Defective tumoricidal capacity of macrophages from A/J mice. I. Characterization of the macrophage cytotoxic defect after in vivo and in vitro activation stimuli.

Macrophages from A/J mice fail to develop tumoricidal activity after any of several in vivo or in vitro treatments that activate cells from C3H/HeN mice. Peritoneal macrophages from A/J mice treated i.p. with viable Mycobacterium bovis, strain BCG, killed Corynebacterium parvum, or pyran copolymer fail to develop in vitro tumoricidal activity; varying the numbers of macrophages from treated mice added to target cells, or the dose and time of treatment, or the treatment schedule of these in vivo activation stimuli did not evoke cytotoxic activity. Moreover, cytotoxic activity by macrophages from A/J mice was not observed with any of four target cell lines derived from three different mouse strains.  In vitro treatment of peritoneal exudate macrophages from A/J mice with lymphokine-rich supernatants, bacterial endotoxins, or T cell mitogens was also ineffective; varying the numbers of treated macrophages added to target cells, the dose of in vitro activation stimuli, or the time of treatment did not evoke cytotoxic activity. Thus, A/J mice exhibit a profound defect in macrophage tumoricidal capacity to both in vivo and in vitro activation stimuli over a wide range of experimental conditions.


Immune response to levan. II. T independence of suppression of cross-reactive idiotypes by anti-idiotype antibodies.

Pretreatment of BALB/c mice with antisera to a cross-reactive idiotype (E109IdX) expressed on many anti-bacterial levan (BL) and anti-inulin (Inu) antibodies leads to a prolonged suppression in production of IdX-bearing molecules in response to BL immunization. There is a comparable suppression in numbers of plaque-forming cells secreting IdX-bearing anti-BL and anti-Inu molecules. Furthermore, spleen cells from anti-E109IdX pretreated mice are unable to transfer to irradiated recipients the ability to produce IdX-bearing anti-BL and anti-Inu antibodies. These results indicate that the suppressive effect is at the precursor level and not simply a clearance of antibodies bearing the IdX. Suppression of IdX production can be achieved by pretreating nu/nu BALB/c mice with anti-E109IdX antibodies. Furthermore, spleen cells from pretreated mice do not inhibit the capacity of spleen cells from normal mice transferred to irradiated recipients to produce E109IdX in response to BL. This indicates that the suppression of IdX production in the anti-BL system is T independent and probably represents direct inhibition of precursors by anti-IdX.


In vitro killing of schistosomula of Schistosoma mansoni by BCG and C. parvum-activated macrophages.

Resistance to Schistosoma mansoni infection in the mouse has been induced either specifically by a primary infection with this parasite or nonspecifically by a variety of immunostimulants such as BCG. In the present study we developed an in vitro system to examine the effector mechanism of nonspecifically induced resistance. Activated macrophage monolayers obtained from BCG- or Corynebacterium parvum treated mice killed a respective mean 32 +/- 6% and 48 +/- 5% of schistosomula after 24 hr incubation. The killing of the parasites was verified by their inability to mature to adult worms upon injection into normal mice. The activated macrophage-mediated killing was related to cell:parasite ratio, and was partially lost if the macrophage monolayers were kept in cultures for 24 hr before incubation with the organism. Supernatants of macrophages cultured in the presence of schistosomula killed a mean of 51 +/- 3% of the organisms whereas those from cells cultured alone resulted in a mean killing of 25 +/- 3%. Furthermore, toxic supernatants could be generated equally well on incubation with S. mansoni schistosomula or Trichinella spiralis larvae. Our data show that activated macrophage monolayers through soluble mediators destroy a significant proportion of the multicellular parasite S. mansoni schistosomula in vitro.


Immunoregulatory circuits among T cell sets: effect of mode of immunization on determining which Lyl T cell sets will be activated.

Lyl T cells are able to induce B cells to make antibody and also to induce a resting Lyl23 T cell set to exert potent feedback suppression. Which of these two pathways Lyl T cells take can be influenced by the mode of immunization. In particular, in vitro immunized Lyl T cells are more likely to induce suppression than are Lyl T cells immunized in vivo even when both populations deliver the same amount of help to purified B cells. The feedback suppression induced by the Lyl T cells immunized in vitro can be distinguished from suppression mediated by Ly2+ T cells; in the former case suppression is preceded by a precocious antibody response, whereas suppression mediated by Ly2+ T cells is apparent throughout the entire period of observation.


Carrier-specific immune memory to a thymus-independent antigen in congenitally athymic mice.

Immune memory to the DNP epitope coupled to a nonmitogenic thymus-independent carrier, Ficoll, was demonstrated in congenitally athymic outbred Swiss mice. Strong IgM and modest IgG components of this memory were detected. Moreover, this memory was carrier specific since it was elicitable only when DNP-Ficoll primed mice were challenged with DNP-Ficoll and not when similarly primed mice were challenged with DNP coupled to pneumococcal polysaccharide or to keyhole limpet hemocyanin. Ficoll primed mice also demonstrated a memory response when challenged with DNP-Ficoll. These findings indicate that a non-T cell, presumably a B cell, is capable of recognizing the carrier epitopes of this thymus-independent hapten-carrier complex. Unlike their athymic counterparts, euthymic mice of the same genetic background failed to demonstrate the IgM component of this memory, but they did demonstrate modest carrier-specific IgG memory. These results strongly suggest that suppressor T cells are either directly or indirectly important in regulating the IgM memory response of these mice to DNP-Ficoll. Indirect regulation could possibly occur via an antibody-mediated specific immune suppression mechanism.


Antibody penetration into living cells. II. Anti-ribonucleoprotein IgG penetrates into Tgamma lymphocytes causing their deletion and the abrogation of suppressor function.

We have previously shown that an anti-ribonucleoprotein (RNP) IgG can penetrate into live human mononuclear cells (MNC) having receptors for the Fc portion of IgG. Because T cells with such receptors (Tgamma cells) seem to behave as suppressor cells in immune regulation and because this suppressor function is diminished in diseases where antinuclear antibodies appear, we considered the possibility that antinuclear IgG antibody could penetrate Tgamma cells and affect them. Herein we show that fluorescein-labeled anti-RNP IgG can penetrate into Tgamma cells, enriched by either mitogenic stimulation or separation with a subpopulation of T cells with low affinity for sheep erythrocytes. Incubation of MNC with anti-RNP IgG before carrying out the separation procedures resulted in apparent loss of Tgamma cells at the end of separation. To confirm that deletion had actually occurred, we performed a cytotoxicity assay using 51Cr-labeled T cells. Anti-RNP IgG had a significantly higher cytotoxic effect that normal IgG on T cells, particularly on those with low affinity for sheep erythrocytes that include most Tgamma cells. Suppressor cell function studied in a system where it was expanded, by either 7-day culture or incubation with concanavalin A, and detected in a reverse plaque-forming cell assay with rabbit anti-human immunoglobulin-developing antibody was found to be abrogated by the addition of anti-RNP IgG to the suppressor function-expanding cultures. Controls in Ig-free medium, or medium supplemented with normal human IgG, aggregated normal human IgG, BSA-anti-BSA immune complexes, or F(ab')2 fragments of the anti-RNP IgG, did not abrogate suppressor cell function. This indicates that the abrogation of suppressor cell function by anti-RNP IgG is due to its penetration into Tgamma cells. Suppressor cell loss and/or dysfunction caused by penetration of antinuclear antibodies into Tgamma cells may lead to the self-perpetuation of autoimmune disease.


Production and characterization of DR xenoantisera: use for detection of serum DR antigens.

Immunoadsorbents consisting of rabbit anti-DR antibodies bound to Staphylococcus aureus were used to bind DR antigens from NP40 cellular extracts of cultured B lymphoid cells. When injected into rabbits, these DR antibody absorbents elicited the production of antibodies that were shown by both serologic and immunochemical techniques to react specifically with the DR antigen molecule. A rosette inhibition assay with B lymphoid cells and anti-DR xenoantisera was used to detect DR antigens in human serum. DR serum antigens could be found in an enriched preparation of serum lipoproteins obtained by centrifugation in KBr. In addition, sera from some patients with neoplastic diseases contained higher levels of DR serum antigens than those found in normal individuals.


Slow reacting substance (SRS) from ionophore A23187-stimulated peritoneal mast cells of the normal rat. II. Evidence for a precursor role of arachidonic acid and further purification.

The generation of slow reacting substance (SRS) from ionophore A23187-stimulated rat peritoneal mast cells was enhanced by arachidonic acid (AA). This SRS generation was inhibited by 5,8,11,14-eicosatetraynoic acid (ETYA), an acetylenic analogue of AA and an inhibitor of both fatty acid cyclooxygenase and lipoxygenase. Indomethacin, a fatty acid cyclooxgenase inhibitor, had an enhancing effect upon SRS generation. This suggests SRS generation occurred through an ETYA sensitive step--perhaps a lipoxygenase. Radiolabel from [14C]-AA was incorporated into SRS with comigration of radioactivity and bioreactivity in silicic acid and thin layer chromatographies. Upon silicic acid chromatography, the active principle was eluted in the methanol fraction. Two-dimensional thin layer chromatography revealed chromatographic separation from other known spasmogenic substances and phospholipids. Mast cell SRS was found to display physiochemical properties similar to those of rat basophilic leukemia cell SRS, namely: that mast cell SRS generation was 1) enhanced by arachidonic acid; 2) inhibited by ETYA but not by indomethacin; 3) incorporation of [14C]-AA into the active principle; and 4) similar behavior during purification in silicic acid and thin layer chromatographies.


IgM rheumatoid factors in mice injected with bacterial lipopolysaccharides.

Bacterial lipopolysaccharides (LPS) induced the formation of IgM rheumatoid factors (RF) in several strains of mice including athymic C57BL/6 nude mice, but not in the LPS-resistant C3H/HeJ mice. The RF induced by LPS reacted not only with murine IgG but also with IgG from cows, goats, guinea pigs, and humans. The kinetics of this RF response to injection of LPS were similar to those of antibody response against DNA and a hapten, dinitrophenyl (DNP), and to those of total IgM production. In addition, the RF activity of individual serum samples correlated significantly with levels of anti-DNA and anti-DNP antibodies and of IgM. Therefore, it is concluded that the induction of RF results from polyclonal antibody synthesis by B cells stimulated with LPS. This observation suggests that LPS or LPS-like substances may help to generate RF in patients with rheumatoid arthritis or with some infectious diseases.


Circulating and tissue-bound immune complex formation in murine malaria.

