Immunoglobulins in the jejunal mucosa in adult coeliac disease and dermatitis herpetiformis after the reintroduction of dietary gluten. Cells containing immunoglobulin (IgA, IgG, IgM) have been measured and the distribution of extracellular and epithelial cell immunoglobulin assessed in treated patients with adult coeliac disease (ACD) and dermatitis herpetiformis (DH) before and after gluten was reintroduced to the diet. Patients with ACD and DH frequently had IgM and IgG cells above the normal range even before re-exposure to gluten, although the range of IgA cells was normal. In both diseases IgA and IgM cells increased after gluten with a proportionally greater rise in the latter, so that numbers of IgM cells, but not of IgA, exceeded the control range in all but one patient. There were increased quantities of IgA and IgM extracellularly in the lamina propria and in epithelial cells after challenge with gluten. Third component of complement was also found in some biopsies after re-exposure to gluten. These findings support the suggestion that gluten induces a humoral immunological response within the small intestinal mucosa and that both IgA and IgM systems are involved.


Brain monoamines in seizure mechanism (review). A majority of experimental studies have strongly suggested that catecholamine and/or 5-HT play an important role for regulating the seizure susceptibility. As mentioned previously, however, the relative significance of individual monoamine has not yet been fully clarified. Since it is well known that the interaction between catecholaminergic and serotonergic neuronal activity is quite complex, a causal relationship between each monoamine and seizure susceptibility cannot be easily established if results are obtained from the whole brain study on pharmacologic manipulation of experimental seizures. The extensive study, including the monoamine turnover rates and concentrations in specific brain regions, may help to delineate such a correlation in the future. Further, when an object of study is concerned in excitability of the central nervous system, it should be necessary to elucidate the mutual relationship of monoamines to the other putative neurotransmitters (e.g., acetylcholine, amino acid). Does there exist an abnormal metabolism of monoamines in the brain of epileptic patients? If so, how is it related to the elaboration or maintenance of epileptic seizures? Unfortunately, we have no sufficeint information on the monoamine metabolism of epileptic patients.


[Kidney transplantation from a nephrological-urological viewpoint--results and problems. 2. Diagnosis and therapy after transplantation, complications, long-term results]. Indications, selection of donor and recipient, medical and surgical management and complications, problems of organ procurement. Renal transplantation has become routine therapy. Organs are predominantly obtained from cadavers, transplantations from living donors are rarely indicated. Advances in preservation methods have improved organ quality and prolonged storage time. Selection of the most suitable recipient is based on histocompatibility matching. Blood transfusions before transplantation seem to improve the results. Recognition of a rejection crisis is primarily based on clinical symptoms. Persistent rejection calls for prompt explantation and the patient has to return to dialysis. Infections, serum-hepatitis and gastro-intestinal bleeding are the most common complications. Late complicatons are diabetes mellitus, cirrhosis of the liver, osteopathy, recurring glomerulonephritis, and, rarely, malignomas. Transplantation frequency in the Federal Republic of Germany could be increased by more awareness of physicians and a better knowledge of the general public about the need for cadaver donors.


Kinetics of early insulin response by the isolated and perfused rat pancreas. Suggestion for two different actions of glucose on the insulin releasing process. The permissive action of glucose (or glyceraldelhyde) is necessary to the glucagon induced insulin release. By collecting every 15 seconds the venous effluent of the perfused and isolated rat pancreas it was observed that the insulin response to glucagon (2 microgram/ml) was immediate if the pancreas was preperfused with low concentration of glucose (5 mM) or glyceraldelhyde (2,5 mM). On the other hand glucagon alone elicited no response, and the insulin discharge occurred 60 to 90 seconds after the addition of glucose (5 mM) or glyceraldelhyde (2,5 mM) this time being probably allowed to the metabolism of the sugar. The pancreatic response to 15 mM glucose occurred also 60 to 90 seconds after the stimulus. On the other hand when the medium contained a low concentration of glucose (or glyceraldelhyde) increasing the glucose concentration by 10 mM provoked an immediate insulin release. This suggests that glucose has two actions differing by their lag phase. One, "permissive", apparent after some delay, mimicked by glyceraldelhyde, necessary for glucagon induced insulin release, is mediated probably by the metabolic products of the sugar. The second, "triggering", initiates instantaneously the insulin release but appears dependent on the first action.


