The Merkel cell system and a comparison between it and the neurosecretory or APUD cell system. The Merkel cell, a neural crest migrant to the skin, possesses a characteristic intranuclear rodlet, cytoplasmic membrane-bound granules, and horn projections and is usually associated with nerve terminations. It is also associated with all types of known organizations of sensory nerve endings in the skin and possesses characteristic enzyme reactions in relation to the ending. The Merkel cell is found in the skin and the oral mucous membranes, and similar cells are observed in the taste buds and pulmonary tissue. A comparison between the Merkel cell and the APUD cell system of polypeptide hormone and amine-producing cells reveals many structural and chemical similarities.


The influence of oestradiol on the metabolism of androgens by human prostatic tissue. The uptake and metabolism of testosterone, androstenedione and 5alpha-dihydrotestosterone (DHT) by human benign hyperplastic prostates and prostatic carcinomas have been measured in organ culture. DHT was a major metabolite of both testosterone and androstenedione in the benign tissue and the androstanediols were the principal metabolites of DHT. Over half the carcinomas produced less DHT from testosterone than the benign hyperplastic prostates, and carcinomas from the oldest patients showed an enhancement of 17beta-hydroxysteroid dehydrogenation. There was no relationship between these differences in metabolism and the degree of differentiation of the carcinomas. Oestradiol decreased the production of DHT from both testosterone and androstenedione and, at low androgen concentrations, increased the production of androstanediols from testosterone, androstenedione and DHT. Uptake of DHT, but not of the other two androgens, was stimulated by oestradiol.


Biochemical investigations of separated epithelium and stroma from benign hyperplastic prostatic tissue. Using arginase and hydroxyproline as biochemical markers, the yields and homogeneity of separated epithelial and stromal tissues from surgically removed benign hyperplastic prostate glands have been assessed. On the basis of these markers, about 30% of epithelial and 95% of stromal tissues were recovered. Dehydroepiandrosterone sulphate sulphatase activity was found predominantly in the epithelium, whereas testosterone 5alpha-reductase activity was predominantly in the stroma.


Carrier-directed anti-hapten responses by B-cell subsets. The capacity of the trinitrophenyl (TNP) haptenic group, coupled to a series of chemically dissimilar carriers, to cross-stimulate putative T- dependent and T-independent murine B-cell subpepulations was determined by using an in vitro limiting dilution technique to generate primary IgM responses. It was found that TNP-Ficoll and TNP-dextran, two T- independent antigens with little or no polyclonal mitogenicity, stimulate the same population of anti-TNP precursors, which is distinct from the precursor population activated by TNP-bacterial lipopolysaccharide (LPS), a T-independent polyclonal mitogen, or TNP-horse erythrocytes (HRBC), a T-dependent antigen. On the other hand, TNP-LPS and TNP-HRBC activate the same precursor population, indicating that LPS can substitute for the T- cell signal in T-dependent B-cell responses, whereas nonmitogenic T- independent antigens cannot. However, the cumulative evidence from this and other laboratories strongly indicates that LPS and T-dependent antigens activate B cells by different mechanisms. Of particular interest, LPS is incapable of activating B cells responsive to weakly- or nonmitogenic T-independent antigens. Based on clonal burst size, T-dependent antigens are capable of inducing greater antigen-specific B-cell proliferation than T-independent antigens. However, TNP conjugates of Ficoll and dextran, which are relatively poor inducers of polyclonal B-cell activation, induced larger anti-TNP clones than did TNP-LPS, a strong polyclonal mitogen. The findings reinforce the evidence favoring existence of multiple B- cell subpopulations with distinctive activation pathways. They also strengthen the proposition that a given B-cell subset can be activated by more than one mechanism.


B-cell tolerance. III. Effect of papain-mediated cleavage of cell surface IgD on tolerance susceptibility of murine B cells. Under defined conditions, papain removes IgD from cells while leaving IgM, H-2, Ia, Lyb-2, and complement receptor intact. The effect of such treatment with papain on the induction of tolerance in murine splenic B cells was determined in an in vitro system. Treatment of the cells with papain has no effect on subsequent antibody responsiveness presumably because surface receptors regenerate before and during incubation with immunogen. Removal of increasing amounts of IgD results in increasing susceptibility of thymus-dependent responsive cells to tolerance induction. The tolerance susceptibility of thymus-independent responsive cells, which we have previously suggested are immature cells that bear only IgM, is unaffected by cleavage of IgD. If cells are incubated for 24 h after treatment with papain, cell surface IgD and tolerance resistance return. These results indicate that a surface molecule affects susceptibility of B cells to induction of tolerance and suggest that this molecule may be IgD.


Properties of reticulum cell sarcomas in SJL/J mice. V. Nature of reticulum cell sarcoma surface antigen which induces proliferation of normal SJL/J T cells. The results of studies on the reticulum cell sarcoma (RCS) tumors of SJL/J mice presented here, indicate that spontaneous tumors, which arise in older mice, also possess the capacity to induce the vigorous proliferative response in syngenetic T lymphocytes that are characteristic of the transplantable RCS lines. Analysis of cell surface antigens revealed the presence of Ia determinats on gradient-purified transplantable RCS tumor cells; however, these cells did not express Thy 1.2, nIg, or, any of the viral proteins that were tested for by specific antisera. Pretreatment of RCS cells with anti-Ia sera and complement-deleted cells that were stimulatory for syngenetic T lymphocytes, and addition of anti-Ia sera directly to cultures blocked the proliferative response at the stimulator (RCS) cell level. Lymph node cells from H-2(8) strains other than SJL/J, including A.SW and B10.S also gave proliferative responses to RCS cells, although lower in magnitude. A requirement on the part of responding cells for identity with RCS cells at the Ir region was indicated by the finding that A.TH but not A.TL lymph node cells responded to RCS. It is concluded that RCS cells stimulate Ir-region identical T cells (without evidence of presensitization) through a modification in the expression of Ia antigens on the surface of the tumor cells.


Human histocompatibility determinants and virus antigens: effect of measles virus infection on HLA expression. Histocompatibility antigens on the surface of human lymphoblastoid cells were quantified by a microadsorption technique. During the course of measles virus infection, no quantitative or qualitations in surface HLA antigens were observed. In contrast, infection with poliovirus type 1 or vesicular stomatitis virus, or treatment with puromycin (50 microgram/ml) resulted in a significant decrease in surface HLA. These experiments suggest that an inhibition of host protein synthesis rather than the insertion of virus-specificied antigens into the membrane results in a net decrease in amounts of this cell surface antigen. The HLA antigens also appear to be both functionally and structurally distinct from measles virus surface antigens. Pretreatment of cells with HLA-directed antibody did not prevent the infection of these cells by measles virus, thus HLA antigens appear unrelated to the measles virus receptor site on the plasma membrane. Electron microscopic studies revealed that measles virus maturation occurs at membrane sites devoid of demonstrable HLA. Furthermore, HLA antigens could not be detected on the surfaces of mature infectious virions.


B-lymphocytes activation by the Fc region of IgG. Strong stimulation of DNA synthesis (up to 150-fold) and blast transformation can be induced in mouse spleen cells by Fc fragments of human IgG. The mitogenic response is optimal on day 5 of culture and is dependent on the concentration of Fc fragments with a sedimentation rate of 3-5S. Intact IgG is also stimulatory, but only when modified by heat aggregation, and produces only a 10-fold increase in [3H]thymidine uptake. The stimulation by aggregated IgG is dependent on the Fc portion, since aggregated (or soluble) Fab or F(ab')2 fragments are inactive. The results show that the response is T-cell independent and that it is a function of nylon wool adherent, surface Ig-positive, Fc receptor-bearing B lymphocytes. Fc fragments do not induce plaque-forming cells to human IgG in normal mouse spleen cell cultures, but rather trigger polyclonal antibody synthesis (anti-goat erythrocytes, anti-2,4,6-trinitrophenyl). It is postulated that the Fc region of antibodies plays a role in the regulation of the humoral immune response by triggering clonal expansion of B lymphocytes.


Immunosuppressive factor(s) specific for L-glutamic acid50-L-tyrosine50 (GT) II. Presence of I-J determinants on the GT-suppressive factor. The responses to the synthetic antigens, L-glutamic acid(60)-L- alanine(30)-L-tyrosine(10) (GAT) and L-glutamic acid(50)-L-tyrosine(50) (GT) are controlled by genes in the I region of the mouse H-2 complex (1-3). Preimmunization of the mice bearing the H-2(p,q,s) nonresponder haplotypes with GAT stimulates the development of suppressor T cells that inhibit in vivo or in vitro antibody responses to GAT complexed to the immunogenic carrier, methylated bovine serum albumin (GAT-MBSA) (4). The copolymer GT is not immunogenic in any inbred mouse strain tested, and has a suppressive effect on the antibody responses to GT-MBSA in mouse strains bearing the H-2(d,f,k,s) haplotypes; suppressor T cells have been demonstrated to be responsible for specific GT suppression (3). We have obtained specific suppressive extracts from thymus and spleen cells of GAT-or GT-primed suppressor strains (5,6). The specific suppressive T-cell factors in the active extracts have been characterized (6,7) and appear similar to the carrier-specific suppressor factor described by Tada and Taniguchi (8). These products belong to a family of newly identified molecules coded for by the I region of the H-2 complex with affinity for antigen and helper (9,10) or suppressive (5-8) regulatory activity on the immune response. Recently, Tada et al. have reported that the keyhole limpet hemocyanin (KLH)-specific suppressor factor is coded for by the I-J subregion of the H-2 complex (11). We now demonstrate also that a GT-specific suppressor factor extracted from the spleens and thymuses of B10.BR (H-2(k)) mice bears determinants controlled by the I-J subregion of the H-2 complex.


The I-J subregion codes for determinats on suppressor factor(s) which limit the contact sensitivity response to picryl chloride. The cell-mediated immune reactivity (CMI) of mice to contact chemicals such as picryl chloride (PCI) is influenced by thymus-derived suppressor T lymphocytes (1,2). The development of these suppressor T lymphocytes is stimulated by the intravenous administration of 2,4,6-trinitrobenzene sulfonic acid (TNBS). Zembala and Asherson have further demonstrated that a specific suppressor factor(s) can be detected in the supernates of cultured suppressor T cells. This factor suppresses the transfer of contact sensitivity (CS) to PCl (1,2). In experiments reported elsewhere (3), we have shown that the PCl suppressor supernates of Zembala and Asherson can also suppress the development of contact sensitivity to PCl. The immunochemical analysis of suppressor factor (SF) operative in the CS response to PCl has revealed many similar properties (3) to other suppressive moieties functioning to limit the plaque-forming cell (PFC) response to dinitrophenylated-keyhole limpet hemocyanin (DNP-KLH) as well as the strict antigen specificity of each respective suppressive factor, suggested that there might be a common origin of these substances. Indeed, in each case these respective factors were found to bear determinants controlled by the H-2 gene complex (4,5). Recently, in selected systems, the I-J subregion has been found to code for the Ia determinants present on suppressor cells (6) and suppressor factors (4,5). In accord with these findings, we report that antigen-specific SF which limit the CS response to PCl bear I-J determinants, implying that analogous suppressive regulatory mechanisms in CMI as well as antibody responses may be determined by genes of one subregion of the H-2 complex.