Immune complex formation during Plasmodium berghei infection of OF1 mice was investigated. Circulating immune complexes (CIC) were detected by the Clg-binding assay and the conglutinin-binding solid-phase assay in lethal or drug-limited infections. CIC appeared on day 9 of infection, peaked on day 11, and disappeared only after complete cure of the infection. Analysis of the immune complexes detected by the Clq-binding assay revealed the following characteristics: sedimentation coefficients of 13S to 21S, resistance to DNAse, and selective removal by filtration through protein A bound to Sepharose. Glomerular deposits of IgM preceded the appearance of CIC, whereas deposits of IgG and C3 were concomitant with the appearance of CIC. Tissue-bound immunoglobulins were also found in the choroid plexus. The appearance of anti-malarial antibodies and malarial antigens in the serum was closely associated with a depression of C3 levels and the presence of CIC. Drug treatment was followed by normalization of C3 levels, and clearance of both CIC and malarial antigens.


Shared idiotypes of human peripheral blood B and T lymphocytes.

In a patient with an IgG lambda monoclonal serum component possessing anti-streptolysin O activity, we have demonstrated peripheral blood B and T lymphocytes with shared or similar idiotypes. The idiotypic T lymphocyte membrane structure was capable of binding the specific antigen (SLO). After radioiodination and subsequent detergent solubilization of the same T cell population, immunoprecipitation of the lysate by employing anti-idiotypic antibodies, resulted in the isolation of a polypeptide chain with a m.w. of 70,000 on SDS polyacrylamide gels under reducing conditions. The polypeptide expressed no isotypic immunoglobulin markers. Internal labeling experiments indicated that this membrane structure was actively synthesized by the T lymphocytes.


Expression of MHC antigens by mouse thymic dendritic cells.

Thymic epithelial cells express MHC antigens in several different patterns. I-A is present throughout the thymic cortex on dendritic cells. The remainder of the I region and H-2K/D are expressed on dendritic cells apparently only variably in the cortex (at least in some haplotypes). All MHC antigens tested are present in the medulla on epithelial cells; expression on medullary lymphocytes cannot be evaluated. Monoclonal anti-MHC antibodies confirm these results. The significance of these findings to T cell maturation is discussed.


Natural killer cells, bone, and the bone marrow: studies in estrogen-treated mice and in congenitally osteopetrotic (mi/mi) mice.

Mice lose natural killer cells after 6 weeks of treatment with 17 beta-estradiol. We here demonstrate that the same protocol leads to loss of genetic resistance to bone marrow transplantation and to significant osteoproliferation with loss of bone marrow. We also show that mice with reduced marrow because of congenital osteopetrosis are deficient in natural killing. These findings are consistent with previous evidence that natural killing and genetic resistance to bone marrow transplantation are dependent upon the marrow. Temporal studies of bone histology and radiology during and after treatment with estrogen reveal that alterations in natural killing proceed more rapidly than changes in bone marrow volume. These studies also demonstrate that estrogens induce osteoproliferation only at endosteal surfaces that are adjacent to hematopoietic marrow. From these observations, we conclude that estrogens do not reduce natural killer cells simply by reducing the volume of bone marrow. Estrogens may instead have an effect on bone marrow. Estrogens may instead have an effect on bone marrow cells that leads both to osteoproliferation and to a deficiency of marrow-dependent cells.


T cell regulation of polyclonal B cell responsiveness. I. Helper effects of T cells.

Polyclonal activation of murine splenic B lymphocytes to secrete immunoglobulin was shown to be subject to regulation by splenic T cells. By admixture of separated B and T cell populations it was demonstrated that normal fresh splenic T cells were able to augment polyclonal B cell responsiveness to LPS up to several-fold. Optimal collaboration between these two cell types ensued when they were co-cultured in equal numbers. T cell-mediated enhancement of polyclonal B cell responses was dependent upon the ability of T cells to divide and was manifested upon T cell interaction with B cells soon after culture initiation. Originally expounded as a one-signal phenomenon, polyclonal activation of lymphocytes by LPS is, under the circumstances described, attributable instead to two distinct, nonspecific signals acting in concert. The observation that T cells from LPS-nonresponder (C3H/HeJ) mice were deficient in the capacity to enhance polyclonal B cell responsiveness of B cells derived from responder (C3H/HeN) mice implied a direct action of LPS on the involved T cells as well as an active role for the T cell signal in this immunoregulatory event. The novel observation of a functional T cell defect in LPS responsiveness in the C3H/HeJ mouse is discussed in terms of its other cellular defects.


Evidence that natural cytotoxicity and antibody-dependent cellular cytotoxicity are mediated in humans by the same effector cell populations.

The present study strongly suggests that, in humans, natural killer (NK) activity and antibody-dependent cell-mediated cytotoxicity (ADCC) are mediated by the same effector cell population. This is supported by two different experimental approaches. First, competition for NK effector cells was accompanied by simultaneous inhibition of ADCC activity. Target cells sensitive to NK activity were capable of inhibiting specifically an ADCC assay in cold target competition experiments. Second, specific removal of NK cells on monolayers formed by target cells sensitive to NK activity caused simultaneous depletion of ADCC effector cells. In association with the removal on the monolayers of effector cells for ADCC as well as NK activity, we also found a significant depletion of cells bearing Fc gamma receptors.


Allogeneic marrow transplantation after total lymphoid irradiation (TLI): effect of dose/fraction, thymic irradiation, delayed marrow infusion, and presensitization.

BALB/c mice infused with 30 x 10(6) C57BL/Ka bone marrow (BM) cells 1 day after treatment with fractionated total lymphoid irradiation (TLI) (17 fractions of 200 rads each) became stable mixed chimeras without clinical graft-vs-host disease (GVHD). Mice given 18 fractions of 100, 50, or 25 rads each followed 1 day later by C57BL/Ka BM did not become chimeric, indicating that a critical cumulative radiation dose is required for this effect. Animals given TLI with lead shielding placed over the thymus also developed stable chimerism without GVHD. Thus susceptibility to tolerance induction and protection from GVHD after TLI and allogeneic BM transplantation is not due to alteration of the thymic microenvironment by fractionated irradiation. A delay of 7 or 21 days between completion of TLI and BM administration resulted in a high incidence of graft rejection. Sensitization to minor histocompatibility antigens of the BM donor strain by blood transfusion either before or during TLI resulted in marrow graft rejection in a high percentage of animals.


Phenotypic and functional distinctions between the TH2+ and JRA+ T cell subsets in man.

Prior work has demonstrated the existence of distinct human peripheral blood T cell subsets by utilizing heterologous as well as autoimmune antisera. In the present study, the relationship between the TH2+ and JRA+ T cell subsets was examined. T cells were purified with Sephadex G-200 anti-F(ab)2' affinity chromatography and E-rosetting technique, and subsequently fractionated into TH2+ and TH2- subsets by utilizing indirect immunofluorescence on FACS. Approximately 40 to 45% of the TH2- subset was shown to be JRA+, whereas less than 5% of the TH2+ subset was JRA+. In reciprocal studies, T cells were fractionated into JRA+ and JRA- subsets and reacted with heterologous antisera with anti-TH2+ specificity and indirect immunofluorescence. FACS analysis demonstrated that the JRA+ population contained no TH2+ T cells. In contrast, the JRA- population contained TH2+ T cells and accounted for the entire TH2+ subset found in the unfractionated T cell population. Functional studies showed that the TH2+ subset, and not the JRA+ subset, contain the effector population for cell-mediated lympholysis. It is concluded that the TH2+ and JRA+ T cell subsets define distinct and different T cell populations in man.


A comparison of methods for obtaining high yields of pure immunoglobulin from severely haemolysed plasma.

Ammonium sulphate, DEAE ion exchange chromatography, and polyethylene glycol (PEG) were compared for their ability to isolate and purify large quantities of immunoglobulins from severely haemolysed plasma. PEG, which produced the highest yield and purity, proved to be the only method which completely removed the haemoglobin.


A quantitative assay for low levels of IgM by solid-phase immunofluorescence.

A quantitative immunofluorescent assay capable of detecting between 15 and 1450 ng of IgM on a strictly linear standard curve has been developed. The assay system is unique in that in vitro biosynthesized polyclonal IgM in culture supernatants can be quantitated with a minimal quantity of supernatant. Interference created by lectins and bead-surface interactions have been analyzed. This easily operated assay has been validated and is reliable. A very important feature of the system is the considerable cost reduction in comparsion with radiometric techniques.


A microculture system for generating haemolytic antibody responses from human tonsillar lymphocytes.

Small numbers of Ficoll-Hypaque purified human tonsillar lymphocytes were stimulated with PWM to produce SRBC-specific PFC in a microculture system. The magnitude of the response varied among different tonsils but was typically between 200 and 1000 PFC/10(6) cells cultured. Little or no response was observed in the absence of PWM. SRBC failed to stimulate a SRBC-specific response and the presence of this antigen in PWM-stimulated cultures depressed the response. The time of the maximum response was inversely related to the number of cells cultured. In addition, the duration of the response was limited by rapid depletion of critical medium requirements and/or build up of inhibitory factors especially when the cell concentration exceeded 5 x 10(5) cells/culture. This effect could be partially overcome by daily feeding of cultures with fresh medium. Fractionation studies indicated a requirement for both T and B cell populations. Constant efficiency of PFC production with respect to cell number could be achieved by the addition of inactivated autologous 'filler' cells. The significance of these results and applicability of the microculture system to a detailed analysis of human antibody responses will be discussed.


An autoradiographic microtechnique to study lymphocyte stimulation in solid cultures.

A microtechnique to study lymphocyte stimulation in solid cultures by autoradiography is described. Using this technique at a concentration of 2 x 10(6) lymphocytes/chamber, it was found that the percentage of proliferating cells was 8.4% with anti-lymphocyte serum, 3.9% with calcium ionophore A23187, 2.5% with phytohemagglutinin P, 5.8% with phytohemagglutinin HA17 and 4.2% with tetradecanoyl-phorbolacetate. Lymphocyte responses were dependent on cell concentration. Lymphocyte responses to phytohemagglutinins P and HA17 were potentiated by incorporation into the cultures of 5% autologous erythrocytes.


The bovine lymphoid system: binding and stimulation of peripheral blood lymphocytes by lectins.

