Volatilization of lindance from water in soil-free and flooded soil systems. Volatilization of 14C-lindane from water in planchets and under flooded soil ecosystem was investigated. Lindane disappeared faster than parathion from planchets. More rapid loss of both insecticides occurred from water than from chloroform. Loss of lindane and parathion was related to measured losses of water by evaporation. During 5-day incubation under flooded soil conditions, disappearance of lindane was faster from open vials than from sealed vials, whereas in nonflooded soil, no volatile loss of the insecticide was evident despite water evaporation. Over 5 day incubation under flooded conditions, greater volatile loss of lindane occurred in sandy soil than in alluvial soil apparanetly due to greater adsorption to the soil colloids decreasing the insecticide concentration in the standing water on the laterite soil. Under identical conditions of water evaporation, lindane loss was directly proportional to its initial concentration in the water. These results suggest that considerable loss of soil applied pesticides can occur by volatilization from the standing water in flooded rice fields, particularly under tropical conditions.


The metabolism of lindance and its metabolites gamma-2,3,4,5,6-pentachlorocyclohexene, pentachlorobenzene, and pentachlorophenol in rats and the pathways of lindance metabolism. The metabolism of lindane and its metabolites, gamma-PCCH, PCB, and PCP, has been studied in rats. Urine, faeces, blood and important organs were analyzed for free metabolities and those bound as beta-glucuronides. TeCCOL was identified as a main metabolite by GC/MS and gamma-PCCH has been evaluated toxicologically. A degradation scheme, based on experimental results and comprising only one hypothetical metabolite is proposed and discussed.


Cytochemical differentiation of nucleic acids with a Schiff-methylene blue sequence. A method is described whereby a Feulgen type of hydrolysis of deoxyribonucleic acid is carried out on paraffin sections of routinely fixed tissues by controlled exposure of the sections to Bouin's fluid. Subsequent staining with Schiff reagent followed by methylene blue distinguishes red-to purple-stained deoxyribonucleic acid from blue-stained ribonucleic acid. This Schiff-methylene blue sequence visualizes ribonucleic acid in nucleoli and the chromidial substance of various normal and neoplastic cells and provides an assessment of their protein synthetic activity. The method has proved valuable in demonstrating normal immunocytes and immunoglobulin-forming tumor cells in pathologic specimens.


Tannic acid effect on membrane of cell surface origin in guinea pig megakaryocytes and platelets. Plasma membranes in isolated guinea pig megakaryocytes and washed platelets are poorly stained with the usual methods used to outline cell membranes. The addition of tannic acid and calcium to the initial fixative is useful to enhance electron density of all surface-derived membrane systems in these cells. The method described here shows that the increased electron denisty of membrane after fixation in the presence of tannic acid occurs both at the cell surface and along the invaginated membrane systems.


Induction of immunologic tolerance in nursing neonates by absorption of tolerogen from colostrum. Deaggregatedhuman gamma-globulin (DHGG) injected into female mice within 24 hr after delivery of a litter enters the colostrum and is absorbed intact through the intestine by nursing neonates. This absorbed HGG was present in the neonatal circulation at concentrations of 0.3 to 0.6 mg/ml of serum under the experimental conditions used. This absorption of HGG by the nursing neonate resulted in a complete, specific, tolerant state to HGG. This tolerant state was stable upon adoptive cell transfer and could not be abrogated by transfer of normal syngeneic spleen cells.


Purification and characterization of a liver-specific antigen. A liver-specific antigen (F-antigen) previously demonstrated in saline extracts of BALB/c mouse liver by double immunodiffusion was isolated and characterized. The antigen was found widely distributed among mammals but absent from avian and frog liver extracts. In immunoelectrophoresis it had an electrophoretic mobility similar to that of serum beta2-globulins, was relatively thermolabile, and was precipitated at 30 to 70% saturated ammonium sulfate concentrations. Evidence was presented that this antigen is a protein or a moiety closely associated with protein. Gel-filtration on Sephadex G-200 revealed liver-specific antigenicity in the second peak. Ion-exchange chromatography on DEAE-Sephadex A-50 revealed four peaks of which only the third one exhibited liver-specific antigenicity. This active peak contained 11 polypeptides on SDS polyacrylamide gel electrophoresis. After electrophoresis on acrylamide gel in the absence of SDS, antigenic activity was detected on one fast-moving band. Extraction of the protein band followed by SDS gel electrophoresis showed one major component of m.w. 75,000 and two major bands of m.w. 72,000 and 93,000, respectively.


Effects of C-reactive protein on platelet function. I. Inhibition of platelet aggregation and release reactions. C-reactive protein (CRP) is an acute phase reactant which shares numerous functional characteristics with the immunoglobulins. In the present study CRP was found to inhibit the aggregation of human platelets stimulated by either modified human immunoglobulin or thrombin. This effect did not involve chelation of calcium or cytotoxicity, and was overcome by larger amounts of the aggregating agents. CRP also inhibited the activation but not the activity of platelet factor 3 and the release of beta-glucuronidase. Thus, CRP can inhibit multiple platelet  reactivities. We suggest that this property of CRP may play an important role in the control of platelet responsiveness during reactions of inflammation, defense, and repair.


Immunologic cross-reactivity of antigen(s) induced by drug treatment in two leukemic sublines. Previous studies have demonstrated that new antigenic specificities, not detectable on parental cells, can be induced by in vivo treatment of murine leukemic cells with anti-neoplastic agents. The immunologic properties of leukemic cells altered by treatment with 5-(3,3-dimethyl-1-triazeno) imidazole-4-carboxamide (DIC) were investigated further. Immunologic cross-reactivity between two DIC-treated leukemic sublines has been demonstrated by cell-mediated immunity in vitro and by active or adoptive immunity in vivo. Rabbit antiserum to DIC-treated sublines absorbed with the parental cells showed residual activity against the DIC-sublines that was specifically inhibited by further absorption with DIC-cells.


Generation of cytotoxic lymphocytes in vitro: response of immune rat spleen cells to a syngeneic gross virus-induced lymphoma in mixed lymphocyte-tumor culture. Spleen cells from W/Fu rats 4 to 6 weeks after immunization with syngeneic Gross virus-induced lymphoma (C58NT)D cells usually lack detectable activity in a short-term 51Cr release assay. The results presented here demonstrate that these spleen cells retain the capacity to generate significant proliferative and cytotoxic activity upon re-exposure to mitomycin C-treated (C58NT)D cells in vitro. Optimal conditions were defined in W/Fu rats for this secondary immune response in vitro to the (C58NT)D cells. The cytotoxic response was observed to be quantitative, reproducible, and specific. Optimal generation occurred 5 days after initiation of cultures with a 30:1 responding cell:stimulating cell ratio. In vitro generated cytotoxic cells inhibit tumor growth in vivo when administered as a mixture with tumor cells.


Antigen-specific and nonspecific mediatiors of T cell/B cell cooperation. II. Two helper T cells distinguished by their antigen sensitivities. Further evidence is presented for two types of helper T cells in the mouse specific for the protein antigen, keyhole limpet hemocyanin (KLH). The first cell helps B cells respond to the trinitrophenyl hapten (TNP) coupled to KLH, is primed by relatively high doses of antigen in vivo, and yet the effector cell is stimulated by very low doses of antigen in vitro. The second cell helps B cells respond to a non-cross-reacting antigen, sheep red blood cells, presumably via production of a nonspecific factor. This cell is primed by relatively low doses of antigen in vivo, but the effector cell requires relatively high doses of antigen in vitro. Thus, the two T cell types are differently sensitive to antigen dose, both in priming and challenge. The properties of T cells responding to KLH by proliferation in vitro were also studied. These cells showed the same antigen-sensitivity in vitro, as cells producing nonspecific B cell-stimulating factors.


Antigen-specific and nonspecific mediators of T cell/B cell cooperation. III. Characterization of the nonspecific mediator(s) from different sources. T cell-containing lymphoid populations produce a nonantigen-specific mediator(s) (NSM) which can replace T cell helper function in vitro in the response of B cells to sheep red blood cells (SRBC), but not to the hapten-protein conjugate, trinitrophenyl-keyhole limpet hemocyanin, (TNP-KLH). NSM produced under three conditions: 1) stimulation of KLH-primed cells with KLH; 2) allogeneic stimulation of normal spleen cells; and 3) stimulation of normal spleen cells with Con A (but not PHA) are indistinguishable on the basis of their biologic activity and m.w., estimated as 30 to 40,000 daltons by G-200 chromatography. Production of NSM is dependent on the presence of T cells. The action of NSM on B cells responding to SRBC in the presence of 2-mercaptoethanol is unaffected by severe macrophage depletion. Extensive absorption of NSM with SRBC failed to remove its activity, confirming its nonantigen-specific nature.


Role of t lymphocytes in the humoral immune response. II. T cell-mediated regulation of antibody avidity. The effect of activated T lymphocytes (ATC) on the avidity distribution of PFC in the secondary response was studied in normal mice. The total PFC response was not significantly changed for either direct or indirect PFC by administration of ATC before secondary antigen challenge. However, marked suppression occurred of indirect PFC that secreted high avidity antibody; no suppression was seen of high avidity direct PFC. At the same time, significant stimulation was seen of relative and absolute frequencies of indirect PFC that secreted middle and low avidity antibody. These effects were dependent on Thy 1-bearing, nylon nonadherent cells which demonstrated carrier specificity. In further characterization of these effects, it was found that increasing the number of ATC transferred produced progressive loss of high avidity PFC and compensatory increase in lower avidity PFC. Moreover, in these experiments, suppression of the high avidity response was inducible with the administration of ATC 5 weeks before to 3 days after the secondary immunization. Thus, it is likely that the avidity-modifying effects are dependent on T lymphocytes which influence the late stages of B lymphocyte maturation.


Shared antigens between human malignant melanoma cells and Mycobacterium bovis (BCG). This study was undertaken to investigate the antigenic relationships between human malignant melanoma cells and Mycobacterium bovis (BCG). Rabbits were immunized with sonicates of BCG or with malignant melanoma cells from different patients and the resulting antisera were tested for their capacity to bind radiolabeled soluble extracts prepared from BCG and melanoma cells. The binding of antibodies to radiolabeled antigens was studied by precipitation of radiolabeled antigen-antibody complexes by anti-rabbit immunoglobulin. Antibodies in sera from rabbits immunized with either BCG (anti-BCG) or melanoma cells (anti-melanoma) bound both the labeled BCG and melanoma antigens. Control antisera, from rabbits immunized with human acute or chronic lymphatic leukemia cells or with normal human spleen cells, did not bind significant amounts of radiolabeled BCG. Antibodies in sera from rabbits immunized with normal spleen cells bound small but significant amounts of radiolabeled melanoma antigens. Binding by anti-BCG and anti-melanoma to the radiolabeled antigens was studied before and after absorption of antisera with cells from human melanoma, leukemia, guinea pig hepatoma, and normal human spleen cells. Inhibition studies using unlabeled BCG extracts also were carried out. The absorption and inhibition studies confirmed that the binding reactions were specific and that antigens from five melanoma patients shared antigenic determinants with BCG.