Endocrine cells in oxyntic mucosa of a dog 5 years after pancreatectomy. Immunofluorescence shows that the oxyntic mucosa of a dog depancreatized for 5 years and having a poorly-controlled diabetes has more glucagon- and somatostatin-containing cells than the mucosa of a control dog. At the ultrastructural level, 4 endocrine cell types are identified: A-, A-like, D- and enterochromaffin-like (ECL) cells, with increased numbers of A-, A-like and D-cells in gastric glands of the depancreatized dog, together with a higher concentration of immunoreactive glucagon in the gastric mucosa. The increase in A-, A-like and D-cells is compatible with: a) a change induced by the diabetic state itself; b) a hyperplasia secondary to the loss of corresponding pancreatic cells. At any rate, the fact that A-, A-like and D-cells increase parallely may indicate that these three cell types are functionally related one with another.


Hypersensitivity of the human gonadotrophs to the repeated administration of small doses of LH-RH. The functional capacity of the gonadotrophs was assessed by repeated stimulation with small doses of LH-RH (5 microgram intravenously at 2-hour intervals for 3 injections) in normal women during the early and late follicular phases of the menstrual cycle. The results were compared to those obtained when a single dose (100 microgram) of the neurohormone was administered. During the early follicular phase, the release of LH and FSH remained about equal after the 3 successive injections of the small and after the large dose of LH-RH. During the late follicular phase, the release of LH and fsh increased progressively after the repeated administration of the 5 microgram of the neurohormone while the large dose induced a more pronounced and a more sustained pituitary response. This hypersensitivity of the gonadotrophs is observed when the E2 concentrations are higher than in the early follicular phase of the menstrual cycle.


Formation of a RNA polymerase sub-assembly composed of subunit alpha from Escherichia coli and of subunit beta from Micrococcus luteus. Functionally equivalent subunits of RNA polymerase from Micrococcus luteus and Escherichia coli differ from each other in many molecular and antigenic properties. In spite of these differences, subunit alpha from E. coli and subunit beta from M. luteus form a complex alpha2beta, when incubated together. This complex binds rifampicin tightly, which the isolated subunits do not. The hybrid complex is very similar in its properties to the complex alpha2beta formed only from E. coli or M. luteus subunits. Since the sub-assembly alpha2beta from E. coli is reported to be an obligatory intermediate in the assembly process of complete RNA polymerase, the newly described hybrid sub-assembly may function similarly as an intermediate in the formation of the hybrid form of RNA polymerase described earlier.


[DNCB contact sensitization: theoretical and clinical aspects (author's transl)]. In clinical immunology the overall assessment of cellular immunocompetence has become a subject of increasing interest. Beside the development of various sophisticated in vitro methods, simple skin tests remain a basic instrument. Contact sensitization to dinitrochlorobenzene (DNCB) has proved to be a reliable parameter in detecting impaired cel-mediated immunity, in that previous exposure to this antigen is rare and the epidermal application induces a primary immune response in the overwhelming majority of normal individuals. The morphology of the skin reactions corresponds to that of contact eczema (allergic contact dermatitis). A review of the literature shows that various techniques of the test are applied which complicate the interpretation and comparison of different results. The aim of this study was to report our own experiences with DNCB contact sensitization in normal individuals and in patients with Crohn's disease and complex glomerulonephritis and to discuss practical problems in test application; in particular the differentiation between toxic and allergic (delayed hypersensitivity) dermatitis, the choice of the sensitizing and rechallenge dose, the principles of recording successful sensitization, the influence of the inflammatory response upon sensitization, side effects, and precautions. Finally a modified test system is recommended.