We tested ability of 11 different lectins to stimulate DNA synthesis in bovine peripheral blood lymphocytes, using a wide range of experimental conditions. Five of the 6 lectins which induced DNA synthesis (concanavalin A, succinyl-concanavalin A, phytohemagglutinin M, pokeweed mitogens and lipopolysaccharide) did so under conditions similar to those optimal for stimulation of murine lymphocytes. The other lectin (peanut agglutinin) stimulated normal bovine lymphocytes whereas it does not stimulate normal mouse, rat, guinea pig or human lymphocytes. The binding of 5 different lectins to bovine peripheral blood lymphocytes was measured by fluorescence microscopy. Four of the lectins bound to various proportions of cells. Double labeling experiments using a rhodamine-labeled goat anti-bovine immunoglobulin reagent and fluorescein-labeled lectins showed that both peanut agglutinin and soybean agglutinin bound to lymphocyte populations which were negative for surface immunoglobulin. The majority of lymphocytes negative for surface immunoglobulin bound peanut agglutinin, indicating that this lectin may bind specifically to bovine 'T' lymphocytes.


A sensitive hemolytic assay for mouse C3.

We have developed a sensitive hemolytic assay for mouse C3, utilizing readily available guinea pig complement components and standard techniques, which allows C3 to be detected in mouse serum diluted many thousand-fold. Mouse serum also contains a potent, relatively heat-stable C3-inactivator system, which is largely blocked by Suramin.


The ultrastructural localization of IgA deposits in chronic bullous disease of childhood (CBDC).

A case of bullous disease in a child with linear IgA immune deposits at the basement membrane zone and with some clinical, histological, and electron microscopic characteristics both of dermatitis herpetiformis and bullous pemphigoid, is described. The bulla formed between the basal lamina and basal cell membranes as in bullous pemphigoid, but at the same time there were numerous inflammatory cells in the dermis just below the partly destroyed basal lamina and also abundant fibrin deposits in very recent bulla and in the skin, all of which is rather characteristic of dermatitis herpetiformis. Ultrastructurally, the IgA deposits were located chiefly below the lamina basalis (the dermal type) but also, though less abundantly, in the lamina lucida, very much as we have seen them to be in adult cases with linear IgA immune deposits at the basement membrane zone. The investigations have supplied further evidence showing the chronic bullous disease of childhood to be actually a counterpart of the form in adults with the same linear localization of IgA deposits.


Comparison of different epithelial substrates useful for indirect immunofluorescence testing of sera from patients with active pemphigus.

A positive statistical correlation between intercellular antibody titer and disease severity was found for a group of 6 patients with active pemphigus vulgaris, irrespective of whether indirect IF testing was performed on 5 different epithelial substrates which included guinea pig esophagus, human foreskin, rabbit esophagus, monkey esophagus and monkey lip. In a group of 4 patients with pemphigus foliaceus a positive correlation between disease severity and titer was found only when rabbit esophagus was used for indirect fluorescence testing. In both groups of patients there were individual patients in whom the antibody titer was positive at a time when no disease was present and conversely, there were some patients in whom the antibody titer was negative at a time when extensive disease was present. Because of this inconsistency, the use of antibody titers to monitor disease activity and therapy in individual patients may not be justifiable.


Exfoliative cytological procedures as a nonintrusive method for dermatogerontological studies.

Exfoliative cytological procedures, based on the sticky slide and detergent scrub techniques, were used in the nonintrusive collection of horny cells from the skin surface of volunteers 20 to 30 yr of age and 65+ yr of age. These samples were analyzed with a variety of instruments, including a Magiscan image analyzer, a Vickers M-85 microspectrophotometer, and a Coulter electronic cell counter. The results indicate that age-associated changes in horny cells may be useful in monitoring objectively aging of skin and in assessing the impact of various environmental insults on aging.


Demonstration of antigenemia by radioimmunoassay in rabbits experimentally infected with Aspergillus.

Antigens were detected in the blood of rabbits infected with Aspergillus fumigatus by a solid-phase (tube) radioimmunoassay (RIA). The radiolabel for the assay was a polysaccharide-rich alkali extract (APAE) from the mycelia of A. fumigatus. Before this extract could be suitable labeled with 125I, it had to be conjugated with tyramine. Rabbits immunized with heat-killed mycelia had titers of antibody in serum of as high as 1:38,000 against this radiolabeled antigen. With unlabeled and unconjugated APAE as the standard antigen, the sensitivity of the RIA was 12 ng per test, or 500 ng/ml. Antigenemia was detectable by RIA three days after infection of rabbits with A. fumigatus. Blood cultures taken concomitantly were uniformly negative. These results indicate that antigenemia occurs in invasive aspergillus infection and in such cases can be detected by RIA. These observations may be important in the diagnosis of invasive aspergillus infections in humans.


Prevention of colonization of the urinary tract of mice with Escherichia coli by blocking of bacterial adherence with methyl alpha-D-mannopyranoside.

Methyl alpha-D-mannopyranoside (alpha MM), a competitor inhibitor of the binding of mannose by Escherichia coli, was tested for its ability to prevent infection of the urinary tract of mice with infective strains of the organisms. Injection of the bacteria in the presence of the drug resulted in a considerable reduction in the number of bacteriuric mice. In this system alpha MM was inactive against Proteus mirabilis in accordance with its inability to inhibit the adherence of this organism to epithelial cells in vitro, and methyl alpha-D-glucopyranoside proved inactive against both E. coli and P. mirabilis.


Experimental viral infections of the temporal bone. I. Experimental otitis media due to vaccinia virus in hamsters.

Clinical and pathologic studies of patients with smallpox suggest that conductive hearing loss is a frequent complication of smallpox and that it is due to otitis media. A possible experimental model for this hearing loss has been developed by study of the pathogenesis of acute vaccinia virus infections of hamster middle ears. Infant hamsters developed severe necrotic otitis media, and vaccinia viral antigen was demonstrated by fluorescent antibody staining in large confluent areas of epithelial cells lining the middle ear. Juvenile hamsters developed a patchy, less severe infection of the middle ear, most of which present along the tympanic membrane and middle ear ossicles, Adult hamsters did not develop middle ear infection. These findings support the argument that variola (smallpox) virus can cause otitis media in humans and that viral infection of the middle ear may be the cause of hearing loss.


The incidence of Salmonella in random-source cats purchased for use in research.

In research facilities, cats are routinely ignored as a potential source of salmonella infection. Over a period of 18 months, 142 cats received from commercial vendors for use in research were screened for enteric Salmonella. Salmonella was isolated from 15 animals, an incidence of 10.6%. Five (29%) of the 17 shipments contained animals that were positive for Salmonella. The serotypes isolated were Salmonella derby, Salmonella typhimurium, Salmonella anatum, Salmonella enteritidis, and Salmonella bredeney.


A nosocomial outbreak of infections due to multiply resistant Proteus mirabilis: role of intestinal colonization as a major reservoir.

An outbreak of nosocomial infections involving an unusual strain of multiply resistant Proteus mirabilis (phage type 8888) occurred in 15 patients, 14 of whom were in the surgical intensive care unit at that time. No common source of infection was identified, and person-to-person transmission was the most likely mode of spread. Case-control analysis indicated a significantly increased risk infection related to length of hospital stay (P less than 0.005), number of operations (P less than 0.005), proximity to another case (P less than 0.01), number of antibiotics received (P less than 0.02), and use of a respirator (P less than 0.01). Only the number of operations (P less than 0.01) and proximity to another case (P less than 0.05) remained significant risk factors when related parameters were controlled by multivariate analysis. Thirteen of 14 patients prospectively cultured were colonized by the epidemic organism in the intestinal tract, while rectal carriage preceded infection by the same strain in at least four patients. These data suggest that intestinal colonization may have been an important reservoir for this outbreak, and the findings may explain the unduly prolonged course of intrahospital spread as well as the difficulty encountered in the eradication and control of the outbreak.


[Investigation of Pd-Ag alloy for porcelain fusing. (Part 3) Mechanical properties (author's transl)].

The most suitable composition of Pd-Ag alloys added In and/or Sn for porcelain fusing was determined by means of the measurement of the mechanical properties (tensile strength, elongation and hardness). The results were summarized as follows: (1) This highest tensile strength was obtained when 5% In and 5% Sn, or 10% Sn were added to the 60 Pd-40 Ag mother alloy, and it was about 52 kgf/mm2 (570 MPa). (2) The effect of the addition of In and Sn to the tensile strength depended on the concentration of Pd and Ag. (3) The elongation increased with increase of the amount of Pd. (4) The Vickers hardness number of the alloys was ranged between 100 and 200. It increased with increase of Ag or Sn content. (5)  The most suitable compositions of Pd-Ag alloys for porcelain fusing deduced from the properties such as the bonding strength (in part 1), the thermal expansion coefficient (in part 2) and the mechanical properties (in this paper) were 60 Pd-40 Ag or 64 Pd-36 Ag with addition of 4 Sn-6 In or 8 Sn-2 In. Some characteristic properties of these alloys were as follows; the bonding strength 220 kgf/cm2 (22 MPa), thermal expansion coefficient 14.7 +/- 1x10(-6)/degrees C, thermal expansion hysteresis. 0.00 approximately 0.02%, tensile strength 51 kgf/mm2 (500 MPa), elongation 5% and Vickers hardness number 160 approximately 190.


[Studies on Ni-Ta alloys. (Part 2) Age-hardening and metal-to-ceramic bonding (author's transl)].

Age-hardening and metal-to-ceramic bonding were studied on Ni-Ta alloy for dental application. In the first experiment, it was shown the precipitation-hardening was recognized in Ni-33 wt% Ta, Ni-34 wt% Ta except for Ni-30 wt% Ta. The precipitates were of a Widmamstätten type structure and appeared to be a intermetallic compound, namely Ni3Ta, from the results of X-ray diffraction (debyesherrer) despite of the close proximity of the diffraction patterns of Ni and Ni2Ta. In the second experiment, it was shown that the bonding strength was 176 kg/cm2 for Ni-30 wt% Ta-ceramic system, compared with that of 222 kg/cm2 of Wiron S-ceramic system. The oxide layer was as wide as 3 approximately 5 micron at the Ni-30 wt% Ta-ceramic interface, in which high concentrations of Ni, Ta, and low concentrations of Al, Si were observed. The coefficient of thermal expansion, which was larger than that of porcelain, was 17.4 x 10(-6)/degrees C for Ni-30 wt% Ta.


Sorting out behaviour of disaggregated cells in the absence of morphogenesis in Dictyostelium discoideum.