Cell-mediated immunity to friend virus-induced leukemia. IV. In vitro generation of primary and secondary cell-mediated cytotoxic responses. Primary and secondary cell-mediated cytotoxic responses to FBL-3 cells, a syngeneic Friend virus-induced leukemia in C57BL/6 mice, could be generated by in vitro techniques as tested by the 125IUdR release assay. The specificity of the cytotoxic reactions appeared to be directed against the Friend type-specific antigen and the FMR (Friend, Moloney, Rauscher) antigen which were also the major antigens for transplantation immunity to FBL-3. In comparison to the primary cytotoxic response, the secondary cytotoxic response was accelerated (detected at an earlier time after sensitization), enhanced (gave much higher levels of cytotoxicity), was also longer lasting, and could be induced by a wide dose range of tumor cells. The secondary response could only be induced with lymphocytes obtained from regressors that were resistant to FBL-3 challenge; lymphocytes from mice with progressive tumor growth had no detectable secondary response. It was found that both induction phase and the effector phase of cytotoxic responses were T cell dependent. The characteristics of these reactions were thus very similar to those obtained with in vivo immunization or challenge, providing a good correlation with in vivo tumor immunity.


Studies on autoimmune encephalomyelitis in the guinea pig. II. An in vitro investigation on the nature, properties, and specificity of the serum-demyelinating factor. Complement-dependent demyelinating activity of whole brain homogenate (WBH)-induced experimental allergic encephalomyelitis (EAE) sera was tested on long term tissue cultures of in vitro myelinated fetal guinea pig cerebellum. Complement-fixing (CF) auto-antibodies were shown to be the responsible agents, as demonstrated in experiments where all reagents belonged to the same species: guinea pigs of outbred (Hartley) and even of inbred (S2 or S13) strains. These antibodies were of the IgG2 class as shown by Sephadex G-200 and DEAE cellulose fractionation experiments. The corresponding auto-antigen was present in the homogenate and myelin of the central nervous system (CNS) tissue. It was different from the encephalitogenic basic protein of CNS myelin (BP), as shown in experiments where the demyelinating auto-antibodies were induced, detected, and absorbed by WBH or by CNS myelin but not by BP. They were neither induced by nor cross-reacting with cerebroside and peripheral nervous system (PNS) tissue.


In vivo protection against syngeneic Gross virus-induced lymphoma in rats: comparison with in vitro studies of cell-mediated immunity. Spleen cells at various times after inoculation of W/Fu rats with a syngeneic Gross virus-induced lymphoma, (C58NT)D, were tested for their in vivo activity in adoptive transfer experiments and for their in vitro reactivity in a 4-hr 51Cr release cytotoxicity assay and in a mixed lymphocyte-tumor cell interaction assay. In adoptive transfer, the best protection against tumor growth was observed with immune spleen cells taken at 30 days after tumor cell inoculation (the peak of reactivity in the mixed lymphocyte-tumor cell interaction assay) whereas cells taken at 10 days (the peak reactivity in the 51Cr release cytotoxicity assay) gave only partial protection. The protection detected in the adoptive transfer experiments was specific for (C58NT)D associated antigens, and this correlated well with the specificity observed in the in vitro cell-mediated immunity assays. T cells, but not complement receptor-bearing cells or macrophages, were essential for the protection against tumor growth in vivo, and also for the in vitro reactivity in the 51Cr release cytotoxicity and the mixed lymphocyte-tumor cell interaction assays.


Cell-mediated immune response to herpes simplex virus: type-specific lymphoproliferative responses in lymph nodes draining the site of primary infection. Cell-mediated immune responses to type 1 and type 2 HSV were studied in rabbits using an in vitro lymphocyte transformation assay. The footpad route of inoculation was used to allow us to study the specificity and degree of localization of the responses. Rabbits inoculated in the hind footpads with infectious HSV-I or HSV-II mount type-specific lymphocyte transformation responses that are localized to draining lymphoid organs. Type-specificity requires careful control of all in vitro culture conditions and reflects the extensive cross-reactivity demonstrated by serologic techniques. While lymphocyte transformation responses can be detected with immune SL and PBL, presumably the result of early escape of antigen into the systemic circulation, responses by draining LNL are significantly greater in magnitude. Distant LNL have not been shown to respond. It is postulated that the augmented local immune response to HSV plays a significant role in controlling recurrent HSV infections.


Defining the heterogeneity of anti-tumor antibody responses. Studies were undertaken to analyze the humoral in vivo and in vitro antibody response of BALB/c mice to a syngeneic MSV-induced tumor cell line. With a sensitive radioimmunoassay, sera obtained from individual progressor and regressor mice were shown to vary greatly in total tumor-specific antibody concentration as well as immunoglobulin class distribution of the antibody, but no significant difference existed between the groups of progressor and regressor mice as a whole. In addition, serum antibodies from all animals were shown to have extensive cross-reactivity against a variety of cell lines chosen to share one or more antigens with the cell line used for immunization. Conversely, when in vitro fragment cultures of splenic tissue from progressor and regressor mice were stimulated with tumor-related antigen, differences in responsiveness among normal, progressor, and regressor mice were observed. In addition, antibodies derived from fragment cultures displayed several different cross-reactivity patterns all of which were more restricted in specificity than serum antibodies.


Continuously proliferating T killer cells specific for H-2b targets: selection and characterization. BALB/c (H-2d) thymus-derived lymphocytes sensitized to C57BL/6 (H-2b) alloantigens have been propagated in vitro for over 9 months. These T lymphocytes are specifically cytotoxic to H-2b target cells but are stimulated to proliferate by both H-2b and H-2k spleen cells. This indicates that for these selected cells the antigen requirements for cell proliferation are different from those for cell-mediated cytotoxicity. If not continuously stimulated with allogeneic spleen cells, the cytotoxic cultures fail to divide and rapidly lose their cytotoxic activity. Allogeneic erythrocytes do not stimulate cell proliferation in "quiescent" cell cultures and allogeneic tumor cells do so only in the presence of spleen cells. However, "quiescent" cell cultures display cytotoxicity in the presence of phytohemagglutinin A as do cell cultures which have lost their cytotoxic activity although they proliferate upon allogeneic stimulation. The significance of these findings is discussed.


Specific selection of cytotoxic effector cells: the generation of cytotoxic T cells in rat thoracic duct lymphocyte populations positively or negatively selected for reactivity to specific strong histocompatibility alloantigens. These studies consider the generation of cytotoxic T lymphocytes (CTL) from precursors (CTLP) present in rat thoracic duct lymphocytes after stimulation with strong alloantigens. Also, they explore the relationship between CTLP and "initiator" (I) lymphocytes responsible for specific GVH and MLI reactions. Positively selected TDL populations prepared in bulk MLI cultures show enriched GVH and MLI reactivity for the selecting major histocompatibility complex (MHC) haplotype, but no cytotoxic activity, raising the possibility that I and CTLP may belong to different subpopulations, and the latter failed to differentiate or to survive under these culture conditions. Restimulation of these cells in Marbrook culture vessels with the original priming haplotype under conditions suitable for generating killer cells in vitro resulted in greatly increased specific CTL activity with accelerated kinetics soon after priming and normal kinetics later. These findings indicate that "memory" killer cells can be generated in a previously stimulated lymphocyte population that had no overt cytotoxic activity. Restimulation with third party haplotypes failed to give CTL activity either to specific or to third party targets. Negatively selected TDL populations prepared by "filtration" through x-irradiated F1 rats, depleted of specific GVH and MLI responses, were also depleted of the ability to generate CTL in Marbrook cultures stimulated with the selecting haplotype. Stimulation with third party haplotypes, or with both third party and specific haplotypes together, gave CTL effective only against the third party target.


Specific suppression of the immune response by HGG tolerant spleen cells. I. Parameters affecting the level of suppression. After adoptive transfer, the spleen cells from mice made tolerant to human gamma-globulin (HGG) specifically suppress the immune response of normal spleen cells. However, this suppressive activity in the spleen cells of tolerant mice is only present for a brief period after treatment with tolerogen. Spleen cells from animals injected 10 days earlier with tolerogen reduce the immune response of an equal number of normal spleen cells by 75%. Spleen cells from mice made tolerant 40 days previously are, however, no longer suppressive, even though they remain completely unresponsive. These data suggest that active suppression of antigen-reactive cells is not the mechanism responsible for maintaining tolerance to HGG, but rather is only transiently associated with the tolerant state. Further evidence in favor of the separation of the tolerant state from suppressive activity is that complete suppression of the normal spleen cell response requires either a high ratio of tolerant to immune competent cells or a delay in the antigenic challenge of the reconstituted recipients. By contrast, such manipulations are not required to demonstrate the complete unresponsiveness of the tolerant cells after adoptive transfer.


The inhibition of cutaneous basophil hypersensitivity reactions by a heterologous anti-guinea pig T cell serum. Systemic treatment with a heterologous anti-T cell serum of guinea pigs immunized with EA in IFA markedly suppressed CBH reactivity to specific antigen and T cell mitogens, as judged by gross reactivity, histology, and skin histamine. The antiserum produced a marked drop in circulating lymphocytes, mainly at the expense of T cells, as indicated by the ability of surviving lymphocytes to rosette with rabbit RBC. It was postulated that the suppression of CBH reactivity is due to the depletion of T cells, which would have released a factor chemotactic for basophils. The data therefore provide further evidence that cutaneous reactions rich in basophils are primarily dependent on a population of T cells.


Anti-immunoglobulin stimulation of murine lymphocytes. I. Age dependency of the proliferative response. The in vitro proliferative response of normal mouse spleen cells to anti-immunoglobulin (Anti-Ig) reagents was found to be an age-associated phenomenon. The response usually appears in mice when they reach 5 to 7 months of age and is rarely seen in younger animals. Anti-Ig induced proliferation was observed by using two different antisera--one polyvalent, prepared against mouse antibody-antigen complexes and one prepared against mouse IgM myeloma. Both antisera were shown to be specific for B cells by cytotoxicity and immunofluorescent staining. Glassbead separation of spleen cells showed that the anti-Ig-induced proliferation was a B cell response.


Studies on the antigenic determinants of the Thy-1.2 alloantigen as expressed by the murine lymphoblastoid line S-49.1 TB-2-3. Sialic acid has been shown to be part of the antigenic determinant of the Thy-1.2 alloantigen as expressed by the murine cell line S-49.1 TB-2-3 (S-49). This conclusion is based on the loss of cytotoxic inhibitory activity by the action of neuraminidase on the Thy-1.2 alloantigen. Also, sialic acid has the ability to inhibit the cytotoxic assay of AKR anti-C3H Thy-1.2 serum for S-49 cells. Further evidence for the protein nature of the Thy-1.2 alloantigen is apparent by trypsin digestion. Possibly the Thy-1.2 alloantigen as expressed on S-49 cells is a glycoprotein.


Release of slow reacting substance of anaphylaxis (SRS-A) from human leukocytes by the calcium ionophore A23187. The ability of the calcium ionophore A23187 to release slow reacting substance of anaphylaxis (SRA-A) from human leukocytes was studied. About 25 times more SRS-A activity was released from aliquots of leukocytes by ionophore stimulation than by antigen stimulation, although comparable amounts of histamine were released. Cell separation studies revealed that granulocytes other than basophils were also capable of releasing SRS-A. The contractile activity released after challenge with ionophore appeared physicochemically identical to the SRS-A of rat or human origin released by antigen challenge in terms of its stability to base hydrolysis, inactivation by arylsulfatase, and chromatographic behavior on silicic acid and Sephadex LH-20 columns. We suggest that some mediators of allergic reactions previously associated, in man, only with antigen-IgE antibody interaction on mast cells or basophils may be released by other stimuli and from other cell types.


Suppression of antibody responses in allogeneic mice by products of lymphoid tissue. I. Allogeneic suppressive factor (ASF) from spleens repopulated with thymus cells. A cellfree extract prepared from the spleen cells of C3H mice is capable of suppressing antibody responses to SRBC when extract material is exposed to alloantigens. The observed immunosuppression was attributed to a soluble factor in the extract. This allogeneic suppressive factor (ASF) was detected in extracts prepared from the spleen cells of unirradiated mice as well as those of irradiated mice repopulated with thymocytes, provided that mice were previously immunized with SRBC. Donors of actively suppressive ASF preparations did not need to be previously exposed to alloantigens. Extracts from thymus and marrow cells of unirradiated mice and the spleen cells of irradiated mice repopulated with marrow cells (or no cells) did not contain ASF. C3H thymocytes stimulated with SRBC generated more ASF activity in spleens of C3BF1 hosts than in those of C3H hosts, indicating that alloantigenic stimulation enhances the production or activity of ASF. Once produced, C3H ASF was able to suppress antibody responses in cell transfer experiments only if exposed to C3BF alloantigens of either donor lymphoid cells or irradiated hosts. Once exposed to alloantigens, ASF appears to be capable of suppressing antibody responses of syngeneic C3H or semi-allogeneic C3BF cells. When both donor lymphoid cells and hosts were syngeneic with the donor of the ASF, there was enhancement of antibody formation in cell transfer experiments. C3H ASF did not interfere with education of C3BF thymocytes to SRBC or with the generation of precursors of anti-SRBC antibody-forming cells by C3BF1 marrow cells. ASF may interfere with cellular cooperative events necessary for humoral immune responses or with terminal differentiation of B cells. Production of ASF could partially account for the suppression of antibody responses observed during graft-vs-host reactions.


Suppression of antibody responses in allogeneic mice by products of lymphoid tissue. II. Lack of antigenic specificity and immunogenetic requirements of allogeneic suppressive factor (ASF). Mice were irradiated, infused with thymocytes and immunized with a variety of antigens, i.e., sheep or horse red blood cells (SRBC or HRBC), diphtheria toxoid (DT) or bovine gamma-globulin (BGG). The spleen cells (T.Spleen cells) were harvested 5 days later and cellfree extracts were prepared. The extracts contained an allogeneic suppressive factor (ASF) that was capable of inhibiting IgM antibody responses of allogeneic or semi-allogeneic unirradiated mice. ASF had to be injected within 24 hr of immunization to be effective and a single injection delayed, rather than abolished, the antibody response at the cellular level. However, daily injections of ASF resulted in persistent suppression of antibody response. ASF activity was antigen nonspecific, i.e., the antigen used to stimulate ASF production did not have to be the same as the antigen used to test for ASF activity. C3H T.Spleen extracts were even immunosuppressive when prepared by exposure to C3BF1 alloantigens only; such extracts suppressed antibody responses of C3BF1 and DBA/2 mice. C3H ASF was removed from extracts after incubation with C3BF1 spleen cells but not after incubation with C3H spleen cells. C3BF1 spleen cells which had been preincubated with C3H ASF were unable to generate antibody-forming cells upon transfer to irradiated C3BF1 host mice. This suggests that the ASF molecule may be or include receptors for alloantigens. The immunogenetic requirements for ASF activity were evaluated by injecting extracts from C3H, C57BL, C3BF and BALB/c T.Spleen cells into C3H, CBA, C57BL, BALB/c, DBA/2, A or C3H.A recipient mice. All extracts tested had ASF activity. However, all allogeneic recipients were not suppressed by the extract material. The suppressive activity of ASF seemed to require two (or more) antigenic differences between donors and recipients of extract material, an H-2K or I antigen difference and a second antigen difference, possibility Ig-1. In the limited numbers of strain combinations tested, T.Spleen extracts suppressed IgM antibody response only if exposed to H-2 and Ig-1 antigens, e.g., BALB/c (H-2d, Ig-1a) ASF suppressed A (H-2a, Ig-1e) but not C3H.A (H-2a, Ig-1a) or DBA/2 (H-2d, Ig-1c). Separate ASF molecules may react with separate antigens on the cell surface, i.e., with H-2 and gammaG2a. Alternatively, one ASF molecule may react with two structurally associated antigens. If the latter is correct, it is conceivable that the beta2-microglobulin which is non-covalently linked to the major component of H-2 molecules expresses allotypic antigens coded for by Ig-1 and beta2-microglobulin is one of the antigens recognized by ASF.


Suppression of reaginic antibody formation. III. Relationship between immunogenecity and tolerogenicity of hapten-carrier conjugates. From the study of the effect of epitope density on the immunogenicity of haptenated ovalbumin (DNP-OA) it was concluded that the lightly haptenated conjugate, DNP0-5-OA, induced, on the one hand, only low titers of anti-DNP hemagglutinating antibody and no reaginic antibodies to the hapten and, on the other, high reaginic and high hemagglutinating antibody responses to the carrier. The conjugate with a slightly higher degree of haptenation, i.e., DNP2.3-OA, induced both reaginic and hemagglutinating antibodies to both the hapten and the carrier. By contrast, the heavily haptenated conjugate, DNP20-OA, elicited reaginic and hemagglutinating antibodies only against the hapten but not against the carrier. Specific suppression of anti-hapten reaginic antibody formation had been achieved by treatment of mice with a tolerogen consisting of the hapten (DNP) conjugated covalently to isologous gamma globulins (MgammaG). The epitope density of the DNPx-MgammaG conjugates was shown to play a dominant role in determining whether or not the conjugate was tolerogenic. Thus, lightly haptenated conjugates (DNP0.5-MgammaG, DNP1.3-MgammaG or DNP1.9-MgammaG) were not tolerogenic, moderately haptenated conjugates (DNP4.2-MgammaG, DNP8-MgammaG, and DNP 14-MgammaG) were tolerogenic, and heavily haptenated conjugates (DNP32-MgammaG and DNP53-MgammaG) were immunogenic, being capable of priming the recipients for the DNP hapten. Further evidence for the nonimmunogenicity of DNP 8-MgammaG conjugate was inferred from its rate of clearance in tolerized and normal mice. Thus, the half-life of 125I-labeled DNP8-MgammaG in circulation was not significantly different for normal and tolerized mice; it was 3.7 and 3.5 days, respectively, which is within the range of data reported for clearance of normal MgammaG. These results suggest that DNP8-MgammaG was catabolized at a rate similar to that of nonconjugated, isologous MgammaG. Moreover, there was no significant difference in the localization of DNP8-MgammaG in identical difference in the localization of DNP8-MgammaG in identical organs (spleen, thymus, kidney, and liver) of normal and tolerized mice. All the multivalent DNPx-MgammaG conjugates were shown to be able to elicit passive cutaneous anaphylaxis (PCA) reaction on i.v. challenge of rats which had been pre-sensitized i.d. with anti-DNP reaginic antibodies.


Sequential changes in the concentration of specific serum proteins during typhoid fever infection in man. An automated immunoprecipitin system has been utilized to quantitate the concentration of 10 specific proteins in the plasma of man.  Values obtained by this technique are in agreement with the published concentrations for these specific plasma proteins.  This technique was utilized to determine the sequential change s in 10 individual plasma proteins of volunteers exposed to Salmonella typhi.  In those volunteers who developed typical typhoid fever, plasma concentrations of the acute phase proteins, alpha1-acid glycoprotein, alpha1-antitrypsin, and haptoglobin, as well as C3 complement were significantly increased with the onset of febrile illness.  In contrast, the concentration of plasma albumin and tranferrin were depressed while plasma IgM became elevated during early convalescence from this infection.  No significant changes were observed in the plasma concentrations of alpha2-macroglobulin, IgG, or IgA.  In the exposed volunteers who did not become ill, the only significant change was a brief depression of alpha1-antitrypsin.  During typhoid fever the patterns of change for individual plasma acute-phase globulins were different from those reported for patients with hepatitis, myocaridal infarction, or surgery.


Chemotherapy for head and neck tumours. Observations on a twelve-month study. This paper describes a multiagent protocol of chemotherapy suitable for head and neck cancer which has been used at King's College Hospital since 1974. It studies a twelve month period in which 30 patients received chemotherapy and offers details of eight patients who were free of tumour from two to twelve months after completing the treatment. No statistical evaluation is attempted as the follow-up period is so short and the clinical material so diverse.


Uptake of (125I) iododeoxyuridine in cultured rat and human prostate: effects of insulin and testosterone. DNA synthetic activity was monitored in rat and human prostate using [125I] iododeoxyuridine ([125I]UdR). Fresh prostate tissue from 6-week-old rats showed higher incorporation of [125I]UdR than that from 12- or 26-week-old rats. During culture for up to 6 days in the absence of hormones, the incorporation of [125I]UdR fell to a low level for all three age groups. Stimulatory effects were seen when rat prostates were cultured in the presence of insulin (3 mug/ml) and testosterone (10(-7) mol/l), the incorporation on day 4 of culture being commensurate with that of fresh prostrate of the corresponding age. Thus the magnitude of the response was higher for the 6-week-old prostate than that for the other two age groups. A similar age-related pattern of androgen stimulation was observed in experiments in which immature and adult castrated rats were injected daily with testosterone and the freshly removed prostates were incubated with [125I]UdR. Although insulin, by itself, had a stimulatory effect on [125I]UdR incorporation in cultured prostate, the magnitude of the response did not differ in the 6- and 26-week-old prostate tissue. Maximal stimulation was obtained using 25 mug insulin/ml. Tissue from a benigh prostatic hyperplasia was also responsive to insulin in culture but it differed from rat prostate in that increased proliferative activity occurred even in the absence of hormone stimulation. This spontaneous surge in activity during culture tended to mask the stimulatory effects of insulin and testosterone at concentrations of 3 mug/ml and 10(-7) mol/l respectively.


In vitro tolerance induction of neonatal murine B cells. The susceptibility of neonatal and adult B lymphocytes to tolerance induction was analyzed by a modification of the in vitro splenic focus technique. This technique permits stimulation of individual hapten-specific clonal precursor cells from both neonatal and adult donors. Neonatal or adult BALB/c spleen cells were adoptively transferred into irradiated, syngeneic, adult recipients which had been carrier-primed to hemocyanin (Hy), thus maximizing stimulation to the hapten 2,4-dinitrophenyl coupled by Hy (DNP-Hy). Cultures were initially treated with DNP on several heterologous (non-Hy) carriers and subsequently stimulated with DNP-Hy. Whereas the responsiveness of adult B cells was not diminished by pretreatment with any DNP conjugate, the majority of the neonatal B-cell response was abolished by in vitro culture with all of the DNP-protein conjugates. During the 1st wk of life, the ability to tolerize neonatal splenic B cells progressively decreased. Thus, tolerance in this system is: (a) restricted to B cells early in development; (b) established by both tolerogens and immunogens; (c) achieved at low (10(-9) M determinant) antigen concentrations; and (d) highly specific, discriminating between DNP- and TNP-specific B cells. We conclude that: (a) B lymphocytes, during their development, mature through a stage in which they are extremely susceptible to tolerogenesis; (b) the specific interaction of B-cell antigen receptors with multivalent antigens, while irrelevant to mature B cells, is tolerogenic to neonatal (immature) B cells unless antigen is concomitantly recognized by primed T cells; and (c) differences in the susceptibility of immature and mature B lymphocytes to tolerance induction suggest intrinsic differences between neonatal and adult B cells and may provide a physiologically relevant model for the study of tolerance to self-antigens.


Cell interactions in the suppression of in vitro antibody responses. Normal T and immune B lymphocytes interact in a fashion that leads to suppression of the immune response. Normal spleen cells added to cultures of primed spleen cells specifically suppressed both the IgM and IgG secondary antibody response of the primed cells to less than 30% of the response of the immune cells cultured alone. Cell crowding as a possible in vitro artifact was ruled out. The suppression was specific for the priming antigen, even when the specific and nonspecific antigens were included in the same cultures. Suppression required both normal T and immune B cells to be present in culture. We suggest that the immune population produces a signal that can induce normal T cells to become specific suppressor cells. This form of interaction may represent an important regulatory (homeostatic) mechanism in the immune system.


Isolation and properties of the leukocytosis- and lymphocytosis-promoting factor of Bordetella pertussis. The leukocytosis- and lymphocytosis-promoting factor (LPF) of Bordetella pertussis has been isolated to near homogeneity by physical, chemical, and electron microscopical criteria. LPF contains 14.5% nitrogen and is lipid and carbohydrate free. It is apparently composed of four polypeptide subunits. LPF caused leukocytosis and lymphocytosis in "nude" as well as in normal mice. In addition, purified LPF also induced histamine sensitization and hypoglycemia and refractoriness to the hyperglycemic effect of epinephrine. A monospecific LPF antiserum blocked these reactions as well as leukocytosis and lymphocytosis. LPF is clearly distinct from the hemagglutinating pili of B. pertussis.


Inhibition of the Fc receptor of human lymphoid cells by antisera-recognizing determinants of the hla system. The inhibitory effect of HLA antisera on Fc receptors of human lymphoid cells was investigated. The ability of lymphoid cells to form rosettes (FcRFC) with antibody-coated sheep red blood cells and to function as effector cells (K cells) in antibody-dependent cell-mediated cytotoxicity were used as assay systems. We found that antisera recognizing determinants of the HLA-A, -B, and -C series had no effect on FcRFC, while a specific inhibitory effect was observed of antisera probably reacting with determinants identical to or closely associated with those of the HLA-D series. This inhibitory effect was retained in the F(ab')2 fragments. Specific inhibition of K cells was observed with all HLA antisera, but this effect was lost in the F(ab')2 fragments. We conclude that the Fc receptor of rosette-forming lymphocytes may be closely associated with products of the HLA-D region. This is analogous to the association between the Fc receptor and the Ia antigens on murine rosette-forming B lymphocytes.


An autoradiographic identification of Purkinje axon terminals in the cat. The Purkinje axon boutons terminating in nuclei fastigii and interpositus in the cat have been identified after injection of 3H leucine into the cerebellar cortex overlying the nuclei. The animals survived from 4-48 h after injection of the isotope. Semithin and ultrathin sections were coated and exposed for 3 and 14 weeks, respectively. The electron micrographs showed labelling over myelinated axons down to a diameter of 0.8 mum, and over boutons. Fifty labelled boutons were used for identification of the shape of their synaptic vesicles. The statistical analysis including the test for skewness showed that 39 boutons (78%) fall in one group. Most of the synaptic vesicles in this group are elliptical (ration from 1:1.1 - 1:1.7). Slightly ovoid vesicles (ration up to 1:1.3) are frequent, but flattened vesicles (ration above 1:1.7) are relatively few in this group of boutons. 8% of the boutons have a rather homogeneous vesicle population (prevalence of round vesicles, ratio 1:1). Synaptic specializations of Gray's type II or of an intermediate type were found in the boutons belonging to the first group (78% of the boutons). Specializations of Gray's type I were found in the other bouton groups (8% of the boutons).


Early diagnosis of neonatal bacteremia by buffy-coat examination. This study demonstrates the value of the buffy-coat smear examination in the early diagnosis of neonatal bacteremia. It is inexpensive, easy to perform, and relatively efficient. In most situations large numbers of bacteria could be seen in each positive smear. Unfortunately, the presence of a positive buffy-coat smear was often associated with a poor prognosis; more than one half (four of seven) of infants with positive smears died.


Maintenance of plasma-derived proteins at much lower concentrations in the uterine lumen of the rabbit: a kinetic study of passage. Human serum albumin (HSA) and human gamma globulin (HGG) in serum and uterine fluid of nonpregnant rabbits at various times after an i.v. injection (100 mg/kg) were measured by a radial immunodiffusion test using specific antisera. The HSA concentration in uterine fluid rose to a peak at 12 hr when it was 11% of the serum concentration and then declined, whereas HGG reached a peak at 18 hr (3.2% of serum level) and decreased thereafter. The HSA passed 2 1/2 times faster than HGG, but both proteins equilibrated with uterine fluid in about 12-18 hr. Steady state levels of HSA and HGG indicated that uterine fluid: serum ratios were 1:10 and 1:20, respectively. Similar ratios were found for total protein and rabbit serum albumin (1:10) and rabbit gamma globulin (1:20). Therefore, except when there is a local immune response, the uterine lumen contains only about 5% of the serum antibody concentration. Available data in the mouse, rat and dog also indicate disparity between serum and uterine fluid protein levels.


Familial 'partial 9p' trisomy: six cases and four carriers in three generations. Six cases of translocation trisomy for the distal half of the short arm of a number 9 chromosome and four asymptomatic balanced translocation carriers are presented in a three-generation pedigree. The clinical features are remarkably similar to those recently recognized and increasingly reported in full short arm (9p) trisomy and should be considered a modification of the same syndrome. In addition to non-specific mental retardation and short stature, there is, in common, a characteristic facies, including down-turned corners of the mouth, a slightly bulbous nose, moderately large ears, suggestively wide-set eyes with an antimongoloid slant, dysplasia and hypolasis of the nails, clindactyly of the 5th fingers, and abnormal dermatoglyphs. It appears that the 'trisomy 9p syndrome' in its variant forms, including trisomies for more or less than just the short (p) arm, is one of the most common clinical autosome anomalies in humans, exceeded only by trisomy 21 (Down's syndrome) and possibly trisomies of chromosomes 13 and 18.


Virological studies of baboon (Papio hamadryas) lymphoma: isolation and characterization of foamyviruses. In accordance with a memorandum of understanding in cancer research between scientists in the USSR and the USA, virological studies were performed on two baboons from the Russian colony at the Institute of Experimental Pathology and Therapy at Sukhumi where several cases of leukemia have been observed over the past few years. Foamyvirus isolations were made from lymphoid cells of both of these monkeys, one of which had a confirmed case of lymphoma. The isolates were similar to each other serologically and morphologically, possessed characteristics typical of foamyviruses, and cross-reacted to a low level with antibody to simian foamyvirus type 1.


High-resolution autoradiography as a tool for the localization of nucleic acid synthesis and distribution in the mammalian cell nucleus. Several examples of the application of high resolution autoradiography to the study of nucleic acid distribution in ultrathin sections of fixed and plastic embedded or frozen material are presented. Newly-synthesized DNA, labelled by very short pulses of 3H-thymidine is found to be localized throughout the nucleus. In the blastomeres of early mouse embryos developing after fertilization by 3H-thymidine-labelled spermatozoa, the labelled paternal DNA is distributed non-homogeneously in the nucleus. Finally some results obtained by using a cytochemical enzymatic technique for visualization of DNA directly in ultrathin sections of fixed and embedded cells are demonstrated. Concerning the distribution of RNA, perichromatin localization of rapidly transcribed RNA is described in different cell systems. This label can be attributed to perichromatin fibrils. In cells labelled for longer periods of time, sometimes followed by prolonged postincubations, a residual label is always found over the nucleus. Clusters of interchromatin granules are usually only weakly labelled or unlabelled. These structures probably contain a limited amount of rather slowly labelled RNA. The present results are discussed in the context of some biochemical evidence and of the data described by other investigators.


Biologic effects of auto emissions. I. Exhaust from engine with and without catalytic converter. This paper relates to the efficacy of a catalytic converter in reducing the levels of certain pollutants emitted from an automobile engine and to the reduction and/or elimination of gross biologic damages in animals exposed to emissions from an exhaust system containing such catalysts. Groups of rats were exposed to diluted emissions from an automobile engine with and without catalyst. Concomitantly, a comparative experiment was conducted by exposing a group of rats to carbon monoxide alone (575 mg/m3). The parameters measured included hematocrit, serum LDH, GOT, and lysozyme. An elevation in hematocrit was observed in animals of the experiment run without catalyst and in those exposed to carbon monoxide; the use of the catalyst reduced the carbon monoxide levels in the exposure chambers by more than tenfold and prevented these bioeffects from occurring. Serum LDH activity was elevated in the groups of rats in the experiment conducted without catalyst, but no alternation was observed in the animals from the experiment utilizing the catalyst or in those exposed to carbon monoxide alone. The data obtained in this study showed that acute exposure to noncatalytic emissions caused significant alterations in certain biologic parameters. Conversely, the introduction of an oxidizing catalytic converter into the engine exhaust system reduced or prevented such biologic damage.


Sex chromatin determination in formalin-fixed, paraffin-embedded tissues of rats. Distinctive chromatin bodies appearing in interphase nuclei of mammalian female cells have been found in most mammalian species. The positive correlation between sex chromatin masses and mammalian female tissue presents an almost ideal criterion for sex determination of mammalian tissues. Previous studies indicate that this positive correlation exists in fresh and formalin-fixed, paraffin-embedded normal and tumor tissue of humans, as well as in fresh squash preparations of normal and tumor tissues of rodents. In this experiment we have shown that it is also possible to determine sex in formalin-fixed, paraffin-embedded normal and tumor tissue of the rat.


Teratological evaluation of FD&C red no.2-a collaborative government-industry study. I Introduction, experimental materials, and procedures. Because of recent studies indicating possible embryolethality and teratogenicity of FD&C Red. No 2, and ad hoc committee was convened by the Food and Drug Administration to consider these questions. The committee suggested a collaborative study by three laboratories [Food and Drug Administration (FDA), Industrial Bio-Test Laboratories (IBT), and National Center for Toxicological Research (NCTR)] in which Red No. 2 was given at 200 mg/kg body weight, by gavage during days 0-19, 6-15, or 7-9 of gestation. FD*C Red No. 2 was also given at the same dose level via water bottle. Appropriate controls were utilized. FDA used Osborne-Mendel strain rats, IBT used Charles River strain also showed an increase in the same parameter for the same dose level and in addition showed a significant increase in the percentage of resorptions per litter. It was concluded that because of the inherent variation and the absence of an increase in abnormalities or other indications of embryotoxicity, there is reason to doubt that this effect is either biologically significant or reproducible.


Endogenous ecotropic mouse type C viruses deficient in replication and production of XC plaques. Endogenous ecotropic type C viruses were induced by iodedeoxyuridine from nontransformed and chemically or spontaneously transformed clones of the C3H/10T1/2 cell line. Viruses produced by cells of certain transformed clones were N-tropic and formed large XC plaques. In contrast, viruses produced by nontransformed C3H/10T1/2 cells were not detectable in the XC plaque test. These XC- viruses infected mouse cells with high efficiency, as shown by the induction of murine leukemia virus group-specific antigens in infected cells, but virus production, as determined by DNA polymerase-containing particles, was extremely low. Upon growth in certain mouse cells these replication-deficient, XC(-) viruses converted to type C viruses that were similar in XC assays to N-tropic AKR virus (XC+).


Further characterization of the Friend murine leukemia virus reverse transcriptase-RNase H complex. The purified reverse transcriptase-RNase H complex from Friend murine leukemia virus consists of a single polypeptide of 84,000 molecular weight, which after mild protease treatment in vitro or after intentional degradation during the purification procedure allows the generation of several additional polypeptides. Degradation destroys the RNA-dependent DNA polymerase activity with native RNA templates and reduces RNase H but does not affect response to synthetic template primers such as poly (rA)-Oligo (dT). The properties of the intact murine enzyme consisting of a single polypeptide of 84,000 molecular weight are compared to those of the avian alpha subunit and the avian alpha beta enzyme complex. The intact murine enzyme resembles the avian beta-containing enzyme complex and is different from alpha in the following respects: (i) it binds to native RNA templates; (ii) it transcribes native RNA templates into DNA, a reaction which can be inhibited by actinomycin D; (iii) RNase H activity behaves like a processive exonuclease; and (iv) analysis of the RNase H digestion products reveals oligonucleotides approximately four bases in length.


Inhibition of infectious Rous sarcoma virus production by rifamycin derivative. A new rifamycin derivative, rifazone-82 (R-82), an inhibitor of viral RNA-dependent DNA polymerase, is selectively toxic to transformed chicken cells in culture. R-82 has now been shown to possess antiviral activity as well. The relatively nontoxic properties of R-82 to nontransformed cells have permitted the execution of experiments examining the effect of a rifamycin derivative on virus reproduction. Addition of low concentrations of R-82 (15 mug/ml) to cultures soon after Rous sarcoma virus infection prevents the spread of infection thoroughout the culture. This inhibition is not dependent on concomitant cellular transformation as identical results were obtained with cells infected with a transformation-defective Rous sarcoma virus. Addition of R-82 to cultures in which all the cells are infected does not substantially affect the yield of physical particles as measured by RNA-dependent DNA polymerase activity and by (3H) uridine incorporation into viral RNA. However, the infectivity of the progeny virus, as measured by focus-forming ability, is decrreased 95 to 99% by R-82 treatment.


Unintegrated viral DNA is synthesized in the cytoplasm of avian sarcoma virus-transformed duck cells by viral DNA polymerase. We have examined the location, structure, and mechanism of synthesis of unintegrated viral DNA present in fully transformed cultures of avian sarcoma virus-infected duck cells. De novo synthesis of the unintegrated forms several weeks after the initial infection was documented by labeling unintegrated DNA in both strands with 5-bromodeoxyuridine. The unintegrated DNA is synthesized in, and probably confined to, the cytoplasm, and it consists of duplexes of short "plus" strands (ca. 0.5 X 10(6) to 1.0 X 10(6) daltons) and "minus" strands the length of a subunit of the viral genome (ca. 2.5 X 10(6) to 3.0 X 10(6) daltons). The structure of the duplex and the mode of incorporation of density label support the hypothesis that the unintegrated DNA is synthesized from an RNA templated by virus-coded DNA polymerase.


Low-molecular- weight Rauscher leukemia virus protein with preferential binding for single-stranded RNA and DNA. A sensitive nitrocellulose filter assay that measures the retention of 125I single-stranded calf thymus DNA has been used to detect and purify DNA-binding proteins that retain a biological function from Rauscher murine leukemia virus. By consecutive purification on oligo (dT)- cellulose and DEAE-Bio-Gel columns and centrifugation in 10 to 30% glycerol gradients, RNA-dependent DNA polymerase has been separated from a second virion DNA-binding protein. The binding of this protein to DNA was strongly affected by NaCl concentration but showed little change in activity over a wide range of temperature or pH. After glycerol gradient purification, polyacrylamide gel electrophoresis of this protein showed one major band with a molecular weight of approximately 9,800. This protein binds about as well as to single-stranded Escherichia coli or calf thymus DNA or 70S type C viral RNA. The binding to 125I single-stranded calf thymus DNA is very efficiently inhibited by unlabeled single-stranded DNA from either E. coli or calf thymus and by 70S murine or feline viral RNA. Much larger amounts of double-stranded DNA are required to produce an equivalent percentage of inhibition. This protein, therefore, shows preferential binding to single-stranded DNA or viral RNA.


Intracisternal A particles from FLOPC-1 BALB/c myeloma: presence of high-molecular-weight RNA and RNA-dependent DNA polymerase. Intracisternal A particles from the FLOPC-1 line of BALB/c myeloma have been shown to contain high-molecular-weight RNA (60 to 70S) that is sensitive to RNase, alkali degradation, and heat but resistant to Pronase treatment. The intracisternal A-particle RNA contains tract of poly (A) approximately 180 nucleotides long. As shown in a reconstitution experiment, by antigenic analysis of A-particle preparation and the SC cytopathogenicity assay, the 70S RNA was not due to contamination by type C virus particles. The FLOPC-1 intracisternal A particles also possess an endogenous RNA-dependent DNA polymerase. The enzyme required Mn2+ or Mg2+, dithiothreitol, detergent, and four deoxyribonucleoside triphosphates for maximum activity. Enzymatic activity was maximally stimuated by poly (rC)-oligo (dG)12-18 and less with poly (rG)-oligo (dC)10 or poly (rA)-oligo (dT)12-18 as compare with synthetic DNA/DNA duplex templates such as poly (dA)-oligo (dT)12-18. The enzyme can utilize the A-particle endogenous RNA as template as shown by analysis of the early and late DNA products of the endogenous reaction by CsSO4 isopycnic gradient centrifuation and hybridization of purified 70S or 35S A-particle RNA with the purified complementary DNA product. Approximately 50% of the A-particle complementary DNA also hybridized with oncornavirus RNA.


Spontaneous release of endogenous ecotropic type C virus from rat embryo cultures. Type C viruses were isolated from embryo cultures of two different rat strains, Sprague-Dawley and Fischer. Both viruses (termed rat leukemia virus, RaLV) were released spontaneously from rat embryo cells, have a density of 1.14 to 1.15 g/cm(3) based on equilibrium sedimentation in sucrose gradients, contain 60-70S RNA, RNA-directed DNA polymerase, and rat type C virus-specific 30,000 molecular-weight-protein determinants. Molecular hybridization studies using the Sprague-Dawley RaLV 60-70S RNA show that the virus-specific nucleotide sequences are present in the DNA of rat embryos. Both Sprague-Dawley and Fischer RaLV can rescue the murine sarcoma virus genome from Kirsten murine sarcoma virus-transformed nonproducer cells and are neutralized by antisera to the RPL strain of RaLV. In contrast to previous RaLV's, these viruses propagate in their own cells of origin as well as in cells of heterologous rat strains.


Intracellular restriction on the growth of induced subgroup E avian type C viruses in chicken cells. Subgroup E avian type C viruses produced by bromodeoxyuridine-treated 100 X 7, line 7, or line C chicken cells were restricted in their intracellular growth on K28 chicken cells but not on line 15 chicken cells. Cells from embryos of line 15 chickens bred with K28 chickens did not restrict the growth of the subgroup E induced leukosis viruses (ILVs). This result indicates that the phenotype for the intracellular restriction of the growth of subgroup E ILVs found in K28 cells is recessive. Long-term growth of the subgroup E ILVs in K28 cells resulted in the appearance of subgroup E virus that grew well on K28 cells. No change in growth characteristics was observed for subgroup E ILVs grown in line 15 cells indicating that appearance of nonrestricted virus occurred only during growth of the subgrouo E ILVs on a restrictive host. RAV-0, a subgroup E virus closely related to the ilvs, had the same growth characteristics as the subgroup E ILVs. RAV-60, a subgroup E virus formed by recombination of exogenous avian leukosis virus with endogenous subgroup E virus coat information, grew well on both line 15 and K28 cells.


Characterization of the oncornavirus particles in the plasma of guinea pigs with L2C leukemia. The inoculation of L2C guinea pig leukemia cells into strain 2 guinea pigs results in the death of the animals within 12 to 15 days. Death is preceded by the simultaneous appearance in the plasma of (i) elevated leukocyte levels, (ii) extracellular virus particles, and (iii) a particle-associated RNA-directed DNA polymerase. This enzyme activity has a cation preference identical to that of the type B bromodeoxyuridine-induced guinea pig virus, i.e., an Mg2+ optimum at 20 mM and no activity using Mn2+. Competitive molecular hybridization studies also revealed that the plasma of leukemic guinea pigs contained approximately 2 X 10(9) genome equivalents per ml of an RNA that is homologous to the RNA of the bromodeoxyuridine-induced guinea pig virus. Morphological observations indicate that most, but not all, of the extracellular particles observed in leukemia plasma are derived from the intracisternal particles seen in the L2C tumor cells. The possibilities that either two viral populations are present or that the in vivo morphogenesis of the type B bromodexoyuridine-inducible guinea pig virus is markedly different from its in vitro morphogenesis are discussed.


Mechanism of release of active alpha subunit from dimeric alpha beta avian myeloblastosis virus DNA polymerase. Storage of the dimeric (alphabeta) form of avian myeloblastosis virus (AMV) DNA polymerase in glycerol resulted in the release of the smaller alpha subunit, as detected by glycerol gradient sedimentation. Analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis of enzyme stored in glycerol showed the concomitant appearance of several polypeptides and a lowering in the level of both beta and alpha components. This reduction appears to be the result of cleavages introduced by traces of hydrolytic activity present in glycerol samples. An enhancement of alpha subunit released, as detected by activity profile, was also achieved upon direct but limited exposure of purified avian myeloblastosis virus DNA polymerase to carboxymethyl-cellulose-bound trypsin matrix. Electrophoretic analysis of digested enzyme revealed a progressive fragmentation, with simultaneous increase in the alpha subunit and decrease in the beta subunit.


Radiommunoassays for the 70,000-molecular-weight glycoproteins of endogenous mouse type C viruses: viral antigen expression in normal mouse tissues and sera. Genetic information coding for type C RNA viruses is transmitted within the DNA of mouse cells. At least three endogenous viruses have so far been immunologically distinguished by radioimmunoassays for their 12,000-molecular-weight polypeptides (p12). In the present study, the 70,000-molecular-weight glycoproteins (gp70) of three prototype viruses were purified, and competition radioimmunoassays were developed for each. By use of these immunoassays, the antigenic determinants of gp70's of different classes of endogenous virus, isolated from the same and from a variety of other mouse strains, were readily discriminated. In contrast, viruses of the same class were indistinguishable. These findings further document the existence of three distinct endogenous viruses of mouse cell. The levels of type C viral gp70 were quantitated in tissues and sera of several inbred strains. The pattern of immunological reactivity of the gp70 detected in serum was indistinguishable from that of the viral gp70 partially purified from tissues of the same strain. Moreover, in each case it was indistinguishable from that of a specific class of endogenous virus. In virus-negative tissues of BALB/c and NIH Swiss mice, the viral gp70 detected was shown to be representative of a class III endogenous virus whose p12 polypeptide was also expressed by the same cells.


Detrusor hyperreflexia in benign infravesical obstruction. a cystometic study. Cystometry was performed on 94 consecutive patients with benign prostatic hypertrophy or bladder neck obstruction, 45 per cent of whom had detrusor hyperreflexia. Of those cases with detrusor hyperreflexia 21 patients were re-examined 6 months postoperatively and 62 per cent of them had regained normal bladder function. The underlying pathophysiologyand the clinical aspects are discussed.


Immunological: reactions involving leukocytes: III. Agranulocytosis induced by antithyroid drugs. A method has previously been described which detected xenogeneic and allogeneic antibodies to human granulocytes by their inhibition of the normal phagocytosis-associated hexose monophosphate shunt (HMS) activity. This method was used to study three patients with acute agranulocytosis secondary to antithyroid drug administration. Two patients with methimazole and one patient with propylthiouracil induced agranulocytosis were studied. Serum samples from each of these three patients taken during the acute phase of agranulocytosis had inhibitory effects on phagocytosis-associated HMS activity in leukocytes from both normal donors and the patients after their full recovery from agranulocytosis. IgM but not IgG prepared from acute sera in two patients was also inhibitory. Disruption of IgM disulfide bonds by dithiothreitol destroyed its inhibitory activity. The possibility of drug-dependent immune destruction of leukocytes in these patients is discussed.


[Recent details of the frequency and immunologic reactivity of sympathetic ophthalmia (author's transl)]. When giving details of the frequency of Sympathetic Ophthalmia a clear distinction must be made between this disorder and the Phacogenic Ophthalmia also affecting the second eye. The latter is found with lens damage, whereas genuine Sympathetic Ophthalmia in relation to enucleation, eye damage and eye opening surgery at Heidelberg University between 1937 and 1973 is less than 1%, a figure markedly below figures quoted in the earlier literature. In two out of three patients with recent genuine Sympathetic Ophthalmia we found an augmented stimulation index in the LTT with retina antigen, not choroid antigen, as compared to control groups. This can be regarded as due to an individually augmented immunologic reactivity towards eye specific tissue antigens.


[Alpha-fetoprotein and cystic fibrosis (author's transl)]. Recently significantly raised serum levels of alpha-fetoprotein (AFP) were demonstrated in patients with cystic fibrosis (CF) and in their parents and some siblings; thereupon the determination of AFP was recommended for detecting heterozygote carriers of the CF-gene. These results could not be confirmed by our own investigations also using a radioimmunassay.


Prolongation of postoperative disease-free interval and survival in human colorectal cancer by B.C.G. or B.C.G. plus 5-fluorouracil. 83 patients with colorectal carcinoma of the Dukes' C class were randomised to receive postoperative adjuvant therapy with B.C.G. alone or in combination with oral doses of 5-fluorouracil (5-F.U.), and have been followed for up to thirty months. Results were compared with carefully selected historical controls who were treated by surgery alone. A statistically significant prolongation of both disease-free interval and overall survival was observed in 50 patients receiving the combination of B.C.G. and 5-F.U. (P=0.03, P=0.01 respectively) as well as in 33 patients receiving B.C.G. alone (P=0.03, P=0.05 respectively). The efficacy of B.C.G.+5-F.U. was independent of the number of tumour-involved lymph-nodes in the surgical specimen. In contrast, B.C.G. given alone appears to be highly effective among 10 patients with 6 or more positive lymph-nodes (P less than 0.04) and ineffective (as yet) among 23 patients with 5 or less positive lymph-nodes. These results suggest that adjuvant immunotherapy, with or without chemotherapy, can improve the prognosis of surgically treated patients with colorectal carcinoma of the Dukes' C class.


Cold and warm antibodies and graft survival in kidney allograft recipients. Serial serum samples from 35 renal allograft recipients were screened for cold and warm cytotoxic antibodies using a two-stage microlymphocytotoxicity test with three incubation temperatures. Both cold and warm antibodies were reactive at 22 degrees C. The critical temperatures which distinguished the two classes of antibodies were 15 degrees C and 37 degrees C. Cold antibodies were detected in sera from 15 patients, 10 patients had warm antibodies, and the remaining 10 had a mixture of cold and warm antibodies. All the patients were cytotoxin-positive after transplantation, although not all had preformed lymphocytotoxins. At one year 14 of the patients with cold autoantibodies had functioning grafts, but only 3 of the 10 patients with warm antibodies had kidneys functioning. This difference was statistically highly significant. There was no difference in graft survival among patients with warm antibodies and those with a mixture of cold and warm antibodies.


Acute platelet changes after large meals of saturated and unsaturated fats. Large amounts of saturated fats (S.F.) or unsaturated fats (U.S.F.) were given to healthy volunteers at a single meal. The heparin thrombin clotting-time, which may measure platelet factor 4 released from platelets into the plasma, was shortened after S.F. and prolonged after U.S.F. The antithrombin clotting activity decreased after S.F. and increased after U.S.F. The platelet-count decreased and the platelet volume increased after both S.F. and U.S.F.


Reimplantation of a human embryo with subsequent tubal pregnancy. A human embryo in transition between a morula and blastocyst after culture in vitro was reintroduced into the mother's uterus via the cervix. The resulting pregnancy was closely monitored and was found to be located in the oviduct. The ectopic embryo was removed at 13 weeks gestation.


Perception of asthma.

Perception of asthma. Subjective assessment and objective measurements of airways obstruction were compared in 82 patients during methacholine-induced asthma. 15% of the patients were unable to sense the presence of marked airways obstruction (forced expired volume in 1 s less than 50% of the predicted normal value). These subjects could not be characterised as a distinct group on the basis of their sex, age, or duration of their asthma. This reinforces the need for objective measurement of lung function in the management of asthma.


A fatal motor-car accident and cannabis use. Investigation by radioimmunoassay. Impairment of driving skills by drugs is an important cause of traffic accidents. Alcohol is the most important, though far from the only, drug involved; and of 684 fatal accidents investigated by Woodhouse, 321 (47%) of the drivers had blood-alcohol levels greater than 100 mg/100 ml at the time of death. In 16 it exceeded 400 mg/100 ml. Unlike alcohol, cannabis has received little attention as a possible cause of traffic accidents, largely owing to the difficulty of proving cannabis use objectively. The recent development of a reliable and relatively simple method for detecting and measuring cannabis products in blood and urine may help to overcome this difficulty. As Milner has pointed out, the full effect of alcohol on driving competence was not appreciated until objective methods of measuring blood-alcohol levels became generally available. The same may be true of cannabis.


Cancer of the uterine corpus after hormonal treatment for breast cancer. Among 45853 women in whom breast cancer was diagnosed after age forty-nine, from the series of the End Results Program of the National Cancer Institute, cancer of the uterine corpus subsequently developed in 203. The risk was greater among those women receiving hormones than in other treatment groups, and tended to rise with increasing interval from first treatment. One method of estimating an expected value indicated that the excess risk of corpus cancer in breast-cancer patients was restricted to those treated with hormones. Given the time period under study, it may be assumed that the hormones were primarily non-steroidal oestrogens.


Colonic involvement in salmonellosis.

Colonic involvement in salmonellosis. Although small intestine is generally regarded as the principal site involved in human salmonella infection of food-poisoning type, the pathophysiology of salmonella diarrhoea remains obscure. A sigmoidoscopic and biopsy study of 23 hospital inpatients with acute salmonella infection revealed presence of active colitis in 21. It is suggested that colonic involvement is common in human salmonellosis and probably plays an important role in the causation of diarrhoea.


Release of motilin after duodenal acidification. The physiological stimulus leading to motilin release in man has not yet been determined. Six healthy volunteers had 3-minute intraduodenal infusions of acid, alkali, or saline solutions. After infusing acid, plasma-motilin rose by 90% at 4 min and was still significantly raised at 45 min, whereas there was an insignificant fall in plasma-motilin after alkali and no change after saline solution. Release of motilin after duodenal acidification may be an important factor in inhibiting gastric emptying.


Dietary iodine and risk of breast, endometrial, and ovarian cancer. Geographic differences in the rates of breast, endometrial, and ovarian cancer appear to be inversely correlated with dietary iodine intake. Endocrinological considerations suggest that a low dietary iodine intake may produce a state of increased effective gonadotrophin stimulation, which in turn may produce a hyperoestrogenic state characterised by relatively high production of oestrone and oestradiol and a relatively low oestriol to oestrone plus oestradiol ratio. This altered endocrine state may increase the risk of breast, endometrial, and ovarian cancer. Increasing dietary iodine intake may reduce the risk of these cancers.


Evaluation of quality of life in patients receiving treatment for advanced breast cancer. Linear analogue self-assessment was used to measure the subjective effects of treatment in women with advanced breast cancer. The results show that this technique may be used to monitor the subjective benefit of treatment and to compare the subjective toxicities of different therapeutic regimens. The results also suggest that the subjective toxicity of cytotoxic therapy is not related to the patient's age and diminishes with successive courses of drugs.


Endoscopy and routine and double-contrast barium meal in diagnosis of gastric and duodenal disorders. A double-contrast barium meal has been carried out on 37 patients in whom there had been disagreement in diagnosis between the routine barium meal and subsequent oesophago-gastro-duodenoscopy. There was an 81% agreement between the endoscopic diagnosis and the diagnosis obtained with double-contrast barium meal. Confirmation of the endoscopic and double-contrast radiological diagnosis was obtained all 11 patients who had surgical treatment. These findings suggest that a double-contrast barium meal should be routinely used in the diagnosis of dyspepsia.


Florid and subclinical rickets among immigrant children in Glasgow. 200 Asian 300 African, Chinese, or Scottish children were examined for clinical, biochemical, and radiological evidence of vitamin-D deficiency. Among the Asians there were 10 with florid rickets and 15 with subclinical rickets. Among the African, Chinese, and Scottish children there were no cases of florid rickets and only 7 cases of subclinical rickets (3 African, 3 Chinese, and 1 Scottish). Loss of metaphyseal definition is considered to be the radiological pattern of minimal active rickets and metaphyseal bands to represent the healing stage. Serum 25-hydroxycholecalciferol concentrations were low in clinical and subclinical minimal active rickets although there was some overlap with the wide range found in the "normal" group. Elevated serum-alkaline-phosphatase levels alone should not be regarded as indicating vitamin-D deficiency. The continuing prevalence of rickets in Asian children and in particular among schoolchildren warrants immediate action, which is long overdue.


Prevention of vitamin-D deficiency in Asians. Asian families living in Glasgow were studied between December, 1973, and June, 1974. One group of families served as a control; their mean serum-25-hydroxy-vitamin-D (25-hydroxy-cholecalciferol and 25-hydroxyergocalciferol) remained low, being 5-1 +/- 0-8 (S.E.M.) ng/ml at the end of the study. A second group was given 3000 units of vitamin D2 in a capsule weekly; this raised the mean concentration of 25-hydroxy-vitamin-D to 18-1 +/- 2-9 ng/ml. The variable effectiveness of this supplement was attributable to some subjects not taking the capsules regularly. The third group of families was provided with chupatty flour fortified with vitamin D(6000 units per kg). This increased mean serum-25-hydroxy-vitamin-D very uniformly to a mean value of 19-5 +/- 1-2 ng/ml. It is concluded that vitamin-D deficiency in Asian immigrants could be substantially reduced by fortification of chupatty flour with vitamin D.


Double-blind clinical trial of anaesthetic premedication for use in major day surgery. Three anaesthetic premedication regimens have been compared by double-blind controlled trial in 158 patients undergoing day-case surgery for varicose veins or hernia. Atropine plus droperidol was superior to atropine plus diazepam or atropine alone in lessening nausea and vomiting and in reducing the need for postoperative analgesia.


Efficacy of low-dose heparin in prevention of extensive deep-vein thrombosis in patients undergoing total-hip replacement. In a prospective trial, the prophylactic efficacy of low-dose heparin was investigated in 84 consecutive patients undergoing total-hip replacement. 52 of these were randomly allocated to receive heparin or a combination of heparin and dihydroergotamine (D.H.E.); the remaining 32 patients, who were operated on by one orthopaedic team and did not receive prophylaxis, acted as a control group. The frequency of deep-vein thrombosis (D.V.T.), determined by the 125I-fibrinogen test and venography, was 69% in the control group, 32% in the patients receiving heparin alone, and 16% in those receiving heparin and D.H.E. The difference between the control group and each group receiving prophylaxis was statistically significant. The frequency of femoral-vein thrombi, demonstrated by venography, was analysed separately because of its frequent association with major pulmonary embolism. Femoral-vein thrombi developed in 17 (53%) in the control group, while they developed in only 4 patients (8%) in the prophylaxis group (P less than 0.01). 1 patient in the control group died due to massive pulmonary embolism; in another patient, again in the control group, clinical features suggestive of major non-fatal pulmonary embolism developed. This complication was not observed in patients receiving prophylaxis. There was no difference in the amount of operative or postoperative blood-loss in the three groups. In 3 patients in the entire series wound haematomata developed; all 3 were controls and haematomata developed after the administration of streptokinase or large doses of heparin for the treatment of extensive femoral-vein thrombi. By use of a sensitive assay, heparin could be detected in the plasma in 67% of samples in patients in whom D.V.T. did not develop. In contrast, in those patients in whom D.V.T. developed, heparin could be detected in only 21% of blood-samples (P less than 0.01). It is concluded that in patients undergoing total-hip replacement, low-dose heparin prophylaxis is effective in reducing the frequency of life-threatening extensive thrombi in the femoral vein. Prophylaxis using a combination of heparin and D.H.E. requires further evaluation.


Bromocriptine treatment of secondary amenorrhoea. Bromocriptine 2-5 mg twice daily is effective in the treatment of both normoprolactinaemic and hyperprolactinaemic secondary amenorrhoea. This was demonstrated by the restoration of menstrual cycle and/or ovulation in 9 of 18 normoprolactinaemic and in 8 of 14 hyperprolactinaemic patients taking bromocriptine. Serum-prolactin level decreased in both groups of patients, and usually menstruation was recovered within 8 weeks'treatment. Galactorrhoea disappeared in 7 of 9 hyperprolactinaemic patients, and 2 became pregnant. After treatment had ceased spontaneous menstrual activity continued in 4 patients. 16 patients had side-effects the commonest being nausea and vertigo. These usually disappeared with the dosage was reduced, but 5 patients refused to continue. These results point to a new approach in the treatment of secondary amenorrhoea, even in those patients whose clinical findings give no indication of prolactin suppression.


Copper incorporation studies on cultured cells for prenatal diagnosis of Menkes' disease. The incorporation of 64Cu into cultured fibroblasts from 7 boys with Menkes' steely hair disease and from 9 controls was examined. The median 20 h incorporation was 74-4 ng 64Cu/mg protein for the patients and 26-1 ng 64Cu/mg protein for the controls (P less than 0.01). Thus, the defect in copper metabolism in the disease was expressed in cultured cells. The method was applied to amniotic fluid cell-cultures of 2 pregnancies at risk and to amniotic fluid cell-cultures with a male karyotype of 8 normal pregnancies. The median 20h 64Cu incorporation into the normal cell-cultures was 19.2 ng 64Cu/mg protein, into the cell-culture from one pregnancy at risk was 48-6 ng 64Cu/mg protein, and from the other, 12-5 ng 64Cu/mg protein. The first case was aborted and the copper distribution in the fetus was found to be abnormal--consistent with Menkes' disease. The second case at risk was born in May, 1975, and no signs of Menkes' disease have developed. These cases demonstrate that prenatal diagnosis of Menkes' steely hair disease might be feasible.


Evidence for cell-mediated immunity to renal antigens in minimal-change nephrotic syndrome. A direct lymphocytotoxicity test was used to investigate 12 patients with minimal-change nephrotic syndrome and 12 patients with proliferative glomerulonephritis for cell-mediated immunity against an epithelial cell culture derived from human kidney. Lymphocytes from patients with minimal-change nephritis but not from those with proliferative disease had significantly greater lymphocytotoxicity against the cultures than did lymphocytes from normal controls.


Inhibition of lymphocyte blastogenesis by plasma of patients with minimal-change nephrotic syndrome. Plasma from 7 patients with minimal-change nephrotic syndrome (M.C.N.S.) (lipoid nephrosis) confirmed by renal biopsy inhibited the blastogenic response of lymphocytes to the mitogen phytohaemagglutinin (P.H.A.) and to allogeneic lymphocytes in mixed lymphocyte culture. Plasma from 14 patients with nephrotic syndrome due to other glomerular diseases did not have this effect. 5 of the 7 patients with M.C.N.S. achieved remission; 4 with prednisone and 1 spontaneously. While in remission, plasma from these 5 patients no longer inhibited the response of lymphocytes to P.H.A. These findings suggest a pathogenic relation between M.C.N.S. and cell-mediated immunity.


Metabolic inhibition of polymorphonuclear leucocytes in fulminant hepatic failure. A high incidence of bacterial infections in patients with fulminant hepatic failure (F.H.F.) has led to an investigation of polymorpho-nuclear-leucocyte (P.M.N.) function. No intrinsic leucocyte abnormality was demonstrable but a factor present in F.H.F. serum was shown to inhibit the metabolic activity of the leucocyte hexose-monophosphate shunt. This effect was due neither to low serum-complement nor to associated renal failure. The inhibitory factor, however, was removed either by pre-incubation with activated charcoal or by in-vitro dialysis, raising the possiblity that charcoal haemoperfusion or other forms of artificial liver support may improve P.M.N. function in this condition.


Mechanism for increased production of prostaglandins in labour. It is proposed that uterine hypoxia, beginning initially with the uterine contractions in late pregnancy, leads to the increased formation of free fatty acids. The liberated fatty acids are then used to form prostaglandins and they can also act on mitochondria to release calcium. The released calcium can directly activate contractile proteins and/or can further stimulate phospholipase A (active at neutral pH) and thereby reinforce the progressive production of prostaglandins.


Death in acute upper gastrointestinal bleeding. Can endoscopy reduce mortality? In a retrospective analysis of 64 deaths from acute upper gastrointestinal bleeding in Edinburgh Royal Infirmary, 46 of the patients (72%) were found to have been over 60 years of age. Recurrent or continuous bledding occurred in 24 cases; after exclusion of deaths from sudden exsanguination (9) and from bleeding varices with subsequent hepatic failure (11), only 4 patients died from recurrent bleeding. Other causes of death were postoperative complications (20) and other severe concomitant disease (20). These results suggest that improved diagnostic techniques such as endoscopy may not significantly reduce the mortality from acute upper gastrointestinal bleeding.


Controlled oxygen therapy at 60% concentration. Why and how. In a number of circumstances pulmonary function is so disturbed that inspired oxygen concentrations of about 60% are needed to combat arterial hypoxaemia. Rational therapy requires that the inspired concentration be controlled and not vary with the patient's breathing. An exploration of the interaction between gas flow, ventilation, breathing pattern, and mask volume shows that a mask of 300 ml volume supplied with a flow of 30 1/min (15 1 O2, 15 1 air) can meet the need.


Metabolic fate of administered 1,25-dihydroxycholecalciferol in controls and in patients with hypoparathyroidism. In four healthy controls and three patients with hypoparathyroidism serum-1,25-dihydroxycholecalciferol (1,25-D.H.C.C.) concentrations, after oral or intravenous administration, declined biphasically with a rapid-phase half-time of about 14 hours. Repeated oral doses of 1 mug 1,25-D.H.C.C. (2-4 nmol) produced serum concentrations well below the assayed normal range but were nevertheless effective in raising serum-calcium. It is suggested that orally administered 1,25-D.H.C.C. acts directly on the intestinal mucosal-cell nucleus to promote calcium absorption. 1,25-D.H.C.C. is more rapidly eliminated from the body than vitamin D, and it is predicted that any hypercalcaemia caused by 1,25-D.H.C.C. therapy should be of relatively short duration.


Relation of thyroid hormones to body-weight. Serum-triiodothyronine is positively and significantly correlated with body-weight. It is increased by overeating and reduced after weight-loss. No correlation has been found between serum-thyroxine and body-weight or changes in body-weight.


Evaluation of three thyroid-function screening tests for detecting neonatal hypothyroidism. Three thyroid-function detection methods were evaluated concomitantly for possible application as routine screening tests for the early diagnosis of neonatal hypothyroidism. Dried capillary-blood thyroxine (T4) was measured on eluted filter-paper discs for 9734 3-5-day old neonates. Serum T4 and thyroid-stimulating hormone (T.S.H.) assays from cord blood were studied as alternative screening tests on 4911 and 3733 infants, respectively. To avoid false-negative results, neonatal blood-T4 and cord serum-T4 screening methods were followed up for the lower 10th and 6th percentile, respectively. This resulted in a false-positive recall incidence greater than 92% owing to various additional factors which also influence T4 levels: thyroxine-binding-globulin deficiency, prematurity, and maternal drug ingestion. In marked contrast, cord T.S.H. as an initial screening test had a higher specificity and sensitivity for the diagnosis of primary hypothyroidism with the two affected cases, having values greater than 70 muU/ml (with only 0-24% of the screened population having values greater than 50 muU/ml). In order to avoid the impracticably high recallrate and false-positive incidence resulting from an initial T4 screening test, and to reduce the estimated follow-up recall to less than 0-2% of the screened infant population, it is recommended that infants with low T4 be selected for a supplementary T.S.H. screening test.


Treatment of rosacea by metronidazole.

Treatment of rosacea by metronidazole. A double-blind trial in twenty-nine patients with rosacea showed that, after 6 weeks' treatment, metronidazole was therapeutically superior to a placebo (P less than 0-02). It was particularly effective against papules and pustules. The mode of action of metronidazole and other antibiotics in rosacea is not known.


A five-year follow-up of Lord's dilatation for haemorrhoids. 100 consecutive patients treated by Lord's dilatation for haemorrhoids were followed up for a mean of 5-1 years. 2 patients could not be traced, and 1 had died. 75 patients were symptom-free or greatly improved. For 22 patients the treatment was unsatisfactory and 19 have since undergone haemorrhoidectomy. The outcomes at six months and five years were the same in 87%. The results of this follow-up study indicate that treatment of haemorrhoids by Lord's dilatation produces satisfactory long-lasting results in most patients.


Role of gluten, prednisone, and azathioprine in non-responsive coeliac disease. A case of severe "non-responsive" coeliac disease has been shown to be sensitive to gluten whilst under prednisone therapy, but not without prednisone. This adds an additional dimension to the criteria for diagnosis of coeliac disease. Cases of non-responsive coeliac disease may require both prednisone and gluten exclusion to induce a remission. In this patient, azathioprine induced and sustained a remission when unacceptably high doses of prednisone had failed, and may prove to be a valuable immunosuppressive in non-responsive coeliac disease.


Rheumatoid arthritis and alpha-1-antitrypsin. The genetic types of alpha1-antitrypsin (Pi types) were studied in 55 adults with classical or definite rheumatoid arthritis and 56 children with juvenile rheumatoid arthritis. A comparison was made with control groups of 211 healthy adults and 512 schoolchildren. Pi types MZ and SZ were significantly more prevalent in adults with rheumatoid arthritis than in the control adults. There was no difference between the patients with juvenile rheumatoid arthritis and the control children. Reduced concentrations of alpha1-antitrypsin, as associated with heterozygotes for the Z allele, may be a contributory factor to the development of rheumatoid arthritis or to the tissue destruction of rheumatoid arthritis.


Pathogenesis of essential hypertension.

Pathogenesis of essential hypertension. It is proposed that essential hypertension is an exaggeration of the tendency for blood-pressure to rise with age, that it results from a repeated sequence in which a small rise in pressure, possibly resulting from an autonomic nervous overactivity, produces changes in the kidney which maintain the rise of pressure and become the basis for a further rise.


Mechanism of renal hypertension.

Mechanism of renal hypertension. Renal hypertension of the two-kidney type is divided into three stages. In the first, hypertension results from the vasoconstrictor effect of angiotensin II. This persists to some extent in the second phase but there is in addition a slow-developing pressor effect, also resulting from angiotensin II and probably attributable to sodium. In the first two phases removal of the abnormal kidney corrects the hypertension. This fails in the third phase because changes in the opposite kidney maintain hypertension. Renin and angiotensin are probably not involved at this stage.


A long-term controlled trial of screening for hypertension in general practice. A controlled trial was undertaken to evaluate the practical consequences of screening in general practice. Of 2420 people aged 40-64 years examined in 1967-68, 191 (7.9%) had previously been told, at some time, that they had raised blood-pressure, other than during pregnancy. However, only 77 (3-2%) had any record of current antihypertensive treatment. Screening resulted in a further 50 persons (2-1%) being newly diagnosed as hypertensive in 1967-68 and a further 9 in 1969-70. Antihypertensive treatment was given to 21 of these immediately following diagnosis, while the treatment was adjusted in 23 of those already known to be hypertensive. In 1972-73 the blood-pressures of the screening population were directly compared with the control group. No significant differences in the distributions of their blood-pressure levels were observed. Over 95% of the new hypertensives discovered by the screening process in the control group in 1972-73 had visited their general practitioners for some reason during the previous five years. This suggests that "case-finding" by general practitioners would be more cost-effective than setting up separate blood-pressure screening clinics. However, the results of this study indicate that we need to know more about how raised blood-pressure can be successfully controlled over a long time, before any mass screening programmes can be actively encouraged.


Parity of women contracting rubella in pregnancy. Implications with respect to rubella vaccination. Data from the National Congenital Rubella Surveillance Programme showed that 44% of children with congenital rubella reported to the programme were born to primiparae. This high proportion is thought to be due to the fact that there was a two-fold increase in the rate of abortion for rubella in pregnancy for women with two or more children. This higher incidence of congenital rubella in firstborns emphasises the need for rubella vaccination prior to a woman's first pregnancy.


Anaemia and immune response.

Anaemia and immune response. Nutritional anaemia is a major public-health problem in many parts of the world, and iron deficiency appears to be the most important cause. The immune response is believed to be impaired in anaemia. The results of the present study, carried out in young children, indicate that both the cell-mediated immune response and the bactericidal activity of leucocytes are impaired when levels of haemoglobin fall to 10 g/dl or less.


WRL 105 strain (H3N2) live attenuated influenza vaccine: acceptability, reactivity, and antibody response in normal, bronchitic, and geriatric volunteers. The acceptability, reactivity, and antibody responses of recombinant WRL 105 strain, live, attenuated influenza virus vaccine administered intranasally were studied in seventeen normal adults, and in seventeen bronchitic and twenty-one geriatric volunteers. The effect on peak flow and 1-second forced expiratory volume (F.E.V.1) on the 3rd, 5th, and 7th days after vaccination was measured in the bronchitic and normal groups. Seroconversion occurred in 80% to tht homologous virus, in 40.6% to A/Victoria/3/75, and in 26.5% to A/England/864/75 in subjects with pre-vaccination haemagglutination inhibition titres of less than 1/40. A fourfold or greater increase in homologous anti-neuraminidase antibody was found in 48% of twenty-seven infected subjects when measured by a new elution inhibition technique. The frequency and nature of symptoms were similar in both infected and non-infected groups. No significant changes in F.E.V.1 occurred, but on days 5 and 7 there was a decrease in peak flow measurements in both infected and non-infected groups when assessed as the percentage change of the pre-vaccination value.


Evidence of an agent transmissible from ulcerative colitis tissue. Five New Zealand White rabbits were injected intracolonically with homogenates (100 mum) of ulcerative-colitis tissue. Histological changes closely similar to those seen in the human donors were present in the mucosa and submucosa of the large intestine of four of these rabbits 3-12 months later. Similar changes were seen in the large intestine of three of four rabbits 6-13 months after intravenous or intracolonic inoculation of homogenates of rabbit mesenteric lymph-nodes after passage of human ulcerative colitis tissue (100 mum or 0.2 mum). Three of thirty A2G strain mice injected with similar tissue homogenates (100 mum or 0.2 mum) from patients with ulcerative colitis into the footpad or intraperitoneally, had granulomatous changes in footpad, bowel, liver and/or spleen 3-22 months later. Such changes did not develop in rabbits or mice inoculated with tissue from normal controls. The results of these experiments suggest that a transmissible factor is involved in the aetiology of ulcerative colitis.


A mechanism by which high-density lipoproteins may slow the atherogenic process. There is a well-documented negative correlation between plasma concentrations of high-density lipoproteins (H.D.L.) and risk of clinically evident atherosclerosis. This may relate to the postulated role of H.D.L. in the transport of cholesterol out of cells. Studies of the metabolism of lipoproteins by arterial smooth-muscle cells suggest that a second mechanism also operates, H.D.L. binds to the surface of porcine arterial smooth-muscle cells as effectively as low-density lipoprotein (L.D.L.) but is internalised and degraded much more slowly. When incubated with L.D.L. these cells show a net increment in cholesterol content. However, cells incubated with equal or higher concentration of H.D.L. under comparable conditions show no cholesterol accumulation. The presence of H.D.L. in the medium partially inhibits uptake and degradation of L.D.L. and, most important, also partially suppresses the net increment in cell sterol content induced by L.D.L. The demonstrated interaction of H.D.L. and L.D.L. could be a second mechanism contributing to the apparent protective effect of high plasma-H.D.L. concentrations in relation to atherogenesis.