Induction of immunological tolerance requires that the B cells can respond to the polyclonal B-cell-activating properties of the thymus-independent antigens. Mice were rendered specifically tolerant to the fluorescein isothiocyanatedextran (FITC) epitope by injection of FITC-dextran B512. Their spleen cells were removed at various times and cultivated in vitro with different polyclonal B-cell activators, such as lipopolysaccharide (LPS), purified protein derivative of tuberculin, and native dextran. LPS caused the appearance of high affinity anti-FITC plaque-forming cells to an equal extent with cells from untreated and tolerant animals, whereas native dextran failed to activate cells from tolerant mice, although it was a potent activator of normal cells. It was concluded that tolerance induction only affects those B cells that could respond to the polyclonal B-cell-activating properties of the tolerogen, but not other B cells having an identical set of Ig receptors directed against the tolerogen.


Regulatory functions of hapten-reactive helper and suppressor T lymphocytes. I. Detection and characterization of hapten-reactive suppressor T-cell activity in mice immunized with hapten-isologous protein conjugate. Helper and suppressor T-cell activities were detected simultaneously in the spleen cells of mice immunized with para-azobenzoate (PAB)-mouse gammaglobulin (MGG). Dinitrophenyl (DNP)-specific B cells were raised by immunization with DNP-keyhole limpet hemocyanin (KLH) and used as the indicator B-cell population. The helper and suppressor T-cell activities were determined after adoptively transferring spleen cells from PAB-MGG- primed donors and DNP-KLH-primed donors into X-irradiated recipients. Stimulation of these recipients with DNP-MGG-PAB detected helper T-cell activity, which was measured in terms of increased anti-DNP antibody responses of DNP-KLH-primed cells over these responses in the presence of unprimed cells. On the other hand, when DNP-KLH-primed cells were stimulated with DNP-KLH-PAB in the presence of PAB-MGG-primed cells, anti-DNP antibody responses were substantially lower than in unprimed normal cells. This suppressor cell population was (a) hapten-reactive, (b) present in B-cell-depleted spleen cells, (c) Thy-1 positive, (d) detectable earlier than the helper T-cell activities after priming (e) more radiosensitive than helper cells, and (f) found in the spleen but not the lymph nodes in contrast to helper T cells. These data indicate that these suppressor T cells are distinct from the helper T cells. PAB-reactive T cells clearly suppressed the antibody response by inhibiting KLH-reactive helper T-cell functions. The hapten-reactive T-lymphocyte system described here should be useful for analyzing and manipulating the immune response and for studying regulatory interactions of helper and suppressor T cells in the induction of antibody responses.


Regulatory functions of hapten-reactive helper and suppressor T lymphocytes. II. Selective inactivation of hapten-reactive suppressor T cells by hapten-nonimmunogenic copolymers of D-amino acids, and its application to the study of suppressor T-cell effect on helper T-cell development. An experimental condition was established in vivo for selectively eliminating hapten-reactive suppressor T-cell activity generated in mice primed with a para-azobenzoate (PAB)-mouse gamma globulin (MGG)-conjugate and treated with PAB-nonimmunogenic copolymer of D-amino acids (D- glutamic acid and D-lysine; D-GL). The elimination of suppressor T-cell activity with PAB-D-GL treatment from the mixed populations of hapten- reactive suppressor and helper T cells substantially increased apparent helper T-cell activity. Moreover, the inhibition of PAB-reactive suppressor T-cell generation by the pretreatment with PAB-D-GL before the PAB-MGG-priming increased the development of PAB-reactive helper T-cell activity. The analysis of hapten-specificity of helper T cells revealed that the reactivity of helper cells developed in the absence of suppressor T cells was more specific for primed PAB-determinants and their cross-reactivities to structurally related determinants such as meta-azobenzoate (MAB) significantly decreased, as compared with the helper T-cell population developed in the presence of suppressor T lymphocytes. In addition, those helper T cells generated in the absence of suppressor T cells were highly susceptible to tolerogenesis by PAB-D- GL. Similarly, the elimination of suppressor T lymphocytes also enhanced helper T-cell activity in a polyclonal fashion in the T-T cell interactions between benzylpenicilloyl (BPO)-reactive T cells and PAB- reactive T cells after immunization of mice with BPO-MGG-PAB. Thus inhibition of BPO-reactive suppressor T-cell development by the BPO-v-GL- pretreatment resulted in augmented generation of PAB-reactive helper T cells with higher susceptibility of tolerogenesis to PAB-D-GL. Thus, these results support the notion that suppressor T cells eventually suppress helper T-cell activity and indicate that the function of suppressor T cells related to helper T-cell development is to inhibit the increase in the specificity and apparent affinity of helper T cells in the primary immune response. The hapten-reactive suppressor and helper T lymphocytes are considered as a model system of T cells that regulate the immune response, and the potential applicability of this system to manipulating various T cell-mediated immune responses is discussed in this context.


Molecular internalization of a region of myelin basic protein. The conformation of myelin encephalitogenic or basic protein (BP) was investigated with a double-antibody radioimmunoassay by studying the reaction of BP or its fragments with antibodies produced in two rabbits against peptide 43-88 linked to rabbit serum albumin. Both antisera reacted well with peptide 43-88 but showed little or no reaction with BP. Absorption of these antisera with a BP-immunoadsorbent did not remove the antibody activity against peptide 43-88. Within the region of peptide 43- 88 it was shown that peptides 68-88 and 79-88 gave an equivalent or better reaction than peptide 43-88, whereas peptides 43-67 and 64-73 had very little reactivity. In the BP fragments containing region 43-88, peptide 1-88 showed the best reactivity, peptide 20-166 showed minimal reactivity, while peptide 1-115 showed none. These data document the internal position of at least a portion of peptide 43-88 and all of residues 79-88 in the BP molecule. The much greater reactivity of peptide 1-88 as compared to peptide 1-115 suggests that the region or a portion of the region of BP containing residues 89- 115 participates in the conformational alignment of BP restricting access to peptide 79-88. After absorption with BP, neither of the antisera prepared to peptide 43-88 reacted with PNS myelin in fixed tissue sections but continued to react with CNS myelin in similarly treated sections. The present findings demonstrate the need to consider the role of shielded antigenic determinants in the investigation of antigens or of immune responses.


Direct visualization of T lymphocytes bearing Ia antigens controlled by the I-J subregion. Previous studies have demonstrated that I-J-subregion-controlled Ia antigens are only expressed on a small subpopulation of peripheral T lymphocytes which includes the suppressor T cells of antibody responses (6). This subpopulation of T cells cannot be detected by conventional dye-exclusion cytotoxicity tests. A sensitive rosetting procedure therefore was developed for detecting the binding of anti-Ia antibodies to T lymphocytes. This assay system, unlike the complement lysis technique, has a low background and since it represents a direct binding assay could detect noncomplement-fixing antibodies in the antisera. Anti-Ia sera were absorbed with B cells and using the rosetting procedure in genetic mapping studies the remaining antibodies were found to be directed against I-J-subregion-controlled determinants. These determinants were shown to be highly haplotype specific for H-2(k) and H-2(s) and appeared to be exclusively expressed on Ly-l.l(-), Ly2.1(+), T lymphocytes, at least some of which were suppressor T cells. Lymphoid organs differed in their content of anti-I-J-reactive cells, the hierarchy being spleen, lymph node more than thymus, bone marrow. In contrast, on a T-cell basis, a high proportion (35 percent) of the T cells in bone marrow reacted with anti-I-J antibodies, a substantial proportion (13 percent) of T cells from spleen were reactive, whereas the lymph node and thymus T-cell populations contained only a small proportion of positive cells (1-4 percent).


A suppressor T cell in the human mixed lymphocyte reaction. Lymphocytes from an HLA-B7 DW2 homozygous multiparous woman, J.H., failed to respond in the mixed lymphocyte reaction to lymphocytes from her DW1 homozygous husband, W.H., and certain other homozygous typing cells. J.H. lymphocytes could suppress the response of HLA matched responders to W.H. This effect was shown to be radiosensitive and due to a T cell. The suppressor cell showed antigen specificity.


Sharing of Ia antigens between species. I. Detection of Ia specificities shared by rats and mice. A mouse anti-rat xenogeneic antiserum, B10.D2 anti-BN, has been found to react with a subpopulation of lymphoid cells of certain mouse strains. The corresponding alloantiserum, B10.D2 anti-B10.BR, reacted in analogous fashion with lymphoid cells of BN rats. In the case of the cross-reaction on mouse cells, mapping studies indicated that at least part of the reactivity was with the product of gene(s) determined by the I-A subregion of the H-2 complex. Chemical isolation studies with radiolabeled cell surface preparations indicated that the antigens detected in both mouse and rat had mol wt characteristic of Ia antigens (35,000 and 28,000 dalton molecules). Testing of fractionated spleen cell populations revealed that the cross-reactive antigens were expressed predominatly on B cells, but that a subpopulation of T cells were also reactive. Wider strain and species distribution studies are in progress to determine the extent of such Ia cross-reactions between species and to further assess the practical and theoretical importance of such cross-reactions.