Cells disaggregated from slugs of Dictyostelium discoideum were cultured in Bonner's salt solution in roller tubes. Cells rapidly stuck together to form an amorphous loose agglutinate which was later transformed into a spheroidal tight agglutinate surrounded by slime sheath material. Prespore cells in the loose agglutinate underwent partial dedifferentiation by starting to decompose their specific antigen until formation of the tight agglutinate, in which the antigen was resynthesized. During the process, there was some decrease in the proportion of prespore cells. Changes in the distribution of prespore and prestalk cells in the agglutinates were examined by using immunocytochemical staining. They were randomly distributed in the early agglutinates, but became well separated in 4 h agglutinates in such a way that prestalk cells were completely enveloped by prespore cells. Prestalk cells later came outside to be partially enveloped and finally occupied a hemisphere side by side with prespore cells. During the process, cells in one or two outer layers differentiated into prestalk or stalk cells. Similar changes in the distribution pattern were observed, when labelled prestalk cells were cultured with unlabelled prespore cells and their distribution in co-agglutinates was followed by autoradiography. It was concluded from these results that the majorities of prestalk and prespore cells isolated from slugs are sorted out in agglutinates without changing their original cell types, and that the sorting-out occurs in the complete absence of polar structures and movement of the cell mass. The distribution patterns in agglutinates of prestalk and prespore cells were discussed with reference to intercellular adhesion among/between them.


Transfer of specific unresponsiveness to organ allografts by thymocytes. Specific unresponsiveness by thymocyte transfer.

Prolonged survival of vascularized organ allografts has been produced in unmodified inbred rats by transfer of thymocytes from enhanced, engrafted, syngeneic animals. For these thymocytes to increase significantly the survival of test allografts they must be harvested 6-9 d after transplantation. Thymectomy of the enhanced, engrafted animals during the same critical period causes acute rejection of othewise long surviving grafts. For optimal effect, the enhanced thymocyte donor must be actively and passively immunized and receive a cardiac allograft. The necessity for erythrocytes in the initial active immunization regimen is noted. Additionally, the antigenic specificity of the suppressor effect has been established with two histoincompatible donor rat strains. Cellular and humoral host responses mounted by test graft recipients after thymocyte transfer from enhanced, engrafted donors are different from those mounted either by unmodifed animals acutely rejecting their grafts or by enhanced rats bearing well-functioning grafts. Numbers of T lymphocytes are reduced in the grafted hearts and in the spleens of test graft recipients, a finding paralleled by the complete absence of specific direct lymphocyte-mediated cytotoxicity. In contrast, cytotoxic antibody production, although delayed, is increased in magnitude, peaking around the time of graft rejection. These studies provide evidence that different biological manipulations can modify separate pathways in the complex cellular and humoral responses towards organ allografts. They demonstrate that cellular immunity is critically involved in immunological enhancement of vascularized organ allografts, a phenomenon hitherto considered primarily humoral. It seems clear that cells with suppressor activity are present within the thymus during the early phases of immunological enhancement.


A review of the metabolism of the aspartyl moiety of aspartame in experimental animals and man.

Aspartame (3-amino-N-(alpha-carboxyphenethyl) succinamic acid, methyl ester; the methyl ester of aspartylphenylalanine, SC-18862) is hydrolyzed in the gut to yield aspartic acid, phenylalanine, and methanol. This review of the literature describes the metabolic paths followed by aspartate in its conversion to CO2 or its incorporation into body constituents. About 70 percent of 14C from [asp-14C]-aspartame is converted in the monkey to 14CO2. Some of the aspartate is converted at the intestinal mucosal level to alanine by decarboxylation. This amino acid may be oxidized to CO2 by entering the tricarboxylic acid cycle via pyruvate and acetyl CoA. In addition, transamination of aspartate to oxaloacetate permits this product also to enter the tricarboxylic acid cycle. Aspartate may also be incorporated into body constitutents such as other amino acids, proteins, pyrimidines, asparagine, and N-acetylaspartic acid. It is concluded that the aspartate moiety of aspartame is metabolized in a manner similar to that of dietary aspartic acid.


Activation of macrophages in vivo and in vitro. Correlation between hydrogen peroxide release and killing of Trypanosoma cruzi.

As reported previously, mouse peritoneal macrophages could be activated to kill intracellular trypomastigotes of Trypanosoma cruzi, the agent of Chagas' disease, in either of two ways: by immunizing and boosting the mice (3), or by culturing resident or inflammatory macrophages in spleen cell factor(s) (SCF) in vitro (2). Macrophages activated in vivo became less trypanocidal with time in culture, and cells activated in vitro lost trypanocidal capacity when CSF was removed (2). In the present study, the ability of macrophages to release H2O2 in response to phorbol myristate acetate (PMA) could be induced in vivo and in vitro, and reversed in vitro, in a manner correlating closely with changes in trypanocidal activity. Macrophages could be activated in vitro with SCF in a time-dependent and dose-dependent fashion, so that they released as much H2O2 as macrophages activated in vivo. The sensitivity of epimastigotes and trypomastigotes to enzymatically generated H2O2 suggested that the generation of H2O2 by activated macrophages could be plausible explanation for their trypanocidal activity. Of the biochemical correlates of macrophage activation reported to date, increased ability to release H2O2 seems most closely allied to enhanced capacity to kill an intracellular pathogen.


NCTR computer systems designed for toxicologic experimentation. III. Breeding information system.

The Breeding Information System (BIS) facilitates management control of the breeding colony operation at the National Center for Toxicological Research (NCTR). Although this automated data handling system was initially intended to support Animal Husbandry the system's basic design, flexibility or reporting, data manipulation capabilities, and integration with other NCTR data collection systems provides BIS with capabilities that have application to other groups including the Plans and Programs and most scientific areas. This description of the System is in terms of its potential value to these diverse user groups.


Purification and properties of an extracellular blastogen produced by group A streptococci.

An extracellular product of group A streptococci which induces lymphocyte blastogenesis has been purified to homogeneity by DEAE-cellulose and CM-cellulose chromatography. The protein, termed streptococcal blastogen A, has a mol wt of approximately or equal to 17,500 and is inactivated by protease treatment and by heating at 100 degrees C. The purified blastogen gave rise to multiple protein bands on nondenaturing polyacrylamide gel electrophoresis, only two of which possessed blastogenic activity. Treatment of the protein with dithiothreitol before electrophoresis resulted in the apparent conversion of the multiple forms to a single active species. Blastogen A differs in electrophoretic mobility from the streptococcal pyrogenic exotoxins and its lymphocyte stimulating activity is not inhibited by rabbit antisera to the exotoxins. An enzyme immunoassay has been developed to measure human antibodies against blastogen A. A selection of sera with varying levels of anti-DNase B contained antiblastogen A-IgG.


Basal lamina heterogeneity in the glomerular capillary tufts of human kidneys.

Two classes of glomerular basal laminas are identified with a newly developed guanidine technique. The electron-opaque epithelial basal lamina is the most prominent element of the glomerular basal lamina scaffold. It is a continuous layer within each glomerulus, folding into capillary tufts and loops, but never completely encircling the entire circumference of each capillary, similar to the serosa covering the intestinal loop and mesentery. The vascular space so defined is further partitioned into individual capillary lumen by an electron-lucent mesangial basal lamina, that forms a meshwork continuous with the vascular pole of the glomerulus and extends peripherally to surround capillary lumens. The latter, designated endothelial basal lamina, is extremely attenuated and appears as a vestigial structure in glomerular capillary loops. Changes in juxtamesangial epithelial basal lamina indicate that it may be the site of the bulk removal and renewal of the epithelial basal lamina. The unique epithelial origin of glomerular capillary basal lamina and its organization provide a structural basis for understanding the glomerular physiology gained by various tracer studies. The results also suggest that the guanidine technique may be a useful new approach to the analysis of basal lamina alterations in various glomerular diseases.


Quantitative determination of titers of anti-zona serum.

The titers of rabbit antiserum against isolated mouse zonae pellucidae were examined by several methods in connection with inhibitory effect on fertilization. The titers determined by zona precipitate, zona dissolution, indirect immunofluorescence and in vitro fertilization tests were 2(4), 2(1) to 2(4), 2(7) and 2(4), respectively. Cytotoxic effect could not be detected from zona antibody. The indirect immunofluorescence was most sensitive for detection of zona antibody but did not represent the extent of inhibitory effect on fertilization. The titer obtained by zona precipitate test was most close to the titer obtained by inhibitory effect on in vitro fertilization. The present study also demonstrated that at least 0.0375 ml of antiserum per female mouse, equivalent to 0.15-0.25% of body weight, was necessary for inhibition of fertilization in vivo by passive immunization with anti-zona serum.


The clinical phenomenology of presenile dementia. A critical review of the literature.

The more recent literature addressing the clinical phenomenology of presenile dementia is reviewed. This survey, spanning approximately the past 4 decades and comprising a large number of articles in scientific journals, reveals that reported epidemiological data, symptoms and signs, and results of ancillary tests often vary widely from study to study, especially as regards findings of history and of mental status and neurological examinations. There is considerable indication that diagnostic specificity in presenile dementia in the absence of pathological examination of the brain by tissue biopsy or at autopsy may not be as reliable as is generally thought to be the case. The observed problems with interstudy agreement are discussed and are seen to be due in large measure to methodological inconsistency and nonuniformity. Areas of difficulties in methods are identified and guidelines for subsequent, needed clinical studies are suggested.


Agreement measurement and the judgment process.

This paper reviews the rationale for correcting for chance agreement beween two raters. It is suggested that under certain conditions correction for chance agreement is both unnecessary and inappropriate. When a chance correction is indicated, the chosen measure should be one that can be shown to be logically in accord with the two judges' method of determining each case's disposition. Two agreement measures, kappa (K) and Maxwell's Random Error Coefficient of Agreement (RE), are described in common terms and are then compared in terms of their assumptions about the judgment process. The plausibility of K's assumptions are challenged, and K's use is discouraged on the grounds that judgment table data are not treated in a way compatible with the process by which the table was constructed during the judgment process itself. Use of Maxwell's RE is recommended under conditions of equal proportions in the disagreement cells.


Randomized clinical trials in the evaluation of surgical innovation.

Randomized clinical trials are widely accepted as the standard for evaluation of therapeutic innovation in many fields of medicine. The three basic components of such trials (concurrent comparison, random allocation, and objective observation) are designed to control four forms of bias (chronology bias, susceptibility bias, compliance bias, and observation bias) that may interfere with the interpretation of the results of a study. Only 2% of the articles evaluating therapeutic maneuvers published in the Journal of Neurosurgery have attempted to use concurrent controls. Only one of 863 such articles met the criteria for a randomized clinical trial. Reasons for underutilization of such trials in neurosurgery are discussed and suggestions for their wider use are offered.


Composition of isolated edema fluid in cold-induced brain edema.

Edema fluid isolated from cats with cold-induced brain edema was subjected to analysis of electrolyte content, enzyme activities, colloid osmotic pressure and the radioactivity of intravenously injected 99mTc-labeled albumin. The findings corroborate the essential features of vasogenic edema, such as its origin from the blood plasma, its rapid propagation into the white matter of the brain as contrasted with the delayed spread into gray matter, and its contribution to composition of cerebrospinal fluid. Moreover, the elevated activities of cellular enzymes and K+ content of edema fluid point to the admixture with cellular contents due to the freezing damage.


Surgical hemostasis.

Every surgical procedure taxes the hemostatic defenses of the patient. If his hemostatic mechanism is sound, he is unlikely to have a bleeding problem during or after an operation, unless, of course, a suture or clip slips off. Two classes of patients do present bleeding problems to the surgeon. One group has a pre-existing bleeding tendency, the other acquires it during or after the operation. The recognition of patients with severe hemostatic disabilities, such as hemophilia, presents no problem since the patient is aware of the disease. The mild bleeder is less likely to be detected by screening tests than by adroit questioning. The major hemostatic defect that may develop during an operation, or shortly thereafter, is disseminated intravascular coagulation. This syndrome, always secondary, may accompany shock, mismatched blood transfusion, septicemia, or extensive malignancy. Its prevention or early recongnition is much easier than treatment after circulating platelets and some coagulation factors have been consumed and fibrinolysis is destroying fibrin and fibrinogen.


Effects of zinc deficiency upon pituitary function in sexually mature and immature male rats.

Serum pituitary levels of growth hormone (GH), thyrotropin (TSH), prolactin (PRL), luteinizing hormone (LH) and follicle stimulating hormone (FSH) were measured in sexually mature (adult) and sexually immature (juvenile) male rats who had been deprived of dietary zinc for 15 and 7 weeks, respectively. When compared to pair-fed control rats receiving a zinc supplemented diet, both the adult and juvenile zinc deficient rats had significantly lower body weights, tail lengths and ventral prostate weights. The testes of the sexually immature rats were also smaller than those of the pair-fed animals. In sexually mature, zinc deficient rats serum concentrations of GH and testosterone were significantly lower and serum LH levels significantly higher than in ad libitum fed control rats. Pituitary and hypothalamic levels of other hormones did not differ from values recorded in control animals. In sexually immature zinc deficient rats serum concentrations of GH were also significantly depressed; pituitary content and concentration of LH and pituitary and serum levels of FSH were significantly increased over control values. No discernible effects of zinc deficiency upon hyplthalamic content of LH-releasing hormone or serum concentrations of PRL or TSH were recorded in juvenile rats. Zinc deficiency has minimal effects upon the hypothalamic-pituitary axis of sexually mature rats. In sexually immature males, zinc deprivation leads to impairment of gonadal growth and increased synthesis and/or secretion of the pituitary gonadotropins.


Teeth transplanted across a multiple non-H-2 barrier stimulated weak cell-mediated immunity without memory.

The genetic and immunologic criteria required for successful tooth transplantation have yet to be defined. Teeth grafted across strong barriers elicit vigorous immunity. However, teeth and tooth germs transplanted to subcutaneous sites in H-2 compatible but multiple-non-H-2 disparate recipients stimulated weak in vitro cellular cytotoxicity compared with skin grafts 5 mm in diameter. Furthermore, tooth allografts did not sensitize recipients as measured by the rejection of second-set, donor skin grafts. It may be that tooth allografts provoke fewer detrimental immune mediator systems than do skin grafts across moderate to weak transplantation barriers.


Long-term results of alveolar ridge augmentation.

Autologous onlay rib grafts in 22 patients were evaluated for bone remodeling patterns. The patients had been followed up for four to ten years; the mean was 6.2 years. The unquantitated increase in bony mass of the mandible, with altered ridge form and subsequent soft tissue surgery, provided consistent, predictable results and return to excellent denture function with long-term stability.


Neurogenic pulmonary edema in childhood.

Three patients presenting with pulmonary edema associated with head trauma and increased intracranial pressure are described. Pulmonary edema is a clearly recognized complication of head trauma; the pathogenic mechanisms appear to be regulated by increased intracerebral pressure, sympathetically induced vascular hypertension, and increased pulmonary capillary permeability. If there is evidence that neurogenic pulmonary edema is the underlying etiology, therapeutic modalities should be directed at reducing intracranial pressure and strict attention paid to the interaction between intrathoracic and intracranial pressures in order to avoid the high mortality rate associated with this condition.


Septicemia in association with acute lymphoblastic leukemia.

Fifty consecutive episodes of septicemia were studied in 41 children who had acute lymphoblastic leukemia. Seventy-six percent of these episodes occurred when the absolute granulocyte count was 200/mm3 or less and were caused by gram-negative enteric and gram-positive mucocutaneous bacteria. In eight patients, Streptococcus pyogenes was isolated at the time when ALL was diagnosed. Multiple anaerobic and aerobic isolates from a single blood culture were associated with abdominal distress, whereas Streptococcus pneumoniae and Hemophilus influenzae septicemia occurred in associated with respiratory illnesses. When patients with severe compromise of anatomic barriers or respiratory disease were excluded, 94% of all patients with septicemia had an AGC of less than 200/mm3. The data provide guidelines for treatment for febrile patients with ALL based upon the AGC, the phase of the disease, and on the presence of associated respiratory or abdominal findings.


Rapid diagnosis of type III group B streptococcal meningitis by latex particle agglutination.

A latex particle agglutination assay was developed to detect type III group B streptococcal antigen in cerebrospinal fluid. Agglutination was observed in each of 12 initial and 54% of 26 subsequent CSF specimens obtained a mean of 53 hours after the institution of antimicrobial therapy. Latex agglutination was more sensitive than countercurrent immunoelectrophoresis for the detection of type III group B streptococcal antigen in CSF from neonates and young infants with meningitis.


Renal retransplantation in children.

Evaluation of 75 cadaver donor retransplants revealed that the primary factor influencing allograft survival is patient responsiveness as reflected by sensitization with preformed cytotoxic antibodies. Actuarial allograft survival rates for nonpresensitized (less than 5%) and moderately presensitized (5 to 50%) recipients were significantly (P less than 0.01) better than those of highly presensitized (greater than 50%) recipients. Although HLA A&B antigen histocompatibility did not have a statistically significant effect on retransplant outcome, it appeared to influence allograft survival in the highly presensitized recipient. An approach to the management of children who lose an initial or subsequent allograft is indicated by these data.


Focal glomerulosclerosis and renal transplantation.

Eighteen patients with corticosteroid-resistant nephrotic syndrome developed end-stage renal disease and received one or more renal allografts. The lesion of focal segmental glomerulosclerosis and/or of focal glomerular obsolescence was demonstrable in the native kidneys of each patient. Following transplantation, nephrosis developed in three recipients. Two recipients developed nephrosis at two weeks and nine months posttransplant in association with rejection; the lesion of FGS was present in association with chronic rejection. Only one recipient developed recurrence of nephrosis and FGS unrelated to rejection. This was manifested by immediate onset of nephrosis in two successive allografts and histologic evidence of the lesion of FGS. The immediate recurrence in successive allografts suggests a circulating factor responsible for the renal lesion in this patient and indicates a separate etiology for a small number of patients with corticosteroid-resistant nephrosis and FGS.


Hemolymph of Anopheles stephensi from uninfected and Plasmodium berghei-infected mosquitoes. 2. Free amino acids.

Determinations were made of free amino acids in hemolymph collected from adult female Anopheles stephensi mosquitoes. The hemolymph first was fractionated by extraction and precipitation procedures, after which qualitative determinations of free amino acids were made by high voltage thin layer electrophoresis, and thin layer chromatography. Subsequent quantitative determinations were made with an automatic amino acid analyzer. The concentration of total free amino acids in the hemolymph rose 60--70% after the mosquito took a blood meal, and remained relatively constant thereafter. When mosquitoes took a blood meal infected with the rodent malaria parasite Plasmodium berghei, the rise in total free amino acids was only 15--25%. The chief differences that occurred with individual free amino acids was that infected mosquitoes had greater increases in arginine, greater decreases in valine and histidine, and a total loss of detectable methionine.


Association of autoantibodies with anemia, splenomegaly, and glomerulonephritis in experimental African trypanosomiasis.

Rats experimentally infected with Trypanosoma brucei rhodesiense developed a syndrome characterized by anemia, splenomegaly, and glomerulonephritis. Serologic evaluation revealed that the syndrome was accompanied by the presence of 3 autoantibodies--cold-active hemagglutinin, immunoconglutinin, and antibody to fibrinogen/fibrin products. Fluorescein isothiocyanate conjugated antibody tests showed the presence of fixed complement and fibrinogen on both trypanosomes and erythrocytes. All infected rats died by the ninth day of the infection with 5 animals showing signs of pulmonary involvement and shock. From these observations it is suggested that autoantigens, autoantibodies, and complement may have been causal in this syndrome.


In vitro development of Trichostrongylus axei, from infective larvae to young adults.

Advanced stages of Trichostrongylus axei were grown from exsheathed infective larvae in roller culture systems consisting of a complex cell-free medium, API-6, when used alone or supplemented for 8- or 21-day intervals with pepsin or reducing agents, or both. Young adult males and females were the most advanced stages attained. This stage was attained by days 18--21, when medium API-6 contained pepsin for 8 or 21 days, with or without reducing agents for the first 8 days; and by days 24--26, when medium API-6 was nonsupplemented. Trichostrongylus axei did not advance beyond early fourth stage or fourth molt when API-6 contained reducing agents for 8 or 21 days or reducing agents and pepsin for 21 days. The optimal culture system for development to young adults required medium API-6 supplemented with pepsin for 21 days and reducing agents for the first 8 days. This system produced maximal yields of less than 1% young adults in populations of about 100,000 T. axei by days 21--28.


Hemolymph of Anopheles stephensi from noninfected and Plasmodium berghei-infected mosquitoes. 3. Carbohydrates.

Determinations were made of carbohydrates in hemolymph collected from adult female mosquitoes (Anopheles stephensi). First the hemolymph was fractionated by extraction and precipitation procedures, after which qualitative and quantitative determinations of carbohydrates were made by thin layer chromatography. The most abundant sugars found in the hemolymph were glucose and trehalose, though maltose, glucuronic acid, and inositol could be found after the mosquitoes took blood meals. After the mosquitoes ingested a noninfected blood meal, their hemolymph sugar levels rose almost 4-fold. There was less of an increase following a blood meal infected with the rodent malaria parasite, Plasmodium berghei. Depletion of sugars in the hemolymph of infected mosquitoes may result from direct utilization of sugar by the malaria parasite developing within the mosquito.


Isolation of Plasmodium berghei by hemolysin lysis of infected erythrocytes and evidence for a parasite hexokinase.

A rapid and simple procedure has been developed for the isolation of Plasmodium berghei parasites from infected-mouse erythrocytes employing the heat stable hemolysin produced by Pseudomonas aeruginosa. Using parasites isolated by this method, the presence of a parasite specific hexokinase has been demonstrated, providing an explanation for the increased glucose consumption observed with infected cells. Enzyme assays and serology were employed in determining the purity and yield of purified parasites. The enzyme assays showed that about 25% of the parasites in infected RBCs were recovered in the purified state. The purified parasites were not agglutinated by rabbit-anti-mouse RBC serum which indicated the purified parasites were not contaminated by RBC components.


Ultrastructure of the rectum of infective-stage Wuchereria bancrofti (Nematoda: Filarioidea).

The authors have examined the ultrastructure of the rectum of infective-stage Wuchereria bancrofti by transmission electron microscopy. Our observations show that the rectum is divided into anterior and posterior segments. The cells of the anterior rectum appear to be derived from the microfilarial R (rectal) cells described by other authors. In both stages, these cells show voluminous nuclei, abundant mitochondria, and small cytoplasmic processes which contain fibrillar components. Amorphous material associated with these processes appears throughout the larval rectum and may protrude from the anus as the rectal plug. In the specimens examined, a patent lumen could not be traced completely through the anterior rectum. The posterior rectum has no counterpart in published accounts of microfilarial ultrastructure and probably arises during larval morphogenesis; it is lined with invaginated body cuticle, overlaid by a single layer of epithelial cells which may be of hypodermal origin.


Information, choice, and reactions to stress: a field experiment in a blood bank with laboratory analogue.

Two experiments examined the effects of various operations of personal control on reactions to stress. The first study incorporated two features into the blood drawing procedure at a blood bank: providing donors with accurate information and allowing donors to choose the arm to be used. Measurement of nurses' actions to prevent donors from fainting and self-reports of discomfort revealed that the combination of choice and information was somewhat effective in reducing distress. However, providing either information or choice alone was more effective. In a second laboratory study using a cold pressor stimulus as stressor, subjects given a choice (the option to terminate the aversive stimulus and choice of hand used) showed a reduction of aftereffects on a measure of attention to detail. Subjects given information but not choice also showed this reduction. Combining information and choice was no different from either treatment alone. Taken together, the results of both studies indicate that moderate levels of choice and information are optimal for coping with stress. An explanation was suggested based on a contextually determined relationship amomg choice, information, and perceived control.


Views on the future of veterinary education.

In briefly reviewing the history of veterinary education, particularly that in England, the enduring influence education has on the achievements and status of the profession is emphasised. The sound influence of the first teachers in South Africa, under the guidance of men like Theiler and subsequently du Toit, on veterinary education is outlined. The changes that have taken place in the face of altered demands, from that required by a predominantly State-employed profession to one consisting mainly of private practitioners but with ever-expanding fields of activity, necessitates continual attention to thorough teaching in the basic subjects, to the discipline as against the species approach--the latter being of value, however, when applying preventive medicine to herds/flocks, to a uniform undergraduate curriculum, and to foreseeing and providing adequate, specialised post-graduate education. The present under-graduate course has been extended from five to five-and-a-half years and the South African student enrollment to 90/a. Furthermore, more time will be found by already starting the teaching of Anatomy in the first year, thus providing for a final year of "student-internship". A completely newly planned faculty complex at a more favourable site is a necessity; clinical assistants will haevto be employed. More attention must be given to Public Health and Food Hygiene. Facilities for training Black studnets is a first priority in providing for additional student enrollment. Despite all modern teaching aids, the teacher must always play a vital key role.


The South African Veterinary Association: glimpses from 75 years of service to veterinary science.

The South African Veterinary Association originated with the formation of the Transvaal Veterinary Medical Association in 1903. The amalgamation of the three existing provincial associations under a joint constitution in 1920 formed a body representing the veterinary profession in the whole of South Africa. Throughout the past 75 years the SAVA has served as representative for the diversity of interests of the profession. The aims and major contributions of the Association, past and present, as well as the challenges of the future, are outlined.


[The South African Army Veterinary Services].

The history of the South African Military Veterinary Services is outlined from the time the first veterinarian, Thomas Burrowes, attached to the 8th Light Dragoons, set foot in the Cape in 1799, until the end of World War II. It summarises the extensive literature published mainly by Curson in previous issues of this Journal and extends it to include the period of that war.


Factors affecting prolapse of the prepuce in bulls.

The present state of our knowledge on the symptoms, pathogenesis and breed susceptibility of preputial prolapse in bulls is reviewed together with the anatomical and physiological relationships that operate in the development of a prolapse of the inner layer of the sheath. Suggestions for the elimination of the condition through selection are made.


The comparative antimicrobial sensitivity of Escherichia coli isolated from colisepticaemic broilers.

The standardisation of a method to perform antibiograms using Meuller Hinton agar routinely is described. It was found that from 1973 to 1975 the resistance of E. coli to oxytetracycline decreased from 49% to 32.5% and to sulphonamides from 49% to 42.5%. Resistance to furazolidone increased from 12% to 22%. No dramatic changes in the resistance to neomycin and chloramphenicol were found.


Structural requirements in positions 1, 2, 3, and 6 of the luteinizing hormone-releasing hormone (LH-RH) for antiovulatory activity.

Sixteen analogues of the luteinizing hormone-releasing hormone (LH-RH) were synthesized by the solid-phase method. In new and surprising relationships, it was found that the substitution of D-Trp into position 3 of [D- less than Glu1,D-Phe2,amino acid3,D-Phe6]-LH-RH significantly enhanced the antiovulatory potency, but substitution by Pro, N-Me-Phe,N-Me-Leu, or L-Trp reduced antiovulatory activity. The substitution of L- less than Glu in position 1 of [D-Phe2,Pro3,D-Phe6]-LH-RH by cyclohexylcarbonyl (Chc), benzoyl (Bz), Ac, Hyp, Ac-Met, hydrogen, Pro, and D- less than Glu residues, and the substitution of D-Phe in position 2 by D-Trp, D-His, D-Phg, and L-Phe residues resulted in analogues with no antiovulatory activity at 750 microgram/rat. Structural requirements for the design of inhibitors of higher potency have been discussed.


2-Acetylpyridine thiosemicarbazones. 1. A new class of potential antimalarial agents.

Based on the antimalarial properties observed for 2-acetylpyridine 4-phenyl-3-thiosemicarbazone (1), an extensive series of related thiosemicarbazones was prepared and tested against Plasmodium berghei in mice. Screening results indicated that the presence of the 2-pyridylethylidene group was critical and that certain phenyl, benzyl, phenethyl, or cycloalkyl groups at N4 of the thiosemicarbazone moiety also contribute to antimalarial activity.


Dipeptides of O-methyl-L-threonine as potential antimalarials.

L-Leucyl-O-methyl-L-threonine, O-methyl-L-threonyl-L-leucine, and O-methyl-L-threonyl-O-methyl-L-threonine were prepared and compared with O-methyl-L-threonine and L-leucine for antimalarial activity against Plasmodium berghei in mice. O-Methyl-L-threonine significantly prolonged survival time at doses of 160, 320, and 640 mg/kg. O-Methyl-L-threonyl-O-methyl-L-threonine was less active, significantly prolonging survival time only at 640 mg/kg. L-Leucine, as well as the other two dipeptides, exhibited no activity in this test.


Inhibition of anion permeability by amphiphilic compounds in human red cell: evidence for an interaction of niflumic acid with the band 3 protein.

In human erythrocyte, permeability to the anion is instantaneously, reversibly, and noncompetitively inhibited by the nonsteroidal anti-inflammatory drug, niflumic acid. The active form of this powerful inhibitor (I50 = 6 X 10(-7) M) is the ionic form. We demonstrated that: (i) The binding of niflumic acid to the membrane of unsealed ghosts show one saturable and one linear component over the concentration range studied. The saturable component vanishes when chloride transport is fully inhibited by covalently bound 4-acetamido-4'-isothiocyano stilbene-2,2'-disulfonic acid (SITS). Our estimate of these SITS protectable niflumate binding sites (about 9 x 10(5) per cell) agrees with the number of protein molecules per cell in band 3. These sites are half-saturated with 10(-6) M niflumic acid, a concentration very close to I50. (ii) Niflumic acid inhibits the binding reaction of SITS with anion controlling transport sites. These results indicate that niflumic acid and SITS are mutually exclusive inhibitors, suggesting that niflumic acid interacts with the protein in band 3. Niflumic acid also decreases glucose and ouabain-insensitive sodium permeabilities. However, these effects are produced at a very high concentration of niflumic acid (in millimolar range), suggesting unspecific action, possibly through lipid phase.


Water permeability of yeast cells at sub-zero temperatures.

A combined cryomicroscopic-multiple nonlinear regression analysis technique has been used to determine the water permeability of the yeast cell Saccharomyces cerevisiae during freezing. The time rate of change in volume of "supercooled" yeast cells was photographically monitored using a "cryomicroscope" which is capable of controlling in a programmable manner both the temperature and the time rate of change in temperature of the cell suspension being studied. Multiple nonlinear regression analysis together with a thermodynamic model of cell water transport during freezing was then used to statistically  deduce the subzero temperature dependence of the cell water permeability. The water permeability process for S. cerevisiae being cooled at subzero temperatures was found to be rate-limited by the passage of water through either the plasmalemma, the cell wall, or a combination of these two permeability barriers. The hydraulic water permeability coefficient for yeast at 20 degrees C is approximately 1--2 x 10(-13) cm3/dyne sec, if extrapolation from subzero temperatures to room temperature is permissible, while the apparent activation energy governing the permeability process at subzero temperatures is approximately 45--68 kJ/mol (11--16 kcal/mol).


Evaluation of enzyme linked immunosorbent assay in the immunodiagnosis of schistosomiasis.

Eighty-six bilharzial patients divided into 5 clinical groups were studied. Enzyme linked immunosorbent assay (ELISA), indirect fluorescent antibody (IFA) and indirect haemagglutination (IHA) test were performed for all the patients. ELISA gave the most sensitive results (82.6% and 80.2% positivity rate in Egypt and Lille respectively), followed by IFA (79.1% positivity rate) and IHA (77.9% and 75.6% positivity rate in Egypt and Lille respectively). The humoral antibodies detected by all methods (ELISA, IFA and IHA) showed increasing values with the progress of the disease which is parallel to the antigenicity of the disease. Both the positivity rate and the mean value of antibody titres recorded by the 3 diagnostic techniques (ELISA, IFA and IHA) were significantly higher in mansoni than haematobium infection. This may be explained by species specificity of antibody response. The superiority of ELISA over IFA and IHA techniques was discussed.


Initiation of DNA synthesis on the isolated strands of bacteriophage f1 replicative-form DNA.

Viral and complementary strand circular DNA molecules were isolated from intracellular bacteriophage f1 replicative-form DNA. Soluble protein extracts of Escherichia coli were used to examine the initiation of DNA synthesis on these DNA templates. The initiation of DNA synthesis on f1 viral strand DNA was catalyzed by E. coli DNA-dependent RNA polymerase, as was initiation of f1 viral strand DNA isolated from mature phage particles. The site of initiation was the same as that used in vivo. In contrast, no de novo initiation of DNA synthesis was detected on f1 complementary strand DNA. Control experiments demonstrated that the E. coli dnaB, dnaC, and dnaG initiation proteins were active under the conditions employed. The results suggest that the viral strand of the f1 replicative-form DNA molecule carries the same DNA synthesis initiation site as the viral strand packaged in mature phage, whereas the complementary strand of the replicative-form DNA molecule carries no site for de novo primer synthesis. These in vitro observations are consistent with the simple rolling circle model for f1 DNA replication in vivo proposed by Horiuchi and Zinder.


Q beta-defective particles produced in a streptomycin-resistant Escherichia coli mutant.

This paper describes Q beta noninfectious particles produced at 41 degrees C in a streptomycin-resistant Escherichia coli mutant which is temperature sensitive for suppression of a nonsense codon. The noninfectious particles resembled Q beta under the electron microscope and contained coat protein molecules in an amount similar to the amount in Q beta. However, they did not adsorb to F-piliated bacteria, and they were deficient in both minor capsid proteins of Q beta, maturation (IIa) and read-through (IIb). Proteins IIa and IIb were not produced in Qbeta-infected mutant cells at 41 degrees C. In addition, instead of the 30S RNA of Q beta, a shorter RNA, which sedimented mainly at 23 S, was found in the defective particles. The results are discussed in relation to the roles of proteins IIa and IIb of Q beta.


Development of Escherichia coli virus T1: escape from host restriction.

The bacterial virus T1 grows interchangeably on different Escherichia coli strains (C, B, and K). This implies that T1 has an efficient mechanism to overcome the host restriction barrier. The DNA of T1 was found to be methylated independently of the hosts. The percentage of N6-methyladenine varied from 1.6 to 1.8, and the 5-methylcytosine content varied from 0.1 to 0.4%. In contrast, the range in percentage of N6-methyladenine and 5-methylcytosine found in the hosts was 0.7 to 2.4 and 0.0 to 1.1, respectively.


Bacteriophage T4 alt gene maps between genes 30 and 54.

Two- and three-factor crosses proved that the T4 alt gene, which codes for a virion-associated NAD+:protein ADP-ribosyltransferase, is located between genes 30 and 54.


Replication of bacteriophage phiK duplex replicative-form DNA in dnaB and dnaC mutants of Escherichia coli.

We have directly tested the effects of host cell DNA synthesis mutations on bacteriophage phiK replicative-form (RF) DNA replication in vivo. We observed that phiK RF DNA replication continued at normal rates in both dnaB and dnaC mutant hosts under conditions in which the activities of the dnaB and dnaC gene products were shown to be markedly reduced. This suggests that these two host proteins are not essential for normal phiK RF DNA replication. In control experiments we observed markedly reduced rates of phiK RF DNA replication in temperature-sensitive dnaG and dnaE host mutants, indicating that the products of these genes are essential. Thus, the mechanism of DNA chain initiation in vivo on the duplex RF DNA templates of isometric phages such as phiK apparently is different from that on the similar templates of isometric phages such as phiX174. The implications of this difference are discussed in the text.


DNA synthesis in Escherichia coli cells infected with gene H mutants of bacteriophage phi X174.

Escherichia coli cells infected with gene H mutants of bacteriophage phi X174 produce two types of particles. The 110S particles contain single-stranded circular DNA; the 110S particles are not infectious, although their DNA is infectious for E. coli spheroplasts. The second type of particles, 70S particles, contain a fragment of single-stranded DNA ranging from 0.2 to 0.5 genome in length. This fragment DNA anneals only to restriction enzyme fragments of replicative-form DNA from the portion of the molecule corresponding to the origin and early region of phi X174 single-stranded synthesis, although full-round single-stranded DNA synthesis is occurring in the H mutant-infected cells. Different H mutant phages produce different proportions of 70S to 110S particles; those mutants producing the most 70S also exhibit the largest amount of degradation of intracellularly labeled DNA during infection. These results suggest that in H mutant-infected cells, full-length single-stranded DNA is synthesized; varying amounts of degradation of the single-stranded material occur, and the resulting fragment DNA is subsequently incorporated into 70S particles.


Vertical transmission of group B Streptococcus. Relation to intrauterine fetal monitoring.

A prospective study of 70 mother-infant pairs was designed to evaluate vertical transmission of group B Streptococcus (GBS) in relation to the use of intrauterine fetal monitors (IUFMs). Multiple-site cultures obtained from mothers during the intrapartum interval and those obtained from infants on day 1 and at discharge or day 4 were plated on a selective medium. Thirteen (27.1%) of 48 IUFM-exposed women vs seven (31.8%) of 22 non-IUFM-exposed women had GBS at one or more sites. The GBS colonization with maternally concordant serotypes occurred in eight (61.5%) of 13 infants born to GBS-colonized, IUFM-exposed women vs two (28.6%) of seven infants born to GBS-colonized, non-IUFM-exposed women. While this suggests that vertical transmission of GBS is enhanced by IUFM placement, the differences in these infant rates were not statistically significant.


Oral amphotericin for candidiasis in patients with hematologic neoplasms. An autopsy study.

Autopsy examinations were conducted in 72 patients with hematologic malignant neoplasms who received antibacterial therapy before their deaths. These patients were participants in a large double-blind study designed to assess the efficacy of oral amphotericin B in decreasing the incidence of candidal infection. The patients received either 50 mg of amphotericin B orally four times a day, or they received a matching placebo. Eight of 33 patients (24%) who had received placebo and two of 39 (5%) who had received amphotericin had histopathologic evidence of disseminated candidiasis. We conclude that in these patients with hematologic malignant neoplasms who received antibiotics within two weeks of death, the concomitant oral administration of amphotericin was effective in decreasing the incidence of systemic candidal infections, indicating that the gastrointestinal tract serves as a portal of entry for Candida albicans.


Hepatitis B surface antigenemia in renal transplant recipients. Increased mortality risk.

To determine effects of circulating hepatitis B surface antigen (HBsAg) on patient survival following renal transplantation, we studied 168 recipients of cadaveric grafts in whom HBsAg status was defined at transplantation by comparison of survivorships determined by actuarial life-table methods. Survival in HBsAg-positive recipients, as compared with those who were HBsAg-negative at the time of transplantation, was markedly diminished. Although graft survival appeared to be favored in the HBsAg-positive group in the early posttransplant period, the apparent protective effect was lost at nine months after grafting and thereafter, due to increased mortality in the HBsAg-positive recipients. Deaths in the antigenemic subjects were principally due to infections other than hepatitis and to cardiovascular events. We conclude that preexisting HBs antigenemia forebodes an ominous outcome for immunosuppressed renal transplant recipients, although hepatic disorders do not account for most deaths.


Primary cutaneous nocardiosis.

Seven patients with primary cutaneous nocardiosis were seen during a 20-month period. Six patients had a history of local trauma to an extremity or did frequent yard work. The spectrum of clinical disease included cellulitis, pustules,pyoderma, or lymphocutaneous syndrome mimicking sporotrichosis. Evidence of Nocardia infection elsewhere was absent, and no patient had a serious underlying disease. Nocardia brasiliensis was isolated from four cases, and N asteroides was identified in the other three patients. In two patients, disease resolved without chemotherapy, while a short course of a sulfonamide with or without trimethoprim therapy was successful in the remaining cases. The relatively benign nature of this disease, the usual failure to obtain cultures of superficial skin infections, and our accumulation of seven cases in less than two years suggest this disease is more common than previously recognized.


Emergency treatment of asthma. A comparison of two treatment regimens.

The effectiveness of epinephrine was compared to that of a combination of epinephrine and aminophylline in the initial treatment of acute asthma. Forty-four patients with 51 episodes of acute asthma were evaluated. Peak flow spirometry served as an objective measure of airway resistance, and theophylline levels were determined at fixed intervals throughout the study. Epinephrine and aminophylline were not found to be superior to epinephrine alone. There was no correlation between mean serum theophylline levels and the magnitude of improvement. Rapidity of emergency department discharge and frequency of admission was independent of treatment method. The failure of epinephrine-aminophylline to effect more rapid or profound improvement in pulmonary function might suggest that epinephrine alone, or an equivalent sympathomimetic is a rational choice in the initial treatment of acute asthma.


Immunohistologic findings in the glomerular crescents in various renal diseases.

Renal tissues from 37 patients with glomerulopathies involving glomerular crescents were investigated using an immunofluorescence technique. Immunohistologic findings revealed two kinds of crescents, those with fibrinogen deposits (active), and those without (inactive). The degree of IgG deposition in glomeruli with active crescents was much higher than in glomeruli with inactive crescents in acute glomerulonephritis (AGN) and chronic glomerulonephritis (CGN). Active crescents were observed only in biopsy specimens taken within three months after the onset of acute glomerulonephritis or the acute exacerbation of chronic glomerulonephritis. These findings suggest that in AGN and CGN active crescents occur in an earlier stage of glomerular lesions and a more active stage in the immunological process than inactive crescents. The significance of active crescents in SLE, diabetic nephropathy and nephropathy associated with rheumatic arthritis was not evaluated due to the small number of patients.


Evaluation of 99mTc-albumin dilution curve with simultaneous injection of 99mTc- and 131I-albumins.

A critical comparison of radiocardiograms with 99mTc- and 131I-albumins was made to determine whether or not 99mTc-albumin can be used as a tracer for measuring blood volume (BV) and cardiac output (CO). Simultaneous injection of both isotopes was given to twenty-two patients to obtain radiocardiograms for each by means of a single scintillation counter with two pulse height analyzers. Such method allows for the determination of the energy levels for both 99mTc and 131I. The radiocardiographic pattern was somewhat different between 99mTc- and 131I-radiocardiograms: The ratio of left to right peak height was 16% on the average lower in the 99mTc-radiocardiogram in the 131I-one. The reason for the lower ratio is explained by greater absorption of 99mTc-radioactivity within the soft tissue interposing between the scintillation probe and heart. The values for 99mtc-derived blood volume were 7.4% on the average larger than those for 131I-derived blood volume (p less than 0.01), although the correlation was excellent (r = 0.98, p less than 0.01). There was no systemic difference between cardiac output determined by 99mTc- and 131I-albumins since the overstimation of blood volume can compensate for the understimation of CO/BV by 99mTc-study. In view of its rapid extravasation, 99mTc-albumin is not so appropriate tracer for accurate measurements of BV and CO/BV, although it may be used for measurement of cardiac output.


Impedance cardiography for the assessment of cardiac function during exercise.

1. Impedance cardiography proved valuable to estimate the stroke volume and other cardiac parameters noninvasively, when the electrical resistivity of the blood was corrected with hematocrit. 2. Cardiac response to graded exercise on a bicycle ergometer up to the maximal load in healthy young and active middle-aged subjects was divided into four phases from the changes in cardiac parameters such as stroke volume, cardiac output, ejection index and height of the atrial wave. 3. The ischemic patients did not tolerate well the exercise after the second phase or drift phase of exercise. 4. Physical training enhanced the cardiac function in work through increase in cardiac output and work load.


Post-operative long-term adjuvant immunochemotherapy with mitomycin-C, PSK and FT-207 in gastric cancer patients.

Long-term adjuvant immunochemotherapy carried out on the gastrectomized patients with stomach cancer was reported. The protocol comprises the administration of large-dose of Mitomycin-C (20+10) mg just after gastrectomy and the long-term administration of PSK, FT-207 or (PSK+FT-207). Almost no side effects were observed. According to the studies on the immunological parameters, the increased reactions in PPD skin test and lymphocyte blastogenesis induced by PHA and PWM were observed remarkably in group (P+F) 3 or 6 months after gastrectomy. As for the survival rate, group (P+F) showed the most preferable results at one year in stage IV, at two years in stage III and at three years in all the stages, respectively after gastrectomy.


Organ and tumor specificity of colon mucoprotein antigen in a rat model.

We purified a high-molecular-weight colon mucoprotein antigen (CMA) from normal F344 rat colon and from a transplantable dimethylhydrazine-induced colon carcinoma. Chemical analysis of the mucins showed similar amino acid and carbohydrate compositions. This finding was in contrast to the major differences that occurred in the composition of human CMA as a consequence of neoplasia. Immunohistochemical techniques were used in an examination of organ and tumor specificities. A New Zealand White rabbit antiserum against purified normal rat CMA, appropriately absorbed, detected a normal colon-specific determinant(s). The organ-specific determinant was lost as a consequence of neoplasia. However, a new tumor-specific determinant was then detected. CMA may have a potential role as an organ-specific marker of the neoplastic process.


Growth of normal and malignant human mammary epithelial cells in culture.

Normal and malignant human mammary epithelial cells were placed in culture. Normal cells were recovered from late-lactation milk and breast fluids, and malignant cells were isolated from primary breast tumors by collagenase digestion. The concentration of cells obtained from breast fluid samples was inversely proportional to the volume of fluid secreted. Most of these cells adhered rapidly to the substrate, did not replicate, displayed Fc receptor-dependent phagocytic activity, and were thus identified as macrophages. The remaining cells grew out into large islands comprised of one or two distinct morphologic types of mammary epithelial cells. Optimum growth of these cells was obtained in medium buffered to pH 6.8, and the epidermal growth factor markedly prolonged the exponential growth phase of the cells. Two morphologically distinct populations of epithelial cells were also observed in cultures established from individual breast tumors. Growth of the malignant cells was relatively unaffected by the pH of the culture medium, and the cells were unresponsive to exogenously added hormones. Overgrowth of malignant epithelial cells in primary cultures by stromal fibroblasts was retarded by replacement of standard growth medium with fresh medium containing a serum substitute; growth of the malignant epithelial cells was unaffected. A feeder layer of mitomycin C-treated human fibroblasts increased the plating efficiency of both normal and malignant cells in primary culture and also facilitated passage of these cells to secondary and tertiary cultures.


Bone tissue: composition and function.

A relatively small amount of bone tissue is present in the human body in view of the tissue's structural and chemical importance. The disparity between the constant appearance of mineralized bone matrix, regardless of source, under the electron microscope and the variability of previously reported analyses of bone specimens of a much larger size was disturbing and led to studies of the content and distribution of water in bone tissue and in whole bone specimens. Water in premineralized bone matrix is largely replaced by the non-crystalline (amorphous) and crystalline mineral phase without changes in the overall volume of bone matrix or that of whole bones, nor in the volume and position of collagen fibrils. This resistance to volume and fibril position change during mineralization is probably unique to bone tissue, and permitted critical water analyses of whole bone specimens of varying porosity. When marrow and bone tissue weight and volume analyses were compared with the specific gravity of whole bone specimens, constancy of the composition of bone tissue in these specimens was demonstrated. This result led to a concentration of our studies on the cells that synthesize and control this unique tissue. Currently, the investigations of this laboratory are oriented to study of the enzyme chemistry of isolated bone cells and of whole bone specimens, as well as the localization of matrix-bound and membrane-bound enzymes in bone tissue by histochemical techniques suitable for electron microscopy.


[Recent aspects on the use of specific antisera in diagnosis and therapy of acute lymphatic leukaemia in infants (author's transl)].

Acute lymphatic leukaemia (ALL) in infants is a very heterogeneous disease from the immunological point of view. Phaenotyping of leukaemic cells in 138 patients via different immunmarkers has so far allowed subdivision into 6 subgroups. Clinical risk factors and correlations with the immunologically defined subgroups, as well as their significance for prognosis, are demonstrated with the help of 61 examined ALL patients. 90-95% of all acute lymphatic leukaemias can presently be identified via heterologous antisera. After corresponding absorption, these antisera show high cytotoxicity against ALL cells without displaying any aggressivity towards normal haemopoetic stem cells. The authors make therapeutic use of anti-T cell and anti-common ALL (cALL) globulin in autologous bone marrow transplantation. This method involves withdrawal of bone marrow from patients with ALL in complete remission, the marrow being then fractionated, treated with antiserum to destroy residual leukaemic cells, and then cryopreserved. The leukaemia-free bone marrow will thus be available for transplantation in relapse.


[Monitoring of remission quality in acute leukemia of childhood by repeated determinations of B-cell-allo-antigen on lymphoblasts (author's transl)].

B-Cell-allo-antigen was demonstrated on lymphoblasts of children with acute lymphoblastic leukemia and on 14 permanent growing lymphoblastoid cell-lines. Pooled sera of 4 selected multiparas and a rabbit antiserum were used as specific antisera. Antigen was more effectively detected with heterologous antibodies than with human antisera. Primary lymphoblasts were characterized by B-cell-allo-antigen and lack of membrane immunoglobulins and Fc-receptors on the same cell. The quality of remission, i.e. the early diagnosis of relapse was better surveyed by immunological characterization than by cytological assessment. The bone marrow was examined in 8 patients with initial leukemia, in 13 patients with partial remission and in 30 patients in complete hematological remission. A striking discrepancy between immunological and cytological examination was observed in the bone marrow 4 weeks after inital anti-leukemic chemotherapy. At that time an increased number of blast-typed cells could still be counted in the immunological evaluation, although the cytological evaluation revealed already a complete hematological remission.


Experimental rabies in skunks: immunofluorescence light and electron microscopic studies.

Striped skunks (Mephitis mephitis) were inoculated into the abductor digiti quinti muscle with street rabies virus isolated from salivary glands of rabid skunks. Using the immunofluorescence technique, antigen was detected in muscle cells at the inoculation site before it was detected in the central nervous system. Neurons and their processes in nearly all regions of the brain, spinal cord, cerebrospinal ganglia, and peripheral nerves contained antigen in terminal stages of the disease. Electron microscopically, matrix (viral nucleocapsid), virions, and anomalous viral products were mainly in neuronal perikarya and dendrites, and less often in myelinated axons. Matrices, virions, and crystalloid structures were in muscle fibers at the inoculation site. Viral budding occurred on endoplasmic reticulum, neurotubules, and neuronal plasma membrane. In the brain and dorsal horn of the spinal cord, virus budded from the postsynaptic and adjacent dendritic or perikaryal plasma membrane. There was simultaneous esotropic uptake of these particles by adjacent axon terminals. The results strongly suggest that direct transneuronal transfer of virus from perikarya and dendrites to adjacent axon terminals is a mechanism in dissemination of rabies in the central nervous system of striped skunks. Variation in the length of the incubation period may be due partly to replication or virus in myocytes at the inoculation site and subsequent transfer to peripheral nerves.


Mesangial glomerulonephropathy with deposition of IgG, IgM, and C3 induced by mercuric chloride: a new model.

A mesangial glomerulonephropathy, characterized by the deposition of rat IgG, IgM, and C3 in the glomerular mesangium, was produced in Wistar rats by a prolonged administration of mercuric chloride (HgCl2). The HgCl2 was dissolved in sterile distilled water (0.2 mg. per ml.), and a group of 15 male Wistar rats was given injections subcutaneously three times a week on alternate days at a dosage of 0.15 mg. per 100 gm. of body weight for 27 weeks. A control group of nine rats was given injections of distilled water only. Mesangial glomerulonephropathy developed in 12 of 15 rats injected with HgCl2 and was characterized by the following: (1) coarse granular and nodular deposition of rat IgG, IgM, and C3 in the mesangium of all glomeruli, (2) absence of staining for rat albumin, IgA, and fibrin, (3) presence of electron-dense deposits in the mesangium, (4) focal and segmental proliferation of the mesangial matrix, (5) interstitial inflammation, (6) tubular atrophy, and (7) deposition of periodic acid-Schiff-positive material in the medulla adjacent to the thin limbs of the loops of Henle. Glycosuria and a slight increase in proteinuria were observed transiently in some rats. The blood urea nitrogen levels were normal in all rats. Eluates from the kidneys with heavy mesangial deposits contained rat IgG. However, the eluted antibody failed to react with normal rat kidney tissue components. None of the above findings were present in the control rats. The study provides a model of a mesangial nephropathy that seems to be immunologically induced; however, the mechanism for the formation and deposition of the immune deposits containing rat IgG, IgM, and C3, and the nature of the antigen(s) have not been elucidated.