Antibody-dependent complement-mediated cytolysis assays using monospecific goat antisera confirmed that SFFV nonproducers lacked cell surface expression of virion structural proteins. These observations suggest that the antigen detected in LMC experiments was not coded for by genetic information contained in the helper component of FLV, and that it represents a true SFFV-specific cell surface antigen. Based upon the recent molecular evaluation of the SFFV genome as consisting of both xenotropic and ecotropic virus sequences, it appears reasonable that xenotropic genetic information may be responsible for expression of the SFFV- specific antigen. Since the replication-defective SFFV genome is also responsible for the malignant transformation associated with FLV-induced erythroleukemia, one might postulate that gene sequences capable of programming transformation may also code for the TASA detected in these studies.


A study of proteases and protease-inhibitor complexes in biological fluids. We have (a) screened a variety of cell lines and body fluids for plasminogen activators and (b) studied the activity of proteases bound to alpha2- macroglobulin after exposing the complexes to partial degradation and/or denaturing procedures to unmask proteolytic activity. The respective results show (a) that the plasminogen activators in urine and cell culture media are generally of lower molecular weight than those in plasma; and (b) that proteases bound to alpha2-macroglobulin recover the ability to attack macromolecular substrates after exposure to sodium dodecyl sulfate while retaining the electrophoretic mobility of the protease inhibitor complex. This indicates that the protease and inhibitor are probably linked by covalent bonds. In contrast, other complexes formed between proteases and inhibitors of lower molecular weight (such as soybean or Kunitz inhibitors) are fully dissociated by sodium dodecyl sulfate (SDS). The experiments described were based on a new procedure for detecting proteolytic enzyme activity in SDS-polyacrylamide gels. The method relies on solutions of nonionic detergents for extracting SDS, after which the electrophoretic gel is applied to an indicator gel consisting of a fibrin- agar mixture. The method is sensitive, permitting the detection of proteinases in less than 1 mul of fresh plasma, and it is effective for resolving small differences in molecular weight. The procedure can be quantitated and, with minor modifications appropriate to each particular system, it has been applied to a broad spectrum of serine enzymes and proenzymes, including some that function in the pathways of fibrinolysis, coagulation and kinin-generation. Other potential applications appear likely.


Evidence for a pathogenic role of a cell-mediated immune mechanism in experimental glomerulonephritis. Lewis rats were injected intravenously with rabbit anti-rat glomerular basement membrane (GBM) antisera in doses that were sufficient to cause glomerular fixation of rabbit gamma globulin (RGG) detectable by immunofluorescence, but which failed to induce histologically detectable lesions. 24 h later, groups of rats received lymph node cells or serum from syngeneic donors that had been immunized with either RGG or ovalbumin; they were injected with [3H]thymidine three times during the next 2 days, and sacrificed 48 or 96 h after transfer. Only the rats given anti-GBM antiserum plus lymph node cells from donors sensitized to RGG showed histological glomerular lesions, in the form of segmental hypercellularly and necrosis. Autoradiographs revealed the greatest number of labeled cells in glomeruli in the same group. In analogous experiments, it was shown that T-cell-enriched populations could induce hypercellular glomerular reactions. On the basis of electronmicroscopic and autoradiographic observations, it appears that the glomerular hypercellularity resulted from both infiltration of mononuclear cells and proliferation of endothelial cells. The findings indicate that interaction of specifically sensitized lymphocytes with glomerular-bound antigen can induce a cell-mediated (delayed-type) reaction in glomeruli.


Involvement of fusion activity of ultraviolet light-inactivated Sendai virus in formation of target antigens recognized by cytotoxic T cells. Mice inoculated with ultraviolet light-inactivated Sendai virus mount a cell- mediated immune response to the virus. Cytotoxic T cells specific for Sendai virus can be obtained by in vitro secondary stimulation of primed spleen cells with syngeneic stimulator cells coated with UV-inactivated Sendai virus. Neither in vivo nor in vitro stimulation alone is sufficient to generate specific cytotoxic T cells. Sharing of the H-2 haplotype between cytotoxic T cells and target cells is required for the Sendai virus-specific lysis to occur. The fusion (F) glycoprotein of Sendai virus has been implicated in target antigen formation (20). Ethanol treatment of Sendai virus causes complete inactivation of the cell-fusion and hemolytic activities of the envelope, but does not affect the antigenicity of the F glycoprotein; furthermore, hemagglutinin and neuraminidase activities of the envelope HANA glycoprotein are also left intact after ethanol treatment. Target cells can be prepared by coating them with various numbers of UV-inactivated Sendai virus that have been treated with ethanol or, as a control, phosphate-buffered saline (PBS). The amount of virus adsorbed to target cells during the cytotoxicity reaction time using either ethanol-treated or untreated (PBS "treated") virions is essentially identical, but target cells coated with ethanol-treated Sendai virus fail to serve as targets for cytotoxic T cells. These results indicate that fusion activity of the Sendai virus envelope is essential to the formation of the target antigen and that virus adsorption to cell surfaces without fusion of the envelope with cell membranes is not sufficient to allow killing by virus-specific cytotoxic T cells.


Interspecies spleen-myeloma hybrid producing monoclonal antibodies against mouse lymphocyte surface glycoprotein, T200. A cell hybrid has been selected from fusion of a mouse myeloma and rat spleen cells immunized against mouse lymphoma cells that produces monoclonal antibody against the mouse lymphocyte surface glycoprotein, T200. Antibody binding assays employing the monoclonal antibody show that there are about 50,000-100,000 molecules of T200 glycoprotein on mouse thymocytes and that similar antigens are present on spleen and bone marrow but not detected on nonlymphoid tissues. Examination of the labeled molecules precipitated from detergent extracts of spleen cells and thymocytes iodinated by the lactoperoxidase technique by SDS-PAGE confirm that there are structural differences between the antigens found on B and T lymphocytes. The B-cell glycoprotein consists of at least one component of apparent mol wt 220,000 on SDS-PAGE, while the T-cell glycoprotein has an apparent mol wt of about 190,000.


In vitro maturation of immature thymocytes into immunocompetent T cells in the absence of direct thymic influence. Peanut lectin (PNL) binds to a majority of mouse thymocytes (Thc) in suspension. By using cell affinity chromatography on a column of anti-PNL antibody, Thc populations at least 96 percent pure in PNL + or - cells, as judged by immunofluorescence, were obtained. PNL(+) cells are rich in Thy 1 and poor in H(2) antigens, cortisone sensitive, unresponsive to phytohemagglutinin (PHA), and immunologically incompetent, as judged by mixed lymphocyte reaction, popliteal lymph node graft-versus-host assay, and by testing helper activity in a primary in vitro antibody response to sheep erythrocytes; the converse is true of PNL(-) cells. Thus, PNL(+) and (-) cells appear to correspond to cortical and medullary Thc, respectively, as previously suggested. In culture, PNL(+) Thc show poor viability and a weak proliferative response to concanavalin A (Con A), except when supernate (SUP) of 24 h Con A stimulated lymph node lymphocyte cultures, or irradiated lymph node cells, are added, in which cases a strong proliferative response to the mitogen is observed. A variety of control experiments showed that the proliferating cells did not result from preferential stimulation of a few contaminating PNL(-) Thc present in the PNL(+) Thc cultures. The blasts resulting from PNL(+) Thc proliferation display mitogen-induced cytotoxicity, and give rise to a population of medium-sized lymphocytes, mostly PNL(-), poor in Thy 1 and rich in H(2) antigens, PHA responsive, and immunologically competent in the above-mentioned assays. Fresh PNL(+) Thc responded in mixed lymphocyte reaction in the presence of SUP (lectin depleted) and since incubation in SUP alone did not confer reactivity on PNL(+) Thc, it appears therefore that (a) immature Thc possess alloantigen and mitogen-specific surface receptors but lack the capacity to respond by proliferation to receptor triggering without the help of extracellular factor(s) released by mature lymphoid cells stimulated by mitogens (b) cell division is associated with the acquisition of immunological responsiveness, characteristic of mature T lymphocytes. The implications of these findings for the ontogenesis of thymus-derived lymphocytes, and for the possible traffic of Thc within and from the thymus, are discussed.


Association of the H-Y male antigen with beta2-microglobulin on human lymphoid and differentiated mouse teratocarcinoma cell lines. The expression of the H-Y antigen has been tested on several human lymphoid lines and mouse teratocarcinoma cell lines during differentiation. The human male lymphoid cell line Raji is a very useful target for studies of the H-Y antigen by lymphocytotoxicity test with rat anti-H-Y sera. With a few exceptions, all cells carrying the Y chromosome were H-Y positive. One of the exceptions is the human Daudi cell line which, besides lacking H-Y antigen, also lacks beta2-microglobulin. We have studied a possible association between the H-Y antigen, beta2-microglobulin, and HLA antigen with redistribution experiments. The results strongly suggest that H-Y antigen is not associated with HLA antigens but with beta2-microglobulin.


In a fully H-2 incompatible chimera, T cells of donor origin can respond to minor histocompatibility antigens in association with either donor or host H-2 type. Fully H-2 incompatible radiation chimeras were prepared using BALB congenic mice. Such chimeric mice were immunized in vivo against histocompatibility antigens of the C57BL/10Sn (B10) background in association with either of the parental H-2 haplotypes, and their spleen cells subsequently boosted in vitro with the same minor antigens. Strong H-2-restricted cytotoxic activity against minor antigens was detected, and the specificity of the restriction could be to the H-2 haplotype of the donor or the host depending on the cells used for priming or boosting. Cross priming could also be demonstrated in these mice. The results show that fully allogenic radiation chimeras can produce H-2-restricted T-cell responses to minor histocompatibility (H) antigens, and are discussed in relation to contrasting results recently obtained against viral antigens.


Relationship of human adenoviruses 12, 18, and 31 as determined by hemagglutination inhibition. Adenoviruses 12 and 31, but not Ad18, agglutinate rat blood cells at high titer, providing suitable blood cells be available and a prolonged contact period of virus with the erythrocytes is allowed. Purified virus particles show direct, and virus-free supernatants show direct and indirect, hemagglutination, ie, enhancement of HA by heterologous antiserum. Hemagglutination inhibition with rabbit antisera shows cross-reactions between Ad12 and Ad31 with titers 4--32 times lower than with homologous antigens; Ad18 antisera react with antigens from both of the other serotypes. No cross-reactions were seen with antisera from other adenoviruses. This suggests an antigenic relationship of the three viruses of subgroup IV in their fiber antigen gamma, in addition to the known relation in the hexon (epsilon), which is apparent in cross-neutralization.


[Diagnosis of acute intermittent porphyria. Results of neurological, biochemical and genetic studies]. Neurological and biochemical studies have been performed on four AIP families with 21 members. Five patients suffered from manifested AIP (Uroporphyrinogen Synthase defect and characteristic urine findings); among their relatives five persons with latent AIP were detected and eight carriers of the genetic-enzymic defect (Uroporphyrinogen Synthase defect). Internal and neurological symptoms could be interpreted as a panneuropathy. Acute and chronical polyneuropathies could be observed as well as myelopathies and cerebral co-reactions. A frequent symptom dominating the crisis and the latent state of AIP were etiologically abscure 'myalgias.' The character of the course of AIP is various and dubious: beyond the 'classical' courses with its intermittent porphyric crises we observed one case which was characterized by a permanent crisis and a second case marked by a chronical, slow progredient course without any porphyric attacks.


Inflammatory disease of the brain diagnosed by computed tomography. Inflammatory disease of the brain, particularly of pyogenic etiology, may be most accurately assessed by the use of computed tomography and contrast enhancement. Examples of cerebritis, evolving and mature intracerebral abcesses, and inflammatory extracerebral collections are presented and discussed, particularly with reference to differential diagnosis and the problem of infectious versus neoplastic etiology. Ancillary studies, particulary selective arteriography, are reviewed where appropriate. The residual changes of inflammatory disease of the brain after medical and/or surgical therapy are illustrated, and the value of serial examinations is emphasized. The rural indigent patient population (American Indian) served by the University of New Mexico Medical Center is a source of case material providing several examples of severe and untreated intracerebral infectious processes heretofore not described in the literature.


In vivo interaction of anticonvulsant drugs. The mathematical correlation of plasma levels of anticonvulsant drugs in epileptic patients. The phenytoin plasma levels were measured in 45 epileptic patients whose only treatment was phenytoin. The plasma of 20 other patients receiving both phenytoin and phenobarbital was also tested for concentration of these two drugs and 18 patients treated with phenytoin, phenobarbital and primidone were investigated in the same way. The results were used to calculate the plasma levels of phenytoin in relation to dosage and to measure the effect of the simultaneous use of phenobarbital on the phenytoin plasma levels and of primidone together with phenobarbital on phenytoin concentration. The results led to the following conclusions: The population of epileptic patients can be divided into 2 groups. In the first group the patients reach equilibrium at the relatively high phenytoin plasma level for a given dose of phenytoin, and in the second group the phenytoin plasma level tends to be significantly lower for parallel dosages. Both groups, in their behavior, obey mathematically an exponential graph specific for each group. Phenobarbital tends to lower the plasma phenytoin level when the two drugs are used simultaneously. It is also possible, by the graphs produced, to calculate the expected phenytoin plasma levels when using the drugs together. Primidone and phenobarbital together decrease the phenytoin level much more than expected from the effect of phenobarbital alone.


Intramedullary syndrome due to an extradural neurinoma near the foramen magnum. A case of right sided extradural neurinoma at the level of the foramen magnum is reported which presented as an intramedullary spinal cord syndrome. Ischemia of the anterior spinal artery or of the vertebral artery was considered to be an important pathogenic factor in the production of the neurological syndrome. The outer part of the spinothalamic tract, where sensory fibers carrying pain and thermal sensibility from the sacral segments are situated, escaped ischemia as that part is supplied by penetrating branches of the pial arterial plexus. The discrepancy between the level of neurological deficit (C5) and site of the tumor (C1-2) was due to distant ischemia. The lack of a history of root pain and the rapid recovery following removal of the tumor also favor a vascular origin for the neurological deficit.


Serum-CSF protein gradients, the blood-GSF barrier and the local immune response. For the majority of proteins there is a steady state equilbrium between the serum and the CSF compartment which depends upon the hydrodynamic radii of the passively transferred molecules. For clinical purposes the serum-CSF concentration ratios of albumin (Aalb) and alpha2-macroglobulin (Qalpha2 M) have proven to be a reliable barrier parameter, which is more sensitive than the total protein level in certain diseases, e.g. disk protrusions, degenerative processes and metabolic disorders. The immunoglobulins G and A cope with the passive transfer mechanism in both normal conditions and all degrees of pure barrier impairments but deviate in cases with local immunoglobulin production. The method described produces a quantitative differentiation between the locally synthesized and the serum-derived immunoglobulin fractions. A humoral immune response within the central nervous system was found in certain stages of acute infectious diseases and with chronic inflammatory processes such as subacute sclerosing panencephalitis, neurolues and multiple sclerosis.


Brainstem lesion with coma for five years following manipulation of the cervical spine. A woman, aged 38, lost consciousness immediately after manipulation of the cervical spine and remained in coma for nearly 5 years in a combined decorticate and decerebrate state. Cerebral angiography revealed impaired circulation in the vertebrobasilar system. The EEG initially showed generalized cerebral dysrhythmia but tended, over the years, to become more normal with desynchronized fast activity of low voltage. Neuropathological examination postmortem revealed a large cystic lesion in the pressure equalization area of the carotid and basilar circulation in the upper pons, mesencephalon, posterior hypothalamus and basal thalamus. No definite stenosis or occlusion was seen in the cerebral vessels on angiography or at the postmortem examination. The mechanism of the lesion is considered to have been temporary interference with the blood flow in the vertebrobasilar system during manipulation of the cervical spine sufficient to cause ischemia and subsequent infarction of the brainstem. The pathogenesis of vascular lesions of the brain stem following manipulation of the cervical spine is discussed briefly.


Effect of low lysine diet on rat protein metabolism. Prolonged lysine deficiency resulted in inhibition of growth of the rat and in lowered levels of albumin and beta-globulins, but not of total liver proteins. The apparent incorporation of [3H]lysine into total and chromosomal liver proteins, however, was increased in lysine deficient rats relative to pair-fed controls. The concentration of free lysine in neither serum nor liver was altered in lysine deficient rats. The specific radioactivity of liver free lysine 2 hours after administration of [3H]lysine was noticeably elevated in the deficient rats. However, when the pool of liver free lysine is considered, synthesis of all proteins studied was inhibited. The largest inhibitory effect was seen in serum albumin and total liver protein synthesis. Inhibition of snythesis of nuclear chromatin protein was less pronounced. The data indicate an adaptive mechanism operates to preserve liver lysine in rats fed lysine deficient diets for a prolonged period. In spite of retention of lysine, the synthesis of liver and serum proteins was inhibited.


Reflex neurovascular dystrophy in childhood. Reflex neurovascular dystrophy has rarely been recognized in children. During the past eight years we have observed 24 instances of RND in 23 children. Lower extremity involvement was manifested in 20 of them and upper extremity in four. The major complaint was pain; swelling and vasomotor instability were prominent, and exquisite tenderness was characteristic. Chronic trophic changes were not observed. Antecedent illness or trauma could be related to the RND in less than half of the children, but personality factors appeared contributory to the development of RND in most children. Physical therapy was the principal form of treatment; therapy with a corticosteroid or by sympathetic blockade was not employed. Reduction in the evidences of disease, including improvement in function, were present in all children at the termination of therapy; improvement was maintained in all but one child after a mean period of 2.4 years. The excellent response to conservative therapy suggests that RND may be a more benign condition in children than in adults.


Systemic lupus erythematosus with negative LE cells and antinuclear factor. In the course of a prospective study of 165 patients with SLE a subgroup of eight patients with active SLE yet with persistently negative tests for ANF and LE cells was identified. These patients were characterized by a photosensitive skin rash with a negative lupus band test, a high incidence of arthritis, mild form of renal disease, and a positive family history for connective tissue disease. Antibodies to DNA and cytoplasmic antigens were detected in a few.


Fucosyltransferase activity in metastasizing and nonmetastasizing rat mammary carcinomas. Fucosyltransferase levels in 6 established strains of spontaneously metastasizing rat mammary tumors (STMT-058, MT-449, DMBA-4, SMT-077, TMT-081, and SMT-2A) were compared with 4 nonmetastasizing strains (MT-W9B, MT-W9A, MT-100, and MT-66) as controls. Two acceptors were prepared from fetuin for the assay, one by acid hydrolysis of N-acetylneuraminic acid and the other by the stepwise removal of N-acetylneuraminic acid and penultimate galactose by Smith degradation. The enzyme that transfers fucose to the first acceptor was designated fucosyltransferase A, whereas the one that uses the second acceptor was designated fucosyltransferase B. Both types of fucosyltransferases were found in this rat mammary tumor system. Whereas the levels of fucosyltransferase A in the 2 tumor groups were comparable, those of fucosyltransferase B were sixfold to sevenfold higher in the metastasizing tumors. This difference in the level of fucosyltransferase B was not caused either by differential hydrolysis of GDP-fucose by pyrophosphatase in the 2 groups or by hydrolysis of the product by fucosidases. Presence of any other inhibitor(s) or activator(s) of fucosyltransferase was excluded by mixing experiments. Optimal conditions for the assay of this enzyme were determined in a representative strain from each group. Under all circumstances, the activity of fucosyltransferase B was higher in the metastasizing tumors. The enzyme was inhibited by nucleoside diphosphates and triphosphates, and guanosine nucleotides were the most efficient inhibitors. Subcellular distributions of the two fucosyltransferases were similar, 35-50% of the enzyme activity being present in the crude microsomes. When plasma membrane factions were prepared from the microsomes, the major part (50-70%) of the enzyme was associated with the light and heavy plasma membrane fractions. Increased activity of fucosyltransferase B in the group of metastasizing tumors may have reflected faster synthesis and shedding of fucose-containing glycoprotein antigens. Similar molecules possibly were also synthesized in the nonmetastasizing cells but at a much slower rate, because the antigen is not easily lost from the cell surface. Any alteration of the specificity of this focosyltransferase in the metastasizing tumors, in addition, may have caused antigen modulation.


Polyproteins related to the major core protein of mouse mammary tumor virus. The mouse mammary tumor virus (MuMTV) contains several low-molecular-weight proteins which, together with the genomic RNA, constitute the core structure of the virion. The most abundant protein in the core is the 27,000-dalton protein (p27), and, by analogy to the type C viruses, this protein probably forms the core shell. In mouse mammary tumor cell lines (GR and Mm5MT) producing MuMTV the major p57 antigenic specificity resides in a large protein, which migrates in polyacrylamide gels as a doublet of 77,000 and 75,000 daltons (p 77/75). A series of lower-molecular-weight proteins, p61, p48, p38, and p34, is also present in small amounts and is probably derived by proteolytic cleavage of the p 77/75. These proteins have been identified by immunoprecipitation with monospecific antiserum, and their sequence relatedness to p27 has been determined by an analysis of the peptides after trypsin digestion. After a 15-min pulse with [35S]-methionine, all of the p27-related proteins in these cell lines were labelled and, during a subsequent chase, progressively disappeared. The p27 was labeled poorly during the pulse, but the amount of label in this protein increased during the chase. A quantitation of these experiments suggested that the majority of the p27-related proteins were quite rapidly turned over in these cell lines. Hence, if p27 is derived by a progressive proteolytic cleavage mechanism, then the process is inefficient in the GR cells and only moderately efficient in the Mm5MT cells. When MuMTV was isolated from the culture medium of these cells harvested at 5-min intervals, the major p27-related protein was p34. The p27 accounted for only 29% of the anti-p27 serum immunoprecipitable proteins compared to 95% in virus isolated from an 18-h harvest. Incubation of the rapid-harvest virus at 37 degrees C for 2 h resulted in some conversion of p34 to p27. These results suggest that some of the p27 in MuMTV is formed in the virions by proteolytic cleavage of p34.


Identification of a common antigen of herpes simplex virus bovine herpes mammillitis virus, and B virus. In immunoelectrophoretic analyses one common antigen was demonstrated in antigen preparations from herpes simplex virus types 1- and 2- (HSV-1 and HSV-2), bovine herpes mammillitis (BHM) virus-, and B virus-infected cells solubilized by Triton X-100. The antigen was also demonstrated in solubilized purified HSV-1 and BHM virus. The common antigen was identified as antigen 11 of HSV-1 or HSV-2. Differences were found in the polypeptide composition of the related antigens when isolated from the four different herpesviruses, but a glycopolypeptide with a molecular weight of 125,000 was present in each of the four different antigen preparations, indicating that this polypeptide carried the common antigenic determinants.


Relationship of retrovirus polyprotein cleavages to virion maturation studied with temperature-sensitive murine leukemia virus mutants. Murine leukemia virus mutants ts3 (Moloney) and ts24 (Rauscher) both formed late-budding structures on the cell membrane at restrictive temperature. They both accumulated core polyproteins Pr65gag and Pr180gag-pol in cell membranes, but the envelope precursor was rapidly turned over. After shift to permissive temperature in the presence of cycloheximide, the accumulated precursors were sequentially cleaved via discrete intermediates both during the final stages of the budding process and in newly released virions to yield the finished virion core proteins and reverse transcriptase. The precursor form of reverse transcriptase was not enzymatically active and became activated partially or entirely inside released virions.


Identification of an FMR cell surface antigen associated with murine leukemia virus-infected cells. FMR antigens are found on the surface of cells infected with Friend, Moloney, and Rauscher murine leukemia viruses (MuLV). These antigens are serologically distinct from the G cell surface antigens that are found on cells infected with endogenous MuLV (AKR and Gross virus). Cell surface antigens of both virus groups are immunogenic in mice, and immunization with appropriate virus-infected cells leads to  the production of cytotoxic antisera. The cytotoxic activity of FMR antisera can be absorbed by disrupted preparations of Rauscher MuLV, but not by AKR MuLV. FMR antisera precipitate the viral envelope proteins gp70, pl5(E), and p12(E) from detergent-disrupted preparations of [3H]leucine-labeled MuLV. The reaction of these antisera with p15(E) and p12(E) proteins is directed against group-specific antigens and can be absorbed with AKR MuLV; in contrast, the reaction of these antisera with gp70 is directed against type-specific antigens and is absorbed only by viruses of the FMR group. In immune precipitation assays with detergent-disrupted 125I surface-labeled cells, FMR antisera react only with type-specific antigens of the viral envelpe protein. On the basis of these findings we conclude that the FMR cell surface antigen is a determinant on the MuLV env gene product.


Multimodal treatment of advanced testicular tumor with radical reductive surgery and multisequential chemotherapy with cis platinum, bleomycin, vinblastine, vincristine and actinomycin D. Advanced testicular tumors in 34 patients were treated by combination chemotherapy with bleomycin, vinblastine, vincristine, cis platinum and actinomycin D. The therapy was divided into 3 phases: 1) induction, 2) consolidation and 3) maintenance. Induction lasted 4 weeks and consisted of 420 mg. bleomycin, 0.2 mg./kg. vinblastine, 4 mg./kg. cis platinum, and 20 mg. prednisone daily. Consolidation lasted 6 weeks and consisted of 5 mg. actinomycin D, 6 mg. vincristine and 6 mg./kg. cis platinum. Maintenance therapy was achieved with 2.5 mg. actinomycin D every 6 weeks and 1 mg./kg. cis platinum every 3 weeks. A tumor reductive operation was done before induction of chemotherapy in 13 patients and after induction of chemotherapy in 12 patients. Nine patients were treated with chemotherapy alone. Three patients with brain metastases received concomitant radiotherapy to the brain (3,000 rads). A previous operation and chemotherapy had failed in 11 patients and previous radiotherapy had failed in 1 patient. All patients treated had at least 1 objective response (34 of 34 or 100 per cent). Partial clinical remission was achieved in 7 of 34 patients (21 per cent). A complete clinical remission was observed in 27 of 34 patients (79 per cent) and of this group 6 had a relapse. At present, 22 of 34 patients are free of disease from 4 to 24 months, with an average of 13 months (65 per cent). The toxicity consisted of nausea, vomiting, mucositis, alopecia, mild leukopenia and tinnitus. This approach seems to be effective in producing long clinical remissions in the majority of patients with advanced disease.


The natural history of metastatic renal cell carcinoma: a computer analysis. Survival factors of 86 patients with metastatic renal cell carcinoma were studied by computer analysis. Cumulative survival was 53 per cent at 6 months, 43 per cent at 1 year, 26 per cent at 2 years and 13 per cent at 5 years. Survival was influenced favorably by confinement of metastases to the lungs, by the absence of local recurrence or persistence of tumor and by a longer interval free of disease after removal of the primary tumor. Medical therapy improved survival during the first year after diagnosis of metastases but no objective regression of tumor was observed. Excision of metastatic foci significantly improved survival for up to 5 years (p less than 0.05 and p less than 0.02) after which most patients died of recurrence. Palliative or adjunctive nephrectomy in patients with metastases was associated with a 6 per cent mortality rate but it increases survival over other patients with metastases at the time of diagnosis of renal carcinoma who did not undergo nephrectomy. This difference was owing to patient selection and survival of  those who had adjunctive nephrectomy was no greater than that of the study population as a whole. However, based on the factors that were associated with improved survival palliative nephrectomy may be beneficial when a limited number of metastases treatable by excision or radiation therapy are present, when effective systemic therapy exists or when the primary tumor produces severe symptoms.


Palliative urinary diversion for malignant ureteral obstruction. An analysis of 62 palliative urinary diversions for malignant ureteral obstruction is presented. The average postoperative survival was 187 days. Cell type, duration of known disease, tumor grade and stage, renal function and previous therapy did not strongly influence survival. Renal function returned to normal in 64 per cent of the azotemic patients. Morbidity and mortality rates were high, largely because of underlying disease and adjuvant therapy . Nearly two-thirds of the patients left the hospital and this group subsequently spent 84 per cent of their remaining survival time at home. A criterion is presented for patient selection and suggestions are made for the selection of an operative procedure.


[Relationship between ventricular extrasystole and parasystole]. Monomorphic ventricular extrasystoles, no less than 30 within an hour, were revealed in 89 of 300 patients with chronic ischemic heart disease during continuous 24-hour ECG recording on magnetic tape. A fixed type of the linkage interval (oscillations of no more than 0.1 sec) was established in 58 (62%) and the variable tupe (oscillations of more than 0.1 sec) in 31 (38%) patients. Oscillations in the linkage interval of more than 0.1 sec gave grounds to suspect parasystole. A deeper analysis revealed parasystole in 8 cases (2.6% of the total number of patients), the remaining cases were included in the group of extrasystoles with a variable non-parasystolic type of the linkage interval. This group of extrasystoles occupies an intermediate position between para- and extrasystole. The large number of transitional forms between extra- and parasystole suggests their common origin.


[Importance of the physical load test for detecting heart rhythm disturbances in patients who have had a myocardial infarct]. The article discusses the data on the detection of ventricular extrasytole by means of the load test on a bicycle ergometer in 150 patients who had suffered from myocardial infarction. Much more patients with latent disorders of rhythm are detected by the load test than by means of the standard ECG recorded at rest. It was noted that the ventricular extrasystole became more detectable with the increase in the rate of cardiac contractions, in the degree to which the maximum pulse during the load test exceeded the pulse at rest, in the systolic arterial pressure, and with the raise in the level of the load. Patients in whom extrasystole was revealed and the test was discontinued before reaching a high level of cardiac rhythm had more complicated disorders of rhythm and the prognosis in such cases was poorer.


[Corneal changes in patients on dialysis for renal failure (author's transl)]. The corneas of 100 patients on dialysis for renal failure were observed. In 52 patients follow up studies were possible. 55 patients demonstrated limbal changes. On slit lamp biomicroscopy these changes showed to be similar to the white limbus girdle described by Vogt. Small limbal biopsies from five patients on dialysis were studies histopathologically and compared to biopsies from three patients with normal renal function. Only patients on dialysis showed senile elastosis and three of them epithelial and subepithelial hyaline deposits. Calcium deposition was the same in both groups.


[Systemic and biochemical manifestations of hepatoblastomas (author's transl)]. Among 220 children with malignant solid tumors diagnosed at the University Children Hospital and Department of Pediatric Surgery in Graz six (1,8%) hepatoblastomas were found. All patients were boys. Systemic manifestations may precede the discovery of the abdominal mass and may be important for diagnosis and follow-up.


Electrophoresis and immuno electrophoresis in leprosy. 50 cases of leprosy belonging to various subgroups i.e. 10 tuberculoid, 25 lepromatous leprosy, 10 lepra reaction, 5 dimorphic leprosy, and 25 cases or normal individuals were subjected to agar gel electrophoresis. The slides were scanned by densitometry. It showed profound departure from normal in various fractions of electrophoretic patterns. All of them showed rise of gamma globulin. Albumin was markedly decreased in lepromotous leprosy and lepra reaction. In dimorphic leprosy Alfa-1 was decreased. Qualitative immuno electrophoresis was done by using antihuman serum raised in the laboratory, by immunising rabbits. It revealed changes in IgM and IgG arcs. Results are discussed and tried to explain on immunological derangement.


Increased dopamine-receptor sensitivity in schizophrenia. Dopaminergic mechanisms have been investigated in post-mortem brain specimens from nineteen patients with schizophrenia and nineteen controls. Dopamine turnover was not increased in schizophrenic patients but, as assessed by the spiroperidol-binding technique, there was a significant increase in postsynaptic receptor sensitivity. The change in the dopamine receptor occurred in nucleus accumbens, putamen, and caudate nucleus. Increased dopamine-receptor sensitivity was present in five patients who had been free of neuroleptic medication for at least 1 year before death, and therefore may be related to the disease process.


Oral vancomycin for antibiotic-associated pseudomembranous colitis. Nine patients with antibiotic-associated pseudomembranous colitis were treated with oral vancomycin. All had severe diarrhoea, tissue-culture evidence of a clostridial toxin in stool, and typical lesions on sigmoidoscopic examination, despite discontinuation of all antimicrobials for periods of 10 days to 8 weeks. Oral vancomycin was given in doses of 2 g daily. All patients showed a good clinical response with gradual resolution of diarrhoea over 7 days and a rapid decrease in concentrations of the toxin in stools. Follow-up sigmoidoscopies in seven patients showed major improvement or complete clearing of lesions after 7-10 days of vancomycin treatment. The mean concentration of vancomycin in twenty-five stools obtained during treatment was 3100 microgram/g, levels in serum being very low. These results suggest a role for oral vancomycin treatment of antibiotic-associated pseudomembranous colitis which persists for extended periods despite discontinuation of the incriminated antimicrobial.


Effect of physiotherapy on pulmonary function. A laboratory study. 33 patients with acute exacerbations of chronic bronchitis were studied within 4 days of admission to hospital. Physiotherapy produced an acute rise in lung volume and conductance, without altering specific conductance. There was no consistent change in arterial-blood gases.


Comparison of endoscopic retrograde and intravenous cholangiography in diagnosis of biliary calculi. The results of endoscopic retrograde cholangiography (E.R.C.) and intravenous cholangiography (I.V.C.) were compared in patients undergoing surgery for biliary calculi. E.R.C. gave a significantly higher rate of correct diagnosis of gallbladder calculi (p = 0.026) and common bileduct calculi (p = 0.002) than I.V.C. Biliary stones may thus frequently be present in patients with a normal I.V.C. and E.R.C. may reduce the rate of diagnostic errors in patients with symptoms of biliary disease.


Persistent Epstein-Barr virus infection in a child with hypergammaglobulinaemia and immunoblastic proliferation associated with a selective defect in immune interferon secretion. A 5-year-old girl had a chronic disease characterised by fever, lymphoid hyperplasia, interstitial pneumonitis, thrombocytopenia, and polyclonal hypergammaglobulinaemia. Evidence for severe, persistent Epstein-Barr virus (E.B.V.) infection was found: titres of antibody to E.B.V. viral capsid antigen (IgM and IgG) and early antigen were extremely high, cells containing E.B.V.-associated nuclear antigen (E.B.N.A.) were found in lymph nodes and blood, and spontaneous permanent lymphoblastoid cell lines were established from both sources over a period of a year. After exacerbation of the polyclonal proliferation of immunoblasts the patient died 19 months after the onset of the disease. No defect in humoral or cellular immunity was detected, except for a selective defect in immune interferon secretion by peripheral mononuclear cells. Our results suggest an important role for immune interferon in host defence against E.B.V. infection and in the regulation of immune responses.


Use of oral contraceptives, cigarette smoking, and risk of subarachnoid haemorrhage. In a large cohort of women, the risk of subarachnoid haemorrhage was significantly associated both with cigarette smoking and with use of oral contraceptives (O.C.S.). The risk of cigarette smokers was 5.7 times that of non-smokers; for current users of O.C.S. the risk was 6.5 times that of non-users. The risk in women who both smoked cigarettes and used O.C.S was 22 times that of women who neither smoked nor used O.C.S. Past use of O.C.S was also significantly associated with increased risk of subarachnoid haemorrhage, the relative risk being 5.3. There was some evidence that risk may increase with duration of O.C. use. In this series no significant associations were found between occurrence of subarachnoid haemorrhage and history of hypertension or of migraine headache.


Abnormal levels of prostaglandins and fatty acids in blood of children with cystic fibrosis. 12 children with cystic fibrosis (C.F.) had lower levels of the essential fatty acid, linoleic acid, in plasma and in red blood cells, than did control children, and production of prostaglandin F2alpha (P.G.F2alpha) was higher than in controls. After 10 months of oral linoleic-acid supplementation in 6 of the children with C.F., the linoleic-acid levels in plasma, red cells, and platelets were higher and P.G.F2alpha production was lower than in the 6 children with C.F. who received a placebo lipid. Prostaglandin F2alpha is associated with bronchoconstriction and its increased production in children with C.F. might be causally related to their chronic pulmonary disease.


Increased unsaturated transcobalamin II in active autoimmune disease. Measurements of transcobalamin II (T.C. II) concentrations in 26 patients with lupus erythematosus, 4 with dermatomyositis, 2 with autoimmune haemolytic anaemia, and in 40 immunosuppressed renal-transplant patients showed that T.C. II levels were raised during active phases of autoimmune disease. Changes in T.C. II levels correlated better with the clinical course of autoimmune disease than did changes in C3, the titre of antinuclear antibody, or native D.N.A.-binding capacity.


Does bile promote extra-colonic cancer?

Does bile promote extra-colonic cancer? The incidence of biliary-tract cancer and of several other gastrointestinal tumours is increased when conditions favour conversion of bile acids to more active substances. This may indicate that bacterial degradation of bile salts is related to the pathogenesis of cancer in parts of the alimentary tract other than the large bowel.


Effectiveness of neonatal screening for congenital dislocation of the hip. Despite the examination of neonates for congenital dislocation of the hip, this condition is being diagnosed after the newborn period at a rate not very different from that reported before the introduction of screening. The majority of late-diagnosed cases had been examined after birth, and the sensitivity of the tests used routinely is called into question. Re-examination of all infants at 3-6 months is proposed to reduce the number of missed cases and so minimise late sequelae.


The paranoid syndrome.

The paranoid syndrome. Paranoid characteristics often prompt the diagnosis of schizophrenia. This paper advises a cautious diagnostic approach aimed at increased awareness of the many and potentially serious illnesses that may present with paranoid features.


Comparison of sucrose with glucose in oral therapy of infant diarrhoea. In a randomised double-blind trial, 51 5--10% dehydrated infants were rehydrated with oral electrolyte solutions containing sucrose or glucose. Most infants in both groups were successfully rehydrated, but the sucrose solution produced a slower correction of electrolyte abnormalities and a higher percentage of patients who needed more than 24 h of therapy. Where there is adequate knowledge of the oral therapy method sucrose can substitute for glucose in many cases; where there is a choice glucose is recommended.


Low-dose insulin infusions in diabetic patients with high insulin requirements. Six patients with high insulin requirements (range 120-3000 units daily) have been infused with much smaller doses (range 50-63 units daily) of insulin intravenously. All six maintained adequate glucose homoestasis on this regimen. It is suggested that subcutaneous tissue at the site of injection may alter insulin or impair its absorption. Insulin resistance in some patients may be due to these mechanisms.


Spinal-fluid procoagulant activity: a sensitive indicator of central-nervous-system damage. Testing the procoagulant activity (P.C.A.) of cerebrospinal fluid (C.S.F.) by measuring its effect on plasma-recalcification time is a useful indicator of central-nervous-system damage. C.S.F. from 22 normal children and adolescents aged 6 months to 17 years had a mean +/- S.D. P.C.A. of 14 +/- 6%. P.C.A. in 13 children with C.N.S. infection was significantly increased to 59 +/- 13%. In 8/13 of these children activity remained high (42 +/- 11%) after therapy. In patients with acute lymphatic leukemia (A.L.L.) 12 aged greater than 2 1/2 years and diagnosis had a normal activity (17 +/- 8%) and 4 patients aged less than 2 1/2 years at diagnosis had an activity of 21 +/- 8%; during C.N.S. prophylaxis with intrathecal methotrexate and/or cranial irradiation, mean activity in these 16 patients rose significantly to 41 +/- 11%. More than 2 years after treatment P.C.A. decreased towards normal in the older children but remained high in the younger group. 5 children with neurological sequelae (including 3 with A.L.L. and the post-irradiation syndrome) had the highest activities. Ether extraction showed that the active material had lipid properties. P.C.A. did not correlate with protein, lactic dehydrogenase, or cell count in C.S.F.


Increased arterial-tissue cholesterol after intestinal bypass in severe obesity. Cholesterol content was measured in biopsy specimens of mesenteric artery taken at the time of jejuno-ileal bypass from 4 very obese patients aged 19, 22, 27, and 42. When their body-weight had become stable (mean 22 months) after operation, biopsy specimens of the temporal artery were obtained from 3 patients and of a mesenteric artery from 1. Cholesterol content had increased significantly. The increase in arterial-tissue cholesterol was concomitant with decreases in serum high-density (H.D.L.) and low-density (L.D.L.) lipoprotien cholesterol.


Noma neonatorum: Its aetiopathogenesis.

Noma neonatorum: Its aetiopathogenesis. Noma neonatorum, a gangrenous process affecting the nose, lips, mouth, anal region, and occasionally the scrotum and eyelids, affects neonates, especially low-birth-weight and premature ill babies, and is usually fatal 1-3 days after onset. In 35 cases of noma neonatorum Pseudomonas aeruginosa was isolated from blood-culture (86.3%), gangrenous areas (96.0%), rectal swabs (58.3%), and cerebrospinal fluid (60.0%). Blood-vessels in the deep cutis or subcutis were affected and the gangrenous process extended superficially. Noma in older children and adults is caused by fusospirochaetosis but noma neonatorum appears to be due to P. aeruginosa septicaemia.


Endogenous opioid peptides in neurons of the human brain. Immunoreactive leucine-encephalin has been found in nerve terminals and cell bodies of neurons in the human brain. The most intense reaction was found in the globus pallidus, followed by the spinal nucleus of the trigeminal nerve. These observations suggest that in man endogenous opioid substances may play an important part in neurotransmission in areas of the central nervous system related to pain and locomotor activity.


Dopamine and schizophrenia. A therapy revisited and revised. Evidence is presented for a modulatory role for dopamine in the C.N.S. and for an indirect rather than a direct involvement of dopaminergic systems in the production of schizophrenic symptoms. The fine structure of dopaminergic fibres indicates that a majority of presynaptic boutons are not in close juxtaposition with postsynaptic neuronal membranes. This points to a wider distribution of receptors than localisation to dense postsynaptic membrane. Preliminary studies of receptor localisation support this conclusion, providing indirect evidence for autoreceptors and evidence for localisation, in the glia, of a substantial number of receptors.


Adipose tissue, a neglected factor in aetiology of breast cancer? The abundant adipose tissue that intimately invests both the alveolar and the ductal epithelium of the mammary gland may function as a slow-release depot for lipid-soluble carcinogenic agents of both endogenous and exogenous origin taken up from the bloodstream.


Potential contribution of primary-school teachers to the health of a developing country. Like many other developing countries, Kenya gives higher priority to primary education than to health services. Thus, in rural areas primary schools are evenly distributed, well attended, and staffed by well-qualified teachers whereas health centres are thinly scattered, inaccessible to many, and staffed largely by people of low educational attainment. The public-health knowledge of school teachers exceeds that of many health workers. It is suggested that teachers of health science, in particular, represent a valuable resource for preventive and promotive medicine which can be exploited at little cost.