Regulation of IgG antibody synthesis by a B cell component. An immunoglobulin-G recruiting component (GRC) prepared from splenic B cells of antigen-primed mice was shown to be effective in recruiting more IgG plaque forming cells than normally appear among splenic cells experiencing a primary immune response. GRC caused increases in all classes of IgG PFC except perhaps IgG3, and the largest improvements were in IgG1 and IgG2a. GRC is synthesized by IgG2a-bearing cells and is effective at 96-120 h after spleen cells have been exposed to antigen. It is incapable of substituting for allogeneic effect factor, and the latter apparently must have its input on antibody producing cells before GRC can act. Together the data suggest that during a primary immune response a definite number of splenic B cells become poised for synthesizing IgG antibody, but only a portion of them are able to secrete. Apparently, the quiescent cells among them can be activated to secrete by exposure to GRC, A B cell product.


Partial characterization of a T cell-derived factor that suppresses the initiation of the humoral immune response in vitro. Specific anamnestic stimulation of spleen cells from mice immunized 7 days earlier with horse erythrocytes (HRBC) generated the release of a soluble factor that was capable of suppressing the initiation of the in vitro primary gammaM immune response to sheep red blood cells (SRBC), as well as to the immunogen that elicited its formation. Moreover, the suppressive macromolecule (mol. wt yields to 34,000), derived from antigen-activated, HRBC-primed T lymphocytes (but not B cells), inhibited the secondary gammaM and gammaG anti-SRBC plaque-forming cell responses of SRBC-primed spleen cells. The active material was resistant to treatment with DNase and RNase, but was inactivated by protease (10 microgram/ml, 30 min) or exposure to mild heat (56 degrees, 30 min). The antibody initiation suppressor factor (AISF) was concentrated and partially purified by gel filtration, followed by poly-acrylamide gel electrophoresis.


Thymic-dependent anti-hapten response in congenitally athymic (nude) mice immunized with DNP-thymosin. Immunization of congenitally athymic (nu/nu) and adult thymectomized, irradiated bone marrow, reconstituted (TxBm) mice with DNP5-thymosin (dinitrophenylated-bovine thymosin fraction 5) was found to elcit IgM and IgG anti-DNP plaque-forming cells in these animals. Further studies indicated that this response was antigen specific and not due to polyclonal activation. Since the hormonal properties of the thymosin were retained following linkage with hapten and DNP-thymosin was immunogenic in CBA/N and CBA/N female X DBA/2 male)F1 male mice, animals previously shown to have an X-linked inability to respond to thymus-independent antigens, it was concluded that DNP-thymosin functions both as a hormone and as a T-dependent antigen in eliciting an immune response in nu/nu and TxBm mice. Additional support for this conclusion was provided by the demonstration that DNP-thymosin could specifically prime for and elicit an anamnestic response in nu/nu mice. These results indicate that further investigation of the immune activities of DNP-thymosin may provide valuable insight in characterizing the maturation of helper T cells and their subsequent interaction with B cells.


Suppression of cell-mediated immune responses by dextran sulphate. The effect of high molecular weight dextran sulphate (DS) on cell-mediated immune responses was studied. Two criteria were used to assess cell-mediated responses, skin graft rejection and intra-dermal (i.d.) skin tests. Mice receiving allografts and DS displayed a significant increase in graft survival time compared with untreated control animals. Similar results were obtained when the grafted animals differed at only the H-Y locus. Guinea-pigs sensitized with Mycobacterium tuberculosis and treated with DS showed a greatly reduced delayed skin test response when challenged with purified protein derivative. DS was capable of suppressing this response even though, by the same route, dosage, and time of administration, it was previously shown to enhance a cell-mediated response in guinea-pigs sensitized with a different antigen. The results of this study indicate that the adjuvant DS is capable of suppressing cell-mediated immune responses in mice and guinea-pigs.


Dislocation of the calcaneum.

Dislocation of the calcaneum. Dislocation of the tarsal bones is uncommon; isolated dislocation of the calcaneum has been reported only four times. This paper presents a case of dislocated calcaneum and reviews the literature.


Migraine as a sequela of blunt head injury. Evidence that a blunt head injury can engender a migrainous state in susceptible individuals is provided by a series of 13 patients who developed recurring attacks of classic migraine after suffering such injury. Among a group of patients with post-traumatic headache of more than one year's standing, the preponderant type of headache was found to be of the migrainous variety. The significance of these findings is discussed.