The specific antigen-binding cell populations of individual fetal mouse spleens: repertoire composition, size, and genetic control. In order to analyze the genetic and physiological basis of controls affecting the generation of the repertoire of antigen-binding cells in fetal mice, we have measured the numbers of spleen cells specific for each of four antigens as a function of the total numbers of nucleated and Ig-bearing cells in inbred, hybrid, and random bred fetuses. For each of the two inbred strains BALB/c and CBA/J, the proportion of nucleated cells specific for a given antigen was the same for all individuals of the strain at the 18th day of gestation. The proportion did vary from antigen to antigen, however, and for each antigen the proportion of specific cells observed in CBA/J fetuses was approximately four times that observed in BALB/c fetuses. This difference appeared to be due to a difference between the two strains in the relative size of the repertoire of antigen-binding spleen cells at this stage of development, inasmuch as the frequency of Ig-bearing spleen cells in CBA/J fetuses was likewise approximately four times that observed in BALB/c fetuses. In random bred Swiss-L fetal mice at the 18th day of gestation, the proportion of cells specific for a given antigen varied significantly from one individual to the next. The ratio of proportions of the two antigens observed was constant from individual to individual, however, and this constant ratio differed significantly from the ratio observed for the same two antigens in fetal BALB/c and CBA/J inbred mice. These data suggest that the ontogeny of the repertoire of antigen-binding cells in fetal mice is subject to at least two independent sets of controls, one affecting the relative size of the repertoire in the spleen, and the other affecting the distribution of antigen-binding specificities within that repertoire. Analysis of repertoire size and composition in the spleens of hybrid fetuses confirmed the observation that the two parameters are controlled independently, and suggested further that the control of repertoire size in these fetuses is due to the action of one or a few closely-linked autosomal Mendelian genes. These data are consistent with models for the origin of antibody diversity in which the genes coding for the full repertoire of antibodies are generated somatically from a small number of germ-line genes early in development and in the absence of any strong positive or negative selection with respect to antigenic specificity.


Evolution of an idiotypic determinant: anti-Val. An antibody population which reacts only with human sickle cell hemoglobin (HbS) and not with normal human hemoglobin, has been isolated from goat, sheep, and guinea pig antisera. These antibody populations termed anti-Val (Val), isolated from an individual goat (no. 6) and sheep (no. 26) have been used to elicit anti-idiotypic responses in rabbits. These anti-idiotypic sera were used to study the idiotypic cross-reactions between the goat and sheep anti-Val. Strong cross-reactions were present using either Ra anti-goat anti-Val or Ra anti-sheep anti-Val. Guinea pig anti-Val did not cross-react with these anti-idiotypic sera. Binding of HbS to the anti-Val of the goat and sheep could be blocked by the anti-idiotypic sera, but the binding of HbS to the guinea pig anti-Val could not. These data demonstrate idiotypic cross-reactivity between two closely related species.


Enrichment of antigen-specific B lymphocytes by the direct removal of B cells not bearing specificity for the antigen. Antigen-specific B cells (ASC) were purified from other B cells by prior incubation with specific antigen followed by rosetting with erythrocytes conjugated with anti-mouse Ig and sedimenting on Ficoll-Isopaque. This procedure allowed the removal of most of the B cells, while those speicifc for the antigen used in incubation were retained. Relative to the B-cell content, ASC were enriched 64- to 132-fold. The method is highly specific in that B cells primed to two different antigens, turkey gamma globulin and sheep erythrocytes, could be separated from each other. The advantages of this indirect purification procedure over purification procedures which obtain ASC directly are the simplicity of obtaining the ASC and the ability of the ASC of respond to antigen without the addition of other cells.


In vitro generation of tumor-specific cytotoxic lymphocytes. Secondary allogeneic mixed tumor lymphocyte culture of normal murine spleen cells. In vivo or in vitro immunity to murine leukemia virus (MuLV)-induced leukemia cells which do not effectively produce virus, has been difficult to demonstrate. Because immunizations with allogeneic murine leukemia cells have been used to confer syngeneic tumor immunity to virus- producing cells, we attempted to generate lymphocytes, cytotoxic to syngeneic nonproducer leukemia cells, by stimulating normal murine spleen cells with allogeneic nonproducer leukemia cells in mixed tumor lymphocyte culture (MTLC) reactions in vitro. Secondary allogeneic MTLC of normal C57BL/6 or DBA/2 spleen cells effectively produced syngeneic tumor-specific cytotoxic lymphocytes. Target cells lysed in lymphocyte- mediated cytolysis (LMC) assays, included both Friend and Rauscher virus- induced syngeneic murine leukemia cells and chemically-induced hematopoietic tumor cells. Syngeneic tumor cells were lysed regardless of whether they produced infectious MuLV or expressed viral antigens gp-71, p-30, or p-12 at the cell surface. Syngeneic normal cells (thymus, lymph node, or Concanavalin A-stimulated spleen cells) used as targets in LMC assays were uneffected by lymphocytes harvested from secondary allogeneic MTLC. Several other in vitro culture treatments including secondary syngeneic MTLC and repetitive mixed lymphocyte culture stimulations were incapable of generating tumor-specific cytotoxic lymphocytes. Based upon these results, we propose that secondary MTLC stimulation of normal spleen cells with allogeneic nonproducer leukemia cells selects for the proliferation of two subpopulations of antigen-specific cytotoxic lymphocytes. The population capable of effecting syngeneic tumor cell lysis is directed against tumor-associated cell surface antigens which may be distinct from viral structural proteins or glycoproteins. The growth of these tumor-specific cytotoxic lymphocytes may be enhanced by a soluble allogeneic effect factor produced by the proliferation of the second subpopulation of lymphocytes generated in repetitive allogeneic MTLC, namely those lymphocytes with specificities directed against differing histocompatibility antigens.


The guinea pig I region. I. A structural and genetic analysis. The Ia antigens of the guinea pig have been shown to play a central role in the regulation of the immune response. We have previously partially characterized the chemical structure of these antigens. In this communication, we further characterize the structure of the five Ia antigens already described, as well as two new Ia antigens. Evidence is presented which shows that these seven Ia antigens can be organized into three distinct groups, each with a characteristic structure. The Ia.2 determinant of strain 2 and the Ia.3 and Ia.5 determinants of strain 13 animals are found on molecules composed of a 25,000 dalton chain and a 33,000 dalton chain in noncovalent association, or else are individually expressed on nonlinked 33,000 and 25,000 dalton molecules. The Ia.4 and Ia.5 determinants of strain 2 and the Ia.7 determinant of strain 13 are borne on 58,000 dalton molecules in which two chains are linked by disulfide bonds. The Ia.1 and Ia.6 determinants of strain 13 are found on a molecule of 26,000-27,000 daltons. Ia.6 of strain 2 has yet to be definitively assigned. Furthermore, in strain 13 animals the Ia.3 and Ia.5 determinants are borne on the same molecule, as are the Ia.1 and Ia.6 determinants. In strain 2 animals, the Ia.4 and Ia.5 determinants are found on the same molecule. On the basis of chemical structure, we have divided the guinea pig I region into three subregions. The accompanying paper presents evidence of associations between particular Ia genes and Ir genes.


The guinea pig I region. II. Functional analysis. We have examined whether an association exists between specific Ia antigen genes and Ir genes which are encoded within the same haplotype. Functionally monospecific sera to the Ia antigens of the guinea pig MHC were selective in their ability to inhibit antigen-specific T-cell proliferation and we were thus able to demonstrate an association between individual Ia specificities and specific Ir genes. The results of these studies in inbred animals were confirmed by examining the association of Ir genes and Ia antigens in the outbred guinea pig population. Of great interest was the observation that antisera made against cross-reactive Ia antigens of strains lacking specific Ir genes would still inhibit immune responses of strains possessing the Ir gene, if the Ir gene was associated with that Ia antigen in the responder strain.


Cytotoxic T cells distinguish between trinitrophenyl- and dinitrophenyl-modified syngeneic cells. Spleen cells sensitized against trinitrophenyl (TNP)-modified stimulator cells displayed a cytotoxic effect against syngeneic TNP-modified but not dinitrophenyl (DNP)-modified target cells. The same finding was observed in the opposite direction; that is, effector cells sensitized against DNP-modified stimulator cells did not cross kill TNP-modified targets. The specificity of the anti-TNP effector cells was confirmed in a cold target competition assay. Presensitization in vivo with hapten-modified cells followed by rechallenge and testing in vitro did not alter the specificity of the response between the haptens. These data indicate that the receptor(s) on the cytotoxic T cell can distinguish between two closely related haptenic molecules.


Selective response to H-Y antigen by F1 female mice sensitized to F1 male cells. T-cell mediated cytotoxic responses to H-Y antigen require co-recognition of H-Y and H-2 gene products. F1 mael stimulating cells and target cells express H-Y antigen in association with both parental H-2 haplotypes. However, F1 females primed in vivo and challenged in vitro with F1 male cells lyse male target cells of F1 and only one parental H-2 haplotype. Thus, (CBA X B10)F1 females sensitized to (CBA X B10)F1 male cells lyse (CBA X B10)F1 and CBA but not B10 male target cells, and (BALB/c X B10)F1 females sensitized to (BALB/c X B10)F1 male cells will lyse (BALB/c X B10)F1 and B10 but not BALB/c male target cells. It is suggested that this may represent an effect of immune response or suppressor genes mapping in the major histocompatibility gene complex which regulate responsiveness to H-Y antigen.


Induction of virus-specific modifications recognized by cytotoxic T cells is not altered by prior substitution of target cells with trinitrophenol. Cytotoxic thymus-derived lymphocytes generated after interaction with trinitrophenyl (TNP)-substituted or virus-infected cells only lyse H-2 compatible target cells modified with the component used to immunize (TNP or virus). Prior saturation of TNP-reactive sites inhibits neither the infectivity of influenza A viruses, nor the capacity of infected cells to develop antigenic changes recognized by influenza-immune T cells. The two antigens are distinct entities on the cell membrane and do not obviously compete to form interactions with H-2 molecules.


Cytotoxic T lymphocytes induced against allogeneic I-region determinants react with Ia molecules on trinitrophenyl-conjugated syngeneic target cells. The major histocompatibility complex codes for determinants which are recognized by and serve as targets for cytolytic T lymphocytes (CTL) (1). Antigens coded for by the K and D loci of the H-2 complex can activate xenogeneic or allogeneic CTL (2,3). In addition, the H-2K or H-2D gene products function as those molecules against which syngeneic CTL responses specific for chemical, viral, and minor H antigens are directed (4-8). It has recently been shown that Ia determinants can also serve as target antigens for distinct but weaker CTL responses (9-13). Those clones which recognize Ia antigens see them independently of K- or D- coded antigens as shown in genetic studies and by antisera-blocking experiments (12,13). We have proposed that the existence of clones of CTL specific for I-region-coded determinants is not fortuitous; rather these clones specifically recognize Ia determinants and may have an immunoregulatory role. These CTL may affect those immune functions which are at least partially dependent on or controlled by I-region-coded molecules. Two predictions can be made and tested concerning the role of Ia determinants in cytolytic systems and the role, if any, of I-region- specific CTL in regulating the immune response: (a) that if as we and others have shown, certain Ia specificities can serve as a third series of major histocompatibility antigens, then Ia antigens should be susceptible to the same types of antigenic modifications as H-2K- or H-2D-coded structures and thus serve as targets for CTL directed against modified-self in selected systems; and (b) that allogeneically induced I-region-specific CTL should demonstrate cross-reactivity with targets bearing modified syngeneic I-region-coded determinants. Data will be present which demonstrates that trinitrophenyl (TNP)-modified syngeneic I-region determinants can serve as targets for CTL induced by allogeneic Ia antigens.


Drug risks in pregnancy revisited.

Drug risks in pregnancy revisited. In the important interface of assuring safe care for the pregnant mother and her developing child that falls to the family physician, current knowledge of drug risks in pregnancy is essential. A recent survey showed that an average of 4.5 drugs are taken throughout the pregnancy. Eighty percent are self-prescribed, and a positive correlation was shown between the incidence of congenital anomalies and drug intake. Numerous recent reports of previously unrecognized fetal complications of commonly prescribed and self-used drugs and heightened public awareness makes it imperative that family physicians know the risk principles of major drug groups and be able to recognize drug-induced symptoms and signs in the older infant as well as in the infant.


Distinction between envelope antigens of murine xenotropic and ecotropic type C viruses by immunoelectron microscopy. The indirect ferritin-labelled antibody technique was used to determine the reactivity of an antiserum prepared against the NZB xenotropic virus with three murine xenotropic viruses, a feline xenotropic virus and a murine ecotropic virus. The envelope antigens of the xenotropic type C viruses isolated from the NZB, NIH Swiss and C57L mice were tagged with ferritin. The feline RD114 virus was not. Gross murine leukaemia virus was tagged, but only at high serum concentrations. The cross-reactivity titre of Gross virus to anti-NZB serum was removed by a serum dilution which was still reactive to xenotropic viruses. This difference in reactivity titres between a xenotropic and an ecotropic virus was sufficient to distinguish one from the other in doubly infected cultures. Specific tagging of membrances of cells infected by xenotropic virus was also observed.


Biological properties of human leukocyte interferon components. Human leukocyte interferon, purified approximately 1000-fold by affinity chromatography on immobilized anti-interferon globulins and SDS-Sephadex filtration, was resolved into one major and one minor component by adsorption chromatography on hydroxylapatite and electrophoresis in polyacrylamide gels. These components were indistinguishable in their capacity to protect bovine, porcine and murine cells, and the antiviral activities of both were equally susceptible to reduction by beta-mercaptoethanol. They were neutralized to the same degree of rabbit anti-leukocyte interferon but were not neutralized by rabbit antifibroblast interferon serum. Mice immunized with either component developed antibodies to both but failed to form antibodies against human fibroblast interferon. Our present evidence indicates that the two components posses at most only minor structural and antigenic dissimilarities.


A histofluorescence study of events accompanying accumulation and migration of norepinephrine within locally cooled nerves. Glyoxylic acid was used to induce fluorescence in sections of rabbit sciatic nerve. In fresh nerves treated with this agent there were scattered finely beaded axons with a weak blue-green fluorescence. During local cooling, blue-green fluorescence accumulated steadily at the proximal boundary of the cooled region but never at its distal boundary. This accumulation gave rise to dilated axons that often swelled into brilliantly fluorescent balloon-like structures up to 10 microgram in diameter. Axonal fluorescence was probably specific for norepinephrine, being enhanced by inhibition of the metabolism and diminished by inhibition of the synthesis or storage of this neurotransmitter. After local cooling of nerves for 1.5 hr, specific fluorescence was confined within 0.8 mm of the cooled region. Rewarming led to rapid removal of fluorescence from the cooled region and to disappearance of most of the balloon-like swellings. Simultaneously, rewarming caused brightly fluorescent fibers that were neither dilated nor swollen to appear in distal regions of nerve. As this wave of fluorescence migrated distally with increasing duration of rewarming, it was spread over increasingly broad regions of nerve, which suggests that axonal transport of norepinephrine may invole some kind of dispersive process.


Focal chemotherapy of brain tumours using semipermeable membranes. Semipermeable silastic rubber membranes can be used to diffuse focally a variety of chemicals including antitumour drugs. Of the eight drugs tested, in vitro tests showed the best diffusion with dactinomycin, mithramycin, and oncovin, and the poorest diffusion with bleomycin, fluorouracil, and thiotepa. Biological testing was performed with mithramycin, dactinomycin, and oncovin using tissue cultures of human glioblastoma and subcutaneous implants of mouse ependymoblastoma. All three drugs caused rapid tissue culture cell death with direct injection, and impeded tumour growth when given intraperitoneally. Dactinomycin by silastic diffusion proved more effective than mithramycin against tissue cultures, but neither drug had a significant effect against local tumours treated with implanted drug capsules. Silastic diffusion of oncovin reduced tumour sizes significantly ipsilateral to the implant, compared to contralateral implants and to untreated controls.


Multiple sclerosis: sensitization of a myelin basic protein fragment (peptide T) encephalitogenic to primates. A preliminary report. Myelin basic A1 protein is the sole antigen of the central nervous system capable of inducing experimental allergic encephalitis (EAE), but sensitization with peptide fragments of the molecule may also induce disease. Using the macrophage migration inhibition factor (MIF) assay we have compared sensitization to portions of the molecule active in inducing EAE in monkeys with results obtained concomitantly using the intact protein. Cellular sensitization to human myelin A1 protein, peptide L (residues 1-116), peptide T (residues 117-170), and petide Y (residues 154-170) was studied using the Thor-Rocklin MIF assay system. Lymphocytes of 10 normal subjects, 10 multiple sclerosis patients 0-3 weeks after onset, 10 4 weeks to 3 months after and 10 6 months or longer after onset of an acute exacerbation were assayed. Results of the investigation reveal evidence of cellular sensitization to myelin basic protein encephalitogenic peptide T occurring during attacks of multiple sclerosis. Peptide L, relatively nonencephalitogenic to primates, failed to induce a significant lymphocyte response, whereas peptide Y which is encephalitogenic gave irregular results.


Calcifying epithelial odontogenic tumor: histochemical properties of homogeneous acellular substances in the tumor. Calcifying epithelial odontogenic tumor (CEOT) is characterized by the presence of homogeneous, acellular, eosinophilic, and calcifying substances. The histochemical properties of homogeneous acellular substances are compared to enamel matrix in the human developing teeth. Homogeneous amorphous material in the CEOT is not amyloid histochemically; the composition of materials are compared to the proteins of enamel matrix. The findings of CEOT from the Japanese literature are introduced clinically and radiographically.


Vascular changes in the brain of spontaneously hypertensive rats: hyaline and fibrinoid degeneration. The arterial changes designated as "hyalinosis" in the vessels of spontaneously hypertensive rats were studied using light- and electron-microscopy. Such changes were classified into three types, hyaline, fibrinoid and atypical fibrinoid degeneration. Hyaline material consisted mainly of excessive lamellar thickening of the basement membrane synthesised by endothelial and smooth muscle cells and associated with cellular debris and electron-dense particles. Acid mucopolysaccharides were found to be one of the components of the hyaline material. The nature of fibrinoid material was primarily fibrin derived from polymerised fibrinogen which has permeated through the injured endothelial cell layer. Atypical fibrinoid material consisted of granular and fibrillar substances derived from blood plasma. In some cases, all three changes were encountered in the same artery.


Alpha1-fetoprotein in neonatal hepatobiliary disease. It has been suggested that the quantitative estimation of serum alpha-1-fetoprotein may help in distinguishing the neonatal hepatitis syndrome from biliary atresia. We measured the serum AFP concentration in 52 neonates and infants with various hepatobiliary disorders, including neonatal hepatitis syndrome (group I), biliary atresia (group II), and other hepatopathies such as choledochal cyst (group III). The mean serum AFP concentration in patients with neonatal hepatitis was significantly greater than the mean concentration in the other two groups. There was no significant difference between the mean serum AFP concentrations in patients with biliary atresia and in group III patients. Patient age was noted to be an important factor: Serum AFP levels greater than 35 microgram/ml in infants one to four months of age suggpst the diagnosis of neonatal hepatitis syndrome. Serum AFP levels below 10 microgram/ml in infants less than four months of age suggest the diagnosis of biliary atresia or hepatopathies other than neonatal hepatitis. However, the variable and significant overlapping of serum AFP values between 10 and 35 microgram/ml limit the diagnostic value of this test.


Antithyroid drug therapy for toxic diffuse goiter (Graves disease): thirty years experience in children and adolescents. Since 1946 104 children and adolescents with toxic diffuse goiter have been treated. Sixty-seven were treated with a thioamide for more than 12 months. Remission occurred in 61%. Twenty-five patients had a 12-hour perchlorate discharge test to determine the least frequent dose schedule required for disease control: 68% could be controlled on a single daily dose and an additional 25% on an every 12-hour schedule. A one-hour radioiodine uptake on combined therapy and change in thyroid gland size during treatment were found to be highly correlated with the presence of a spontaneous remission. Thirty-six patients have been treated by subtotal thyroidectomy. To date 65% have developed permanent hypothyroidism. In the author's opinion, thoamides are the treatment of choice for the majority of children and adolescents.


Antimicrobial action of compound 48/80 against protozoa, bacteria, and fungi. Compound 48/80 inhibited the growth of protozoa, bacteria, and fungi but had no effect on the multiplication of viruses. All susceptible organisms were inhibited by 10 microgram/ml of crude compound 48/80, and some were inhibited by as little as 0.1 microgram/ml. Against Tetrahymena pyriformis, this drug was seven times more potent than quinine. Separation of compound 48/80 into different fractions indicated that some antimicrobial activity could be separated from the histamine-liberating activity. It was found that compound 48/80 is not surface active at 500 microgram/ml.


In search of anti-Trypanosoma cruzi drugs: new leads from a mouse model. Nine of 25 carefully selected compounds (from a stock of more than 200 000 chemical species amassed principally as a result of testing against other parasitic diseases) were found to have significant suppressive activity against the parasites in the blood of a Trypanosoma cruzi mouse model. Eight of these compounds evaluated in this model had suppressive activity equal to or greater than the reference compound, nifurtimox. For the first time, suppressive activity against T. cruzi is reported for a 7-aminoquinoline, a phosphonium salt, and TAC pamoate; The biological model is believed to be able to serve as a means of identifying other new "leads* in seeking drugs broadly effective against T=ruzi infections in man.


Conformational analogues of antihypertensive agents related to guanethidine. In an effort to clarify the conformational requirements, if any, of agents producing adrenergic neuronal blockade through mechanisms similar to guanethidine, the synthesis and pharmacological evaluation of 15 analogues of cinnamylguanidine are described. These analogues represent derivatives in which the distance between the center of the ring system and the guanidinium nitrogen atom varies from 3.9 to 6.2 A. While conformational relationships could not be defined in this study, three analogues (3, 4, and 5) were apparently more potent than guanethidine in the in vitro assay employed.


Diastereoisomeric N-tetrahydrofurfurylnoroxymorphones with opioid agonist--antagonist properties. The two diastereoisomeric N-tetrahydrofurfurylnoroxymorphones and their hydrochlorides 1a and 1b have been prepared and studied pharmacologically; The N-(R)-tetrahydrofurfuryl derivative 1a proved to be an opioid agonist--antagonist and the N-(S)-tetrahydrofurfuryl derivative 1b a pure antagonist. As an analgesic, 1a is 25 times more potent than morphine, but it does not show morphine-like side effects in mice. In withdrawn morphine-dependent rhesus monkeys, 1a only partially suppresses abstinence. Its therapeutic ratio is exceptionally favorable compared with those of morphine and pentazocine. As antagonists, 1a and 1b have comparable potencies of 0.25 and 0.20 of that of nalorphine, respectively, in vivo. In vitro, however, 1a is 28 times (guinea pig ileum) or 6.5 times (mouse vas deferens) more potent than 1b. The antagonist properties of 1a and 1b were not anticipated according to known structure--activity relationships.


Chemotherapy of murine ovarian carcinoma by methotrexate-antibody conjugates. Antibodies with specificity for an experimental ovarian carcinoma were coupled to methotrexate by two procedures. Water-soluble carbodiimide gave effective coupling, but a large proportion of the conjugate was rendered insoluble, presumably due to alteration or aggregation of the antibodies. A modification of the mixed anhydride procedure gave rise to products which were completely soluble and allowed a high degree of coupling to be achieved. In vivo testing of the conjugates revealed a significant increase in survival time in treated mice when compared to a variety of control groups; these included groups receiving antiserum or drug alone, mixtures of the two, and conjugates of normal gamma-globulin with methotrexate. Our results provide added support for the concept that tumor-associated antibodies coupled to cytotoxic agents are more effective than single agents or noncoupled mixtures of agents.


Staining reticulin in beef dorsal lymph nodes for electron microscopy. Reticulin has been classically identified by virtue of its argyrophilia, as observed by light microscopy. The similiarity between reticulin and collagen has been a matter for considerable discussion. This paper describes the application of a silver-staining procedure which demonstrates the fine structure of these fibres when studied by electron microscopy, giving a direct indication of the morphological identity of the argyrophilic structures, i.e. reticulin, with collagen. Beef lymph node tissue was used for this study.


Effect of specific immunotherapy with preimmunization against alpha-fetoprotein on a mouse transplantable hepatoma. Antibodies to autologous alpha-fetoprotein (AFP) were produced in mice by immunization with rat AFP. C57L/J mice with or without such antibodies were inoculated sc or ip with controlled numbers of cells of the syngeneic, AFP-producing, BW 7756 hepatoma. There was a linear relationship between circulating AFP and tumor mass, with elevated AFP being detectable earlier than the tumor. The AFP levels of the immunized mice were generally lower than those of control mice, and tumors could be detected before elevated concentrations of AFP appeared in the circulation. An extensive series of transplantations with varying protocols for immunization did not protect against tumor and did not affect the rate of tumor growth.


Rapid grain and cell counting for cell kinetic studies. A commercially available bacterial colony counter has been adapted for the counting of radioautographic grains over individual cells in smears and for the counting of cells in histologic sections. For radioautographic grains, the correlation coefficients between counts obtained visually by 2 observers and between counts obtained visually and with the use of the instrument were similar (r=0.999 and r=0.998, respectively). The instrument counts were obtained more rapidly than the visual counts and were associated with less observer fatigue. Even though the performance of the instrument in counting cells in mouse bone marrow sections was less accurate than that in counting radioautographic grains, a good estimation of marrow cell number was obtained (r=0.968). Data on bone marrow cellularity were obtained far more rapidly than those with semiquantitative methods.


Effect of busulfan on oncornavirus-like activity in platelets and chromosomes in polycythemia vera and essential thrombocythemia. The effect of busulfan therapy on the activity of oncornavirus-like particles and chromosome patterns in patients with polycythemia vera and essential thrombocythemia was studied. Three patients had pretreatment platelet counts greater than 1 million/microliter, abnormal bone marrow karyotypes, and electron microscopic and biochemical evidence of oncornavirus. The results demonstrated that in all 3 patients virus-like particles and reverse transcriptase-like activity could no longer be found in the platelets within 2-4 weeks after the initiation of busulfan treatment. The platelet count was still elevated at this point for each patient, although the count returned to normal levels within 2-3 weeks after virus-like activity was no longer detectable. The chromosome patterns, characterized by aneuploidy (30-50%) before treatment, improved after therapy.


Specificities in human cell-mediated cytotoxicity. Specificity of natural cell-mediated cytotoxicity was investigated through selective reactions detected by direct cell-mediated cytotoxicity and by inhibition of cytotoxicity through competition. Assuming that target cells reacting alike in direct cytotoxicity shared common antigens, we classified 10 target cells into three groups by target antigens: TA (target antigen) 1, 2, and 3. Partial confirmation of the three groups was achieved in the cross-competition assay. The distinction of TA 1 as a group was clear but some cross-reactivity existed between TA 2 and TA 3 cells.


Relationship between leukemia antigens and stimulation in mixed leukocyte culture. Leukemia cells from 13 of 27 patients stimulated lymphocytes of their HLA-identical siblings in mixed leukocyte cultures (MLC). Stimulation correlated with high background uptake of tritiated thymidine and high percentages of cells that stained with fluorescein-conjugated polyvalent goat antiserum against human immunoglobulin. Results were similar to those reported with fractionated autologous lymphocyte subpopulations in normal individuals. Investigation of leukemia-specific stimulation in MLC is therefore problematic.


Antigenic characterization of flavivirus structural proteins separated by isoelectric focusing. Isoelectrofocusing of nonionic-detergent-disrupted flaviviruses separated the envelope glycoprotein of 53,000 to 58,000 daltons and the nucleocapsid protein of 14,000 daltons. The envelope protein and nucleocapsid protein were isolated at isoelectric points of pI 7.8 and 10.3, respectively. The antigenic determinants of St. Louis encephalitis, Japanese encephalitis, and dengue virus envelope and nucleocapsid proteins were examined by solid-phase competition radioimmunoassay. By the appropriate selection of antiserum and competing proteins, it was possible to distinguish type-specific, complex-reactive and flavivirus group-reactive antigenic determinants. The envelope glycoproteins of St. Louis encephalitis, Japanese encephalitis, and dengue viruses were found to contain each of these three classes of antigenic determinants. Most of the determinants on the envelope protein were type specific, some were complex reactive, and a small fraction were flavivirus group reactive. The nucleocapsid protein contained only flavivirus group-reactive antigenic determinants.


Lactate dehydrogenase isoenzymes in human prostatic fluid: an aid in recognition of malignancy? A total of 1,435 specimens of prostatic fluid were collected from 1,076 patients by rectal massage. Lactate dehydrogenase isoenzymes in the fluid were separated by acrylamide disk electrophoresis and the ratio of lactic dehydrogenase-5/lactic dehydrogenase-1 was used as the parameter for the test. There were significant differences in the ratio among patients with a normal prostate, benign prostatic hyperplasia and prostatic carcinoma. The mean values of lactic dehydrogenase-5/lactic dehydrogenase-1 ratios were 0.48 +/- 0.09, 1.36 +/- 0.17 and 5.21 +/- 0.79, respectively. Of 30 patients with histological evidence of prostatic carcinoma 25 (85 per cent) had a ratio greater than 2 in the prostatic fluid. On the other hand the ratio was less than 2 in 50 of 57 men (88 per cent) with a  histological diagnosis of benign prostatic hyperplasia devoid of inflammation. When prostatic fluid contained white blood cells a positive correlation was observed between the ratio and the number of white blood cells per microscopic field (r equals 0.289, p less than 0.001). The results indicate that a lactic dehydrogenase-5/lactic dehydrogenase-1 ratio exceeding 2 in the prostatic fluid in the absence of inflammation should be regarded as an indication of high risk of the presence of carcinoma of the prostate.


Biochemical and immunological characterization of the major envelope glycoprotein gp69/71 and degradation fragments from Rauscher leukemia virus. Analysis of the proteins of Rauscher murine oncornavirus by immunoprecipitation showed that antiserum to the purified envelope glycoprotein of approximately 69,000 and 71,000 daltons (gp69/71) reacted as well with a number of other components of several murine oncornaviruses of approximately 45,000, 32,000, and 15,000 daltons. Polypeptides of similar size were also produced by limited proteolysis of purified gp69/71; these degradation fragments were shown to contain carbohydrate by the incorporation of (3)H from sodium boro[(3)H]hydride after neuraminidase and galactose oxidase treatment. Each of these glycoproteins was isolated by preparative polyacrylamide gel electrophoresis and was analyzed by tryptic peptide mapping. The major virion components of 69,000 and 71,000 daltons were nearly identical, as were the primary degradation fragments. Analysis of the immunological properties of the glycoproteins showed that the 71,000-, 69,000-, and 32,000-dalton glycoproteins behaved similarly with respect to type and group-specific antigenic determinants. In contrast, the 45,000-dalton glycoprotein lacked detectable interspecies and some of the group-specific reactivity. Components of about 45,000 and 32,000 daltons isolated directly from virions were also identified as constituents of the major envelope glycoprotein by immune precipitation and tryptic peptide mapping. These results indicate that all of the examined virion glycoproteins of approximately 71,000, 69,000, 45,000, and 32,000 daltons are derived from the same viral gene and that these lower-molecular-weight glycoproteins can readily be produced from the major envelope glycoprotein.


Intranuclear microtubules in lung mast cells of guinea pigs in anaphylactic shock. Intranuclear microtubules were found in lung mast cells during anaphylactic shock following ovalbumin challenge of guinea pigs passively sensitized with homocytotropic antibodies. Simultaneously, such cells showed mitochondrial swelling with a clear matrix and partially disrupted cristae. Although a decreased number of mast cell granules generally accompanied these observations, the degree of degranulation and the morphologic appearance of the granules are less reliable criteria to indicate the immediate participation of a cell in the anaphylactic phenomenon. The cells that presented intranuclear microtubules were mainly found in the vicinity of the bronchiolar smooth muscle. The distribution of cations has been investigated in these tissues with a combined oxalate-pyroantimonate method. Whereas in controls the reaction product is located mainly in the nucleus and the mitochondria of mast cells, these sites become almost completely devoid of precipitate during the anaphylactic reaction. A hypothetical link between histamine release, intracellular distribution of cations, possibly calcium, and the appearance of intranuclear microtubules in mast cells is proposed.


Maternal serum-alpha-fetoprotein measurement in antenatal screening for anencephaly and spina bifida in early pregnancy. Report of U.K. collaborative study on alpha-fetoprotein in relation to neural-tube defects. Nineteen centres collaborated in a study to determine the efficiency of maternal serum-alpha-fetoprotein (A.F.P.) measurement as a method of screening for neural-tube defects (N.T.D.S.) between 10 and 24 weeks of pregnancy. Data were collected on 18 684 singleton pregnancies and 163 twin pregnancies without fetal N.T.D.s, and on 301 singleton pregnancies with fetal N.T.D.s (146 with anencephaly, 142 with spina bifida, and 13 with encephalocele). The best time for detecting open spina bifida by measuring maternal serum-A.F.P. is at 16-18 weeks of pregnancy. In clinical practice, serum-A.F.P. cut-off levels expressed as multiples of the normal median may be more convenient to use than percentiles because they are easier to derive and more stable. Also, the proportion of affected pregnancies with serum-A.F.P. levels exceeding a given multiple of the median is unlikely to vary significantly from centre to centre or over time. In contrast, the proportion of unaffected pregnancies with A.F.P. levels exceeding a given multiple of the normal median will vary depending on the precision with which serum-A.F.P. and gestation are measured. At 16-18 weeks of pregnancy 88% of cases of anencephaly, 79% of cases of open spina bifida, and 3% of unaffected singleton pregnancies had A.F.P. levels equal to or greater than 2-5 times the median for unaffected singleton pregnancies. At this gestation age the numbers of unaffected pregnancies with A.F.P. levels above 2-5 times the normal median can be reduced by about a third if women with borderline A.F.P. levels are retested, although this would not greatly change the detection-rate of affected pregnancies. In the United Kingdom as a whole, women with serum-A.F.P. levels above 2-5 times the normal median at 16-18 weeks of gestation will have an approximately 1-in-20 chance of having a fetus with open spina bifida; the risk of having any N.T.D. will be approximately 1 in 10. The results of this study indicate that screening pregnant women by measuring the concentration of A.F.P. in their serum is an effective method of selecting women for ultrasonography and amniocentesis so that N.T.D.s can be diagnosed in utero.


Evaluation of a sucrose/electrolyte solution for oral rehydration in acute infantile diarrhoea. An oral sucrose/electrolyte solution brought about complete hydration in 19 out of 20 consecutive children with a median age of 1 year with moderate to severe dehydration due to acute diarrhoea (1 child did not respond and needed intravenous therapy). Vomiting, abdominal distension, and appearance of sugar in the stools during oral hydration did not interfere with its success. A satisfactory response was shown by weight gain (mean +/- S.E. = 9-2 +/- 0-6%), restoration of plasma-bicarbonate to normal levels, falls in the haematocrit values and in the plasma specific gravity, and complete clinical recovery. Disaccharidases in jejunal mucosal biopsy specimens in the acute phase showed reduced lactase in 75% and reduced sucrase in 18%; maltase levels were adequate. Except for the child who did not respond, all showed a satisfactory rise of blood-sugar after sucrose and glucose tolerance tests. Sucrose is cheap and easily obtained; its use instead of glucose in electrolyte solutions would be a considerable gain.


Merozoite vaccination of douroucouli monkeys against falciparum malaria. Erythrocytic merozoites of Plasmodium falciparum (Gambia) were isolated from cultures of schizont-infected human red cells on CF 11 cellulose columns. Douroucouli monkeys vaccinated with such preparations stored in liquid nitrogen and then emulsified in Freund's complete adjuvant (F.C.A.), were resistant to successive challenges with West African (Lagos) and East African (Uganda Palto-Alto) strains of P. falciparum. The induced immunity is specific since vaccination with P. knowlesi merozoites in F.C.A. does not modify the course of P. falciparum infections in douroucouli monkeys.


Immunisation with a polyvalent pneumococcal vaccine. Reduction of adult respiratory mortality in a New Guinea Highlands community. A double-blind controlled trial of a 14-valent pneumococcal polysaccharide vaccine was carried out in 11 958 adults at Tari in the Papua New Guinea Highlands. Pneumococcal infection, confirmed by blood-culture and lung aspirate, was less in the vaccinated group by 84%. Mortality from pneumonia was less by 44%.


Comparison of oral 25-hydroxycholecalciferol, vitamin D, and ultraviolet light as determinants of circulating 25-hydroxyvitamin D. Circulating concentrations of 25-hydroxyvitamin D (25-OHD) were measured during short-term and long-term oral treatment with 25-hydroxycholecalciferol (25-OHD3), 25-H.C.C.) or with vitamin D in over 200 subjects over a period of 5 years. Ten times more vitamin D than 25-OHD3 was required to produce equivalent plasma-25-OHD concentrations. Plasma-25-OHD was a power function of dosage with both compounds. These data indirectly measure the superior therapeutic potency of 25-OHD3, show that dose-response relations with both compounds may be useful in diagnosis, and indicate that there are pronounced constraints on 25-hydroxylation of vitamin D. Together with the effects of ultraviolet light, now shown to be equivalent to oral vitamin D in doses of 8000-10 000 I.U. daily, these constraints may protect against vitamin-D deficiency in winter.


Prophylactic antimicrobial drug therapy at five London teaching hospitals. A report by the Study Group on the Use of Antimicrobial Drugs. We report some findings of a survey of antimicrobial prophylaxis in 5 London teaching hospitals. It is practised predominantly in surgical patients, both before and after operations, penicillins being by far the commonest drugs. Striking differences were found, particularly in the duration of treatment in different hospitals in the same specialty, and also between consultants in the same hospital. The findings raise important questions that need to be answered by clinical trials.


Relation between smoking and age of natural menopause. Report from the Boston Collaborative Drug Surveillance Program, Boston University Medical Center. The relation of cigarette smoking to natural menopause has been evaluated in two independent sets of data. In both, at each age from 44 to 53 years, more current smokers were post-menopausal compared to "never" smokers. In the first set of data the age-standardised proportions of women who were postmenopausal were 35, 36, 43, and 49%, respectively, for never smokers, ex-smokers, and current smokers of 1/2 and greater than or equal to 1 pack per day. The corresponding proportions for the second set were 53, 59, 60 and 65%.


Treatment of fulminant hepatic failure by polyacrylonitrile-membrane haemodialysis. 24 patients with fulminant hepatic failure who had deteriorated to grade-IV coma were treated by repeated periods of haemodialysis with a polyacrylonitrile membrane. 9 patients fully recovered consciousness, and 8 (33%) survived to leave hospitals. These results are to be compared with those of conservative management alone (15% survival in 53 cases) and those obtained initially with charcoal haemo-perfusion (38%). Of the 16 treatment failures, cerebral oedema was found at necropsy in 13 (18%). Whether this would have been less of problem if treatment had been started earlier in the course of the illness remains to be determined.


Altered plasma-lipids associated with oral contraceptive or oestrogen consumption. The Lipid Research Clinic Program. Mean plasma cholesterol and triglyceride concentrations were measured in White female users and non-users of oral contraceptives and oestrogens in 10 diverse, demographically defined North American populations. About 50% of the younger women (20-24 years old) were taking oral contraceptives. In these women mean triglyceride concentrations were up to 48% higher and mean cholesterol concentrations were about 5% higher than in non-users. The 95% percentile of the total lipid distribution among non-users was used to define hyperlipidaemia. In young women on oral contraceptives hypercholesterolaemia was up to three times more common and hypertriglyceridaemia was up to five times more common than in non-users. 37% of older women (50-54 years) (presumably intramenopausal and postmenopausal) were hormone users, and in this group there were small, inconsistent alterations in plasma-triglyceride and a modest but consistent reduction in mean cholesterol concentration.


Predictive classification of human breast carcinomas based on lactalbumin synthesis. The milk protein lactalbumin was found in the cytosol fraction of 20 of 53 human breast carcinomas. Oestrogen receptor (E.R) was detected in 36 of the tumours. The presence of lactalbumin was closely associated with the presence of E.R.; in tumours containing both, there was a positive linear correlation between concentrations of lactalbumin and E.R. Synthesis of lactalbumin by a mammary carcinoma seems to indicate an intact oestrogen receptor mechanism and may therefore be useful in identifying tumours responsive to endocrine therapy.


Patterns of sexually transmitted enteric diseases in a city. In the past three years cases of shigellosis, amoebiasis, and viral hepatitis A and B have increased fourfold to tenfold in San Francisco. These diseases were most common in people who had adopted "alternative life-styles", especially in young men, In many cases there was a history of frequent orogenital and oral-anal sexual contact between men with no common source. Despite energetic public-health measures and intensive efforts by physicians treating these cases, the increase has not yet been brought under control. Other cities may experience similar difficulties with sexual transmission of enteric diseases.20


Prophylactic systemic antibiotics in colorectal surgery. The prophylactic value of gentamicin combined with either lincomycin or metronidazole in 52 patients undergoing colorectal surgery was investigated. The results confirmed the value of this practice. In a control group, the sepsis-rate was 48% with 1 death attributable to sepsis, compared with a sepsis-rate of 4% in the treated group. The combination of gentamicin and lincomycin was effective against sepsis but pseudomembranous colitis developed in 2 of the 14 patients treated with this combination of drugs. Lincomycin was discontinued, and when metronidazole was substituted the results were equally good and there were no toxic side-effects.


Intravenous glucose, aminoacids, and fat in the postoperative period. A controlled evaluation of each substrate. The metabolic effects of hypocaloric (2-51 MJ), equicalorific quantities of intravenous glucose, crystalline aminoacids, and soyabean emulsion were comapred with those of fasting in 4 groups of male patients (closely matched for age, weight, and fat-free mass) on the day of vagotomy and pylorplasty and on the fast 3 postoperative day. Patients given glucose alone excreted less nitrogen than the fasting patients but nitrogen sparing was greatest in the group given aminoacids alone. There was no evidence of any nitrogen sparing when intravenous fat was the source of energy. There was no difference in blood glucose, free fatty acids, and insulin concentrations among the groups. Ketone-body concentrations rose in the fasting and fat-fed groups but remained low in the groups who received glucose or aminoacids. This study indicates that, in the short term, the increased preservation of protein achieved by the infusion of aminoacids compared with that produced by the traditional infusion of glucose does not warrant the extra cost involved.


Difficult labour as a cause of communicating syringomyelia. Although some causes of communicating syringomyelia are known most cases of the disease can only be described as idiopathic. The results of a questionnaire suggest a high incidence of difficult labour in the mothers of syringomyelia patients. A high proportion of patients had forceps deliveries, and a high proportion were the first born in their families. Because the validity of taking a birth history at the age of presentation (mean age 40 y, S.D. 14 for this sample) might be questioned, patients admitted with other disorders (mean age 40 y, S.D. 16) were used as a control group. These patients matched the syringomyelia patients in social class and place of birth. It seems likely that birth trauma may be a cause of tonsillar descent through the foramen magnum (ectopia) and of arachnoiditis; both conditions are often present. Once the tonsils become engaged in the foramen magnum, difference between the cranial and spinal pressure may, over the course of several years, cause the tonsils to descend further, thus leading to communicating syringomyelia. Over half these patients have a history of difficult labour and it is possible that birth injury may be a factor even where birth is regarded as normal.


Diagnosis of hyperthyroidism when serum-thyroxine alone is raised. 31 patients admitted to hospital with different non-thyroidal intercurrent diseases were found to have raised total serum-thyroxine (T4) and free T4 together with normal serum-triiodothyronine (T3) concentrations. At admission none was clinically hyperthyroid. Patients were assigned to 3 groups according to clinical course and the laboratory findings. In the first group (14 patients) classic hyperthyroidism developed after recovery from the intercurrent disease. 11 out of these 14 patients had recently received iodine-containing preparations. In a second group (11 patients) the initially raised serum-T4 rapidly returned to normal with recovery from the non-thyroidal disease. In a third group (6 patients) serum-T4 concentrations remained raised well after recovery from intercurrent disease. In this group, there were 2 cases of transient iodine-induced (Jod-Basedow) hyperthyroidism in which raised serum-T4 returned spontaneously to normal after several months as iodine was eliminated. These results indicate that increase in serum-T4 with normal serum-T3 in patients with intercurrent systemic disease is not always the result of hyperthyroidism and in many cases probably reflects changes in peripheral metabolism of T4. It is suggested that careful clinical follow-up is needed in patients with raised serum-T4 and normal serum-T3 for the early detection and treatment of classic hyperthyroidism.


Effect of insulin antibodies on free and total plasma-insulin. Insulin administration often causes the development of insulin antibodies. Plasma insulin-binding capacity was assessed and free and total plasma insulin were measured in 96 insulin-dependent diabetic patients. Patients with the highest insulin binding capacity not only had the highest total insulin but also had the lowest free insulin. It is concluded that bound insulin is not unavailable to cells, that sufficient dissociation can occur for this insulin source to make a sizeable contribution to insulin delivery to the cell receptors, and that this can happen at a very low free insulin concentration.


Contacts between young patients with Hodgkin's disease. A case-control study. To investigate the possibility that Hodgkin's disease (H.D.) may be transmitted from person to person, a case-control study was conducted among 87 of the 97 H.D. patients diagnosed under the age of 40 years in the period 1962-71, resident at the time of diagnosis in a defined area around Oxford. For each of the 87 H.D. patients a matched control patient was selected with a diagnosis other than that of a chronic or malignant disease. H.D. patients and controls were interviewed to determine their places of schooling and work and attempts were made to link pairs of patients who had attended the same school or work place at the same time. The findings do not support the hypothesis that H.D. patients may pass on the disease to others. Among the H.D. patients, links between 40 pairs were established, compared with 40-75 links expected, based upon the experience of all 174 persons studied. The only finding which was statistically significant (p approximately 0-04) arose when H.D. patients were considered to be "susceptible" from 10 to 5 years before diagnosis and "infective" from diagnosis to 2 years after diagnosis: 7 pairs of links were found against 2-75 expected. However, since 7 postulated periods of susceptibility and 9 postulated periods of infectivity were examined, this one "statistically significant" finding might easily have arisen by chance.


Acute pontine syndromes following head injury. Patients in the early stage of severe head injuries occasionally present with a clinical picture analogous to that in vascular or neoplastic pontine lesions and they can be identified by careful clinical examination and by repeated electroencephalography (E.E.G.). The commonest features in these cases were preservation of consciousness, constriction of the pupils, disturbances of reflex ocular motility, severe pyramidal deficits, decerebrate rigidity, myoclonic manifestations, irregular breathing, and cranial nerve palsies. An E.E.G. tracing similar to the normal pattern was a common finding. These observations suggest that pontine syndromes may be directly produceg a non-reacting, but conscious, patient for one who is deeply comatose.


Subclinical ergotism.

Subclinical ergotism. The systolic blood-pressure at the ankle and the first toe was measured in 30 patients, mean age 42, who had taken ergotamine regularly for more than a year. With one exception, the patients had no symptoms or signs of arterial insufficiency in the limbs, but all had low-normal or abnormal foot systolic blood-pressures. In a group of 13 patients who stopped taking ergotamine the distal pressures rose significantly and from the ninth day were normal.


Diagnostic importance of changes in circulating concentrations of immunoreactive trypsin. A specific and sensitive radioimmunoassay (R.I.A.) has been developed which makes possible the determination of serum or plasma trypsin concentrations despite the presence of trypsin inhibitors, which have invaldiated previously available enzymatic techniques. The assay was most precise at about 300 microng trypsin standard Ag5 per litre of serum, a value comparable with the mean in 76 healthy volunteers (273 microng/1) and in 20 hospital patients with non-pancreatic disease (266 microng/1). Markedly raised concentrations (970-6500 microng/1) were found in all 14 patients with acute pancreatitis and in 8 patients with chronic renal failure (580-1360 microng/1). Abnormal concentrations were found in 11 of 16 patients (69%) with pancreatic cancer (8 high, 3 low) and in 15 of 23 patients (65%) with chronic pancreatitis (3 high, 12 low). Patients with jaundice had normal or marginally lower than normal concentrations unless pancreatic disease or common-duct gallstones were present.


Lecithin consumption raises serum-free-choline levels. Consumption of choline by rats sequentially increases serum-choline, brain-choline, and brain-acetylcholine concentrations. In man consumption of choline increases in levels in the serum and cerebrospinal fluid; its administration is an effective way of treating tardive dyskinesia. We found that oral lecithin is considerably more effective in raising human serum-choline levels than an equivalent quantity of choline chloride. 30 minutes after ingestion of choline chloride (2-3 g free base), serum-choline levels rose by 86% and returned to normal values within 4 hours; 1 hour after lecithin ingestion, these levels rose by 265% and remained significantly raised for 12 hours. Lecithin may therefore be the method of choice for accelerating acetylcholine synthesis by increasing the availability of choline, its precursor in the blood.


Leukaemia in benzene workers.

Leukaemia in benzene workers. Workers occupationally exposed to benzene in 1940-49 were followed for vital status up to 1975. In comparison with two control populations, a significant (P less than 0-002) excess of leukaemia was observed. A five-old excessive risk of all leukaemias and a ten-fold excess of deaths from myeloid and monocytic leukaemias combined are demonstrated in the study population compared with controls. These figures under-estimate the true leukaemia risk to benzene-exposed workers, because follow-up is only 75% complete and the untraced 25% of the study population were all regarded, in the statistical analysis, as being alive at the end of the study period. The environment of the workers in the study population was not contaminated with solvents other than benzene, and existing records indicate that the benzene levels themselves were generally below the limits recommended at the time of their measurement.


Radiotherapy and hyperbaric oxygen in head and neck cancer. Final report of first controlled clinical trial. We report the results of a prospective controlled trial of the effect of hyperbaric oxygen as an adjuvant in radiotherapy of head and neck cancer. Patients were allocated randomly to treatment in oxygen or air. The radiotherapy in both groups was identical in planning, dose, and fractionation--i.e., 3500 rads in 10 fractions in 3 weeks. There was no difference in the survival rate between the two groups. However, significantly better local tumour control was seen in the hyperbaric-oxygen group, particularly in smaller lesions; there was significantly greater need for salvage surgery in the air group. Radiation effects on normal tissue appeared somewhat greater in the oxygen series, especially on laryngeal cartilage.


Radiotherapy and hyperbaric oxygen in head and neck cancer. Interim report of second clinical trial. A controlled clinical trial is in progress to assess the value of hyperbaric oxygen and radiotherapy in the management of head and neck cancer. An established dose-fractionation schedule in hyperbaric oxygen is being compared with a widely used conventional schedule in air. Survival and local recurrence-free rates are significantly higher in the oxygen group, and the effects on normal tissue are similar in both groups. These findings suggest a genuine therapeutic advantage. There was a distinct improvement in the results of treating advanced laryngeal carcinoma, where there was a high survival rate, without resort to laryngectomy.


25-hydroxy-vitamin-D in nephrotic syndrome. Serum-25-hydroxy-vitamin-D (25-OHD) nephrotic syndrome (N.S.) without renal insufficiency (urinary protein excretion greater than 3-5 g/24 h/1-73 m2; glomerular filtration-rate greater than 80 ml/min/1-73 m2). Serum-25-OHD levels were low in patients with N.S. (mean 19 nmol/1, range 4-41 nmol/1), compared with a normal range of 25-200 nmol/1. Serum-concentrations of Gc-globulin--the binding protein for vitamin D and its metabolites (D.B.P.)--were significantly (P less than 0-001) lower in patients with N.S. (mean 340 mg/1, range 190-480 mg/1) than in non-proteinuric controls (mean 440 mg/1, range 376-510 mg/1, measured by radial immunodiffusion). In contrast to non-proteinuric urine, urine of all N.S. patients contained a large amount of 25-OHD-binding capacity; D.B.P. could be detected in all N.S. urines after concentration. Scatchard analysis of the urine demonstrated the presence of a low-affinity and a high-affinity binding protein (tentatively identified as albumin and D.B.P.). These results suggest an acquired deficiency of circulating 25-OHD in N.S. secondary to urinary loss of protein-bound 25-OHD. The biological relevance of the low 25-OHD levels is unknown. There was no clinical evidence of osteomalacia (X-ray, serum-alkaline-phosphatase); however, slightly elevated serum-parathyroid-hormone (P.T.H.) levels would be compatible with borderline vitamin-D depletion.


Effect of periodic deworming on nutritional status of ascaris-infested preschool children receiving supplementary food. The weight for age of 154 undernourished preschool children in an ascaris-infested community receiving supplementary food was monitored for a year. 74 children were given tetramisole every 4 months, while 80 controls were given placebo. The prevalence of ascaris (roundworm) infection and the severity of protein-energy malnutrition were the same in the two groups at the onset of the study. Stool-positive rates for ascariasis came down significantly in the experimental group during the year, but eradication of the worm was not possible owing to constant re-exposure. Nutritional status remained unaltered in the controls but improved strikingly in the treated children 8 and 12 months after the start of the study. Periodic deworming should form a part of the supplementary feeding programme if malnutrition is associated with ascariasis in the community.


Bowel transit-time and stool weight in populations with different colon-cancer risks. It has been suggested that the risk of cancer, polyposis, and diverticulosis of the large bowel increases with bowel transit-time. Hence, Japanese inhabitants of Hawaii, in whom the risk of these diseases is high, would be expected to have longer transit-times than Japanese in Japan, in whom such risks are low. However, bowel transit-times were similar in Japanese groups. Stools from the Hawaii Japanese did weight significantly less than the specimens from Japan, and this factor may be indirectly related to the risk of colorectal cancer, polyposis, or diverticulosis in the Japanese.


Liver dysfunction in hypertension.

Liver dysfunction in hypertension. Liver-function tests measured routinely in hypertensive patients attending the Glasgow Blood Pressure Clinic were abnormal in 15-8% of men and 6-2% of women. The patients studied appeared to be representative of the whole clinic population. Liver dysfunction was related to alcohol consumption, heavy body-weight, male sex, young age, and higher diastolic blood-pressure. It is suggested that alcohol and obesity were the principal causal factors and that fatty infiltration of the liver was the probable pathology. Liver dysfunction was unrelated to treatment. Alcohol use was found to be heavy in 12% of male patients attending the clinic, and this was probably an underestimate. The possibility that alcohol abuse may have a causal role in hypertension needs further study.


Alcohol and hypertension.

Alcohol and hypertension. Liver-function data were compared in 158 unselected hypertensives and 105 normotensives aged 45-64 years. Serum concentrations of alanine and aspartate aminotransferase (S.G.O.T. and S.G.P.T.) were higher, and more often raised, in the hypertensive patients. Serum bilirubin and alkaline phosphatase concentrations were similar in both groups. In hypertensive patients aminotransferase concentrations tended to be higher in those with increased alcohol intake.


Treatment of Gaucher's disease with liposome-entrapped glucocerebroside: beta-glucosidase. An adult with Gaucher's disease was injected with liposomes containing human glucocerebroside: beta-glucosidase. After 13 months of treatment the lower border of the right hepatic lobe and moved up 3 cm, and the left lobe had shrunk. Reduction in the size of the liver was associated with relief of pressure symptoms in the left lower abdomen. Scans obtained after the injection of radiolabelled liposomes suggested improvement in function of the reticuloendothelial system.


Unilateral breast-feeding and breast cancer. Women of fishing villages in Hong Kong by custom breast-feed with only the right breast. The hypothesis that the unsuckled breast may have an altered risk of cancer development was investigated. Records of the radiotherapy divisions in Hong Kong between 1958 and 1975 were searched, and breast-cancer patients were interviewed for a detailed history of lactation. The overall left/right ratio of cancer in the breasts of 2372 women with unilateral breast carcinoma was 0-97, indicating that breast cancer was equally distributed between the two sides. Of 73 patients with a history of exclusively one-sided breast-feeding, 27 of 34 patients aged 55 or over (79-4%) and 19 of 39 patients under age 55 (48-7%) had a carcinoma in the unsuckled breast. Comparisons of patients who had nursed unilaterally with nulliparous patients and with patients who had borne children but had not breast-fed indicated a highly significant increased risk of cancer in the unsuckled breast. No statistically significant differences in laterality of breast cancer were found in 52 patients who had for convenience nursed to a greater extent from one side than the other. This study indicates that in post-menopausal women who have breast-fed unilaterally, the risk of cancer is significantly higher in the unsuckled breast and that breast-feeding may help to protect the suckled breast against cancer.


Splenectomy and subsequent mortality in veterans of the 1939-45 war. A long-term follow-up of 740 American servicemen splenectomised because of trauma during the 1939-45 war showed a significant excess mortality from pneumonia and ischaemic heart-disease. Mortality from cirrhosis was also increased, but not significantly. The findings confirm that the risk of fatal infections is increased by asplenia; however, the risk of cancer was not increased, as it is in some other immunodeficiency states. Post-splenectomy thrombocytosis and hypercoagulability may account for the increased risk of fatal myocardial ischaemia in this group.


An unusual outbreak of food-poisoning associated with meals-on-wheels. An outbreak of food-poisoning after a meals-on-wheels lunch affected 49 persons, 1 of whom died. Bacillus cereus serotype 10 and Bacillus licheniformis were isolated in large numbers from many of the patients including the deceased and from the remains of the meal. Clostridium perfringens (C. welchii) serotype 68, which was isolated from many of the patients and the deceased but not from the food, may also have been responsible. Food kept warm during distribution can produce an ideal environment for bacterial multiplication and similar outbreaks may occur again.


[The histological reaction of cialit preserved hyaline and elastic cartilage implanted to replace partially bony wall of the guinea-pig bulla (author's transl)]. Cialit preserved cartilage was used to fill defects of the bulla of porpoises and the histological healing was studied. Hyaline and elastic cartilage are showing an integration without reactions. The cartilage is resorbed consecutively by the recipient and substituted by bone. Whereas the hyaline cartilage shows a complete bony substitute and a small enlargement we can registrate that the elastic cartilage often shows a resorptive tendency with replacement of connective tissue and loss of the shape. The histological results are corresponding to the clinical results when using preserved cartilage in middle ear surgery for restoring the posterior wall.


[Bleomycin in the intratumoral treatment of advanced squamous cell carcinomas in the oto-rhino-laryngological region (author's transl)]. In 16 patients with advanced squamous cell carcinomas infiltration therapy with Bleomycin was used either peritumorally or intratumorally. In no case a chirurgical or radiological therapy could be used. The applied doses varied between 120 mg and 555 mg. With this cytostatic infiltration therapy a measurable reduction of the tumor could be achieved in all patients, in 2 patients a total and in 3 patients a partial remission could be observed. The general conditions, as well as the intake of food were improved. A further advantage was the relief of pain with this therapy.


The serological specificities of Salmonella typhi antigens. Sera prepared with two different strains of Salmonella typhi were analysed against all the soluble antigens isolated from S. typhi 0901, S. typhi Ty2 and S. typhi Vi. Agar-gel diffusion against individual sera showed that, in all the sera, antibodies were induced against somatic antigens and free proteins. Absorptions of the sera with polysaccharides, split from the somatic antigens, removed the antibodies induced against the polysaccharide and its proteinic carrier in most of the somatic antigens of S. typhi 0901. The antibodies left in the absorbed sera reacted against the proteinic moieties of more complex somatic antigens of S. typhi and against free proteins from all the analysed strains. Only the absorption with proteins removed all the precipitating antibodies from the sera. Moreover, in incomplete absorptions with proteins, the first antibodies removed are the antipolysaccharides, since antibodies are never induced against the haptenic polysaccharide but against somatic conjugates; in these the proteinic moiety eventually varies with every batch of bacteria. The sera exhausted of precipitins still agglutinate the bacteria, thus confirming the assumption that agglutinins and precipitins may be different antibodies.


Effect of various reagents on the susceptibility of target cells to the cytotoxicity of guinea pig lymphotoxin (GLT). The cytotoxic activity of guinea pig lymphotoxin (GLT) was increased when the target cells were treated with the following metabolic inhibitors: puromycin, actinomycin D, NaN3, NaF, and 2,4-dinitrophenol, whereas calf serum, N-acetyl-D-galactosamine, colchicine, and vinblastin inhibited GLT-cytotoxicity. Puromycin, an inhibitor of protein synthesis, exhibited the most potent enhancing effect on GLT-cytotoxicity. It seems likely that there are certain cellular metabolic processes depending on cellular protein synthesis which antagonize GLT-cytotoxicity. Taking advantage of this effect of puromycin, a time-saving assay method for GLT-cytotoxicity was developed.


Rapid identification and antibiotic sensitivity testing of bacteria isolated from clinical infections. A total of 834 bacterial strains isolated from urine were subjected to rapid biochemical and serological identification and rapid antimicrobial sensitivity testing using Autobac 1. For enterobacteria (742 strains) six tests (acetoin-, beta-galactosidase-, hydrogensulphide-, indole-, ornithin-decarboxylase-and urease-production) correctly identified to genus or species level more than 99% of the strains within four hours. Staphylococci and streptococci (92 strains) were identified with full accuracy within two hours using a rapid deoxyribonuclease assay and immunoelectroosmophoresis and coagglutination. The overall accuracy for the automated antibiotic susceptibility testing using Autobac was 93% as compared to the standard plate diffusion method. In terms of rapidity for 91% of the bacterial strains the susceptibility testing was completed within four hours. After five hours 99% of the strains were analysed. Our data indicate that rapid and automated assays can be accurate and furnish the physician with adequate data within 24 hours after receipt of a urinary specimen.


[Carcinoma of the head of the pancreas--diagnosis, surgical indications and results (author's transl)]. 89 cases of pancreatic or periampullary carcinoma were analysed and the following problems discussed: 1. "early" diagnosis is at present impossible--not even with E.R.C.P. or angiography, although these provide valuable information for planning the operation. 2. Indication for operation: a curative resection was possible in only 20% of cases. The apparent advantages of total pancreatectomy as against the conventional Whipple operation (increased radicality and reduced post-op. complications) have not yet paid off in terms of improved long-term survival. 3. in 46 pancreato-duodenectomies (for carcinoma as well as complicated chronic pancreatitis) the operative and hospital mortality was 4.3%.


[Possibilities for operative therapy of carcinoma of the large intestine (author's transl)]. Tumors of the colon and rectum belong to the most frequent malignant diseases in the German Federal Republic. Only operative therapy ensures an optimal rate of success. In metastasizing and advanced local tumors, additional chemotherapy and radiotherapy should be taken into consideration. Also, preoperative radiotherapy may improve the chances of a cure in rectal carcinoma. An exact classification of the tumor is essential for a reliable prognosis. The Dukes tumor classification, still frequently used today, is inadequate for clinical application. Since the UICC has withdrawn the proposed classification for tumors of the colon and rectum, the Heidelberg proposed classification is presented, which is based on the degree of infiltration of the primary tumor and on the degree of metastatic dissemination.


[The pathogenesis of postoperative abdominal wound dehiscence. Part II: Hemostasiological checks during the course in selected patient groups (author's transl)]. By reference to clinically established risk factors, patients with an increased tendency to disturbances of wound healing were selected and investigated hemostasiologically in a prospective study. Abdominal wound dehiscence occurred postoperatively in 12 of the 28 patients selected clinically as being endangered. In all collectives the Factor XIII concentration fell postoperatively and remained low in the patients with a wound rupture, whereas it rose significantly in patients with undisturbed wound healing. Connections between hemostasis and wound healing are discussed.