Demonstration of enterobacterial common antigen by bacterial agglutination. Potent antisera against the enterobacterial common antigen (ECA) agglutinate R bacteria of the Enterobacteriaceae family that possess unimpaired R-core structures of the Escherichia coli R1 or E. coli R4 core type. In these strains, known to be ECA immunogenic, ECA is most probably linked to the lipopolysaccharide R core. R mutants of other core types (e.g., Salmonella Ra, E. coli R2 or R3) or R mutants with incomplete core structures of the E. coli R1 type, as well as an rfaL mutant deficient in the O-translocase system, agglutinate to a much lesser extent or not at all. All the later mutants are nonimmunogenic; they possess the ECA in a free form, not linked to the R core. None of the S forms tested from many different enterobacterial genera was found to be agglutinable with the ECA antiserum. The dynamics of the ECA agglutinin formation in rabbits parallels the ECA hemagglutinin formation, indicating that the same antibody class might be involved in bacterial agglutination and hemagglutination.


Chemotactic factor produced by Candida albicans. Candida albicans was found to produce a substance that was chemotactically active for guinea pig polymorphonuclear neutrophils. The chemotactic factor was detected in culture filtrates of organisms grown under aeration and incubated at 37 degrees C for at least 12 h. Nutrients found to be essential for the production of chemotactic factor included glucose, yeast extract, and a mixture of amino acids. Several strains of C. albicans isolated from humans were tested, and varying degrees of chemotactic activity were found to be associated with the culture filtrates. Only one of the eight isolates did not produce a measurable amount of chemotactic activity. Culture filtrates remained chemotactically active after several cycles of freezing and thawing and after heating at 90 degrees C for 10 min. Substantial evidence is presented that the chemotactic activity is not dependent upon activation of complement.


Role of phagocytosis in mouse virulence of Salmonella typhimurium recombinants with O antigen 6,7 or 4,12. The quality of lipopolysaccharide has previously been shown to influence the mouse virulence of Salmonella so that strains with O antigen 4,12 were more virulent than their O-9,12 sister strains. Immunosuppression did not alter this O-antigen-dependent difference in virulence. I have now constructed smooth O-4,12 and O-6,7 sister hybrid strains of Salmonella typhimurium. No other phenotypic differences were found between these strains; they were all "common antigen" positive. In intraperitoneal infection, the O-4,12 strains were more mouse virulent than their O-6,7 sisters. The difference in virulence correlated with a difference in clearance rates; the O-6,7 hybrids were removed from the blood more rapidly than their O-4,12 sisters. No natural bactericidal antibodies were found in the sera of the mice.


Selective in vitro response of thymus-derived lymphocytes from Treponema pallidum-infected rabbits. The blastogenic response of nylon wool-separated peripheral-blood lymphocytes from Treponema pallidum-infected rabbits was tested in vitro with mitogens and T. pallidum antigens. The mitogenic response of the enriched T-cell population to concanavalin A and phytohemagglutinin was depressed during the first 3 to 4 weeks of infection, similar to the pattern observed with unfractionated cells. Shortly thereafter, levels of blastogenesis returned to values of uninfected cultures. Enhanced blast transformation was seen immediately when purified T-cells from infected rabbits were exposed in vitro to T. pallidum antigens. Although these relatively high levels of blastogenesis were maintained for the duration of the experiment, cultures of unfractionated lymphocytes from infected rabbits did not exhibit an increased blastogenic response to the same antigen preparation until 3 to 4 weeks after infection. Autologous serum from infected rabbits decreased the lymphocyte response to T. pallidum antigen. The stimulatory effects of anti-immunoglobulin G and lipopolysaccharide on nylon wool-fractionated or unfractionated lymphocytes from both infected and control rabbits were similar throughout the course of infection. During the first 6 weeks of experimental disease, there was a 25 to 31% increase in the number of lymphocytes circulating in the peripheral blood of T. pallidum-infected rabbits.


Autoantibodies detected in rabbits hyperimmunized with group A, C, and G streptococcal vaccines. Rabbits hyperimmunized with group A, C, and G streptococcal vaccines developed autoantibodies with affinities for different tissues (smooth muscle, cytoplasmic, and myocardial antibodies) and for autologous proteins (anti-albumin and anti-immunoglobulin antibodies). The presence of anti-albumin and smooth muscle antibodies, associated with a high level of immunoglobulin, suggests the development of hepatic disorders in the hyperimmunized rabbits.


Induction of cell-mediated immunity to Mycobacterium lepraemurium in susceptible mice. A mouse strain (CB6) that is highly susceptible to Mycobacterium lepraemurium was infected with 10(8) bacilli into the hind footpad. These mice developed cell-mediated immunity to M. lepraemurium, as expressed by the development of a granulomatous lesion at the site of inoculation in normal but not in T-lymphocyte-depleted mice, a proliferative response in the paracortical zone of the draining lymph node, delayed-type hypersensitivity to a sonic extract of M. lepraemurium, and immunopotentiation of the delayed hypersensitivity response to sheep erythrocytes. Resistance to a second challenge infection with M. lepraemurium was not demonstrated.


Group B streptococcal long-chain reaction. The streptococcal long-chain reaction was adapted for the measurement of type-specific antibodies to group B beta-hemolytic streptococci (GBBHS). Rabbit antisera incubated with homologous but not heterologous GBBHS produced chains that were 18 to 33 times longer than chains produced by normal rabbit sera. The long chains were easily apparent, in most instances, by scanning the slides. Human sera with mouse protective and opsonic activity against GBBHS serotype Ia produced chains that were always significantly longer than those produced by incubation in nonimmune human sera. Absorption of rabbit or human sera with homologous but not heterologous organisms inhibited the capacity to induce the formation of long chains. The long-chain assay is a simple, rapid, and reproducible test that could constitute a valuable tool for the rapid identification of anti-GBBHS antibodies.


Immune response to Cryptococcus neoformans soluble polysaccharide: immunological unresponsiveness. Mice injected with 100 to 800 microgram of Cryptococcus neoformans soluble polysaccharide showed a reduced ability to produce antibody after a challenge immunization with polysaccharide emulsified in Freund incomplete adjuvant. These animals were considered immunologically unresponsive. Animals given an initial injection of 25 or 50 microgram of polysaccharide responded to a challenge immunization in the same manner as control animals. Reversion of unresponsive mice to antibody production without further antigenic stimulation did not occur during a 12-week experimental period. These animals exhibited a partial response to challenge immunization 8 weeks after induction of unresponsiveness, and they were fully responsive to challenge immunization at 12 weeks. Animals given a single dose of 0.1, 0.4, or 1.6 microgram of polysaccharide produced a marked anamnestic response after challenge immunization. Repeated injections of subimmunogenic doses of polysaccharide did not produce a marked anamnestic response and would induce unresponsiveness only when the cumulative dose reached 100 to 400 microgram of polysaccharide, suggesting that injected cryptococcal polysaccharide might be sequestered in some manner until an amount of antigen sufficient for induction of unresponsiveness is accumulated. This possibility was confirmed by immunofluorescence studies that revealed a long-term deposition of polysaccharide in the tubular epithelial cells of the kidney.


Vero response to a cytotoxin of Escherichia coli. A cytotoxin was found in culture filtrates of a number of Escherichia coli strains that differed from the known heat-stable and heat-labile enterotoxins of E. coli. It was cytotoxic for Vero but not for Y-1 or CHO cells, and its effect on Vero was distinctly different from that of heat-labile enterotoxin. It was labile to heat and antigenically different from heat-labile enterotoxin, and membrane filtration indicated a molecular weight of 10,000 to 30,000.


Antibody response in rabbits to immunization with Mycobacterium leprae. Mycobacterium leprae purified from liver tissue of an infected armadillo (the A/10 preparation) was tested for antigenic composition by immunization of rabbits and characterization of the antibody response by crossed immunoelectrophoresis. The rabbit antisera detected seven distinct components in the M. leprae preparation. This number is far lower than in similar experiments with other mycobacteria. The M. leprae sonic extract gave far fewer lines after polyacrylamide gel electrophoresis and staining with Coomassie brillant blue than sonic extracts prepared from BCG, M. smegmatis, and M. phlei adjusted to the same protein concentration based on the Folin assay. The seven components detected in M. leprae cross-reacted extensively with M. avium, BCG, M. lepraemurium, M. smegmatis, and Nocardia asteroides. The seven components are involved in immune reactions in leprosy; antibodies against all of them were demonstrated in sera from patients with lepromatous leprosy, but the specificity of the antibodies varied from patient to patient. The reason for the demonstration of so few antigenic components and some of the implications of these findings for the use of armadillo-grown M. leprae to develop specific skin test reagents and in other aspects of leprosy research are discussed.


Anabolic potential of virulent Treponema pallidum. Acrylamide gel autoradiography of 3H-labeled proteins from Treponema pallidum demonstrates that virulent treponemes incubated in vitro synthesize a spectrum of high-molecular-weight proteins. A comparison of the protein profiles of T. pallidum with the Reiter treponeme shows that T. pallidum possesses significant anabolic competence.


Failure of endotoxin to protect C3H/HeJ mice against lethal x-irradiation. C3H/HeJ mice are more sensitive to lethal X-irradiation and cannot be protected by pretreatment with endotoxin, compared to the C3HeB/FeJ strain. Lack of a granulopoietic response in C3H/HeJ mice is evident.


Guanine-plus-cytosine composition of five group B streptococcal serotypes. The percent guanine-plus-cytosine content of deoxyribonucleic acid of each of the five serotypes of group B streptococci was determined by thermal denaturation. The range of guanine-plus-cytosine content was 35.1 to 36.9%, with a mean value of 35.9%. These values suggest a genetic homogeneity to the serotypes of the group B streptococci.


Chemical adhesion in dentistry.

Chemical adhesion in dentistry. The terms used in the chemistry of adhesion are reviewed as well as the role played by surface energies, viscosity, surface roughness, etc. The similarity of chemical bonds in substrate and adhesive which favours adhesion in general, is very rare in dentistry. It is shown how and when a chemical bridge between two different types of materials works in dentistry. Finally, it is made evident why binding to dental hard tissues will not be permanent.


Adhesion to teeth.

Adhesion to teeth. Using standard surface chemical techniques it can be demonstrated that under physiological conditions, the adhesive properties of human enamel and dentine are created by the volumetrically small organic portion of these tissues. As far as their adhesiveness is concerned undamaged teeth therefore do not differ considerably from other types of biological tissues. Generally speaking the presence of low energy, organic films will act to prevent formation of strong adhesive contacts between tooth surfaces and e.g. bacteria, food stuffs or restorative materials. By certain types of direct surface treatments the adhesiveness of tooth surfaces can be changed temporarily or permanently in ways that will either increase or decrease the chances to obtain strong adhesive contacts. Under physiological conditions teeth are covered with rapidly forming salivary films which reduces the possibility to obtain strong adhesive contacts with teeth. If clinically useful adhesion to tooth surfaces is to be obtained the teeth involved must therefore be kept isolated from saliva during the operative procedures.


The aetiology of dental caries--an outline of current thought. Nearly 90 years after being put forward by W.D. Miller, the acidogenic theory of dental caries aetiology is supported by a wealth of experimental evidence. However, while explaining the basic mechanism of caries, it does not indicate how the effect of the mechanism is modified to give the observable pattern of caries attack. This explanation is attempted on the basis of the interaction of two groups of variable factors. Firstly, those which affect the tooth's resistance to caries attack, which include the chemical, microstructural and morphological nature of the enamel surface. Secondly the factors which determine the cariogenicity of the tooth's environment. These include the nature of the diet, plaque and saliva. Finally the caries initiation is considered at an atomic level in terms of the dynamic ionic exchange between enamel and plaque. This brief review highlights the need for considerably more research to shed more light on the nature of the variable aetiological factors of caries.


Role of T and adherent cells in in vitro response of nude mouse spleen cells to bacterial lipopolysaccharide. Thy 1.2-positive cells and adherent cells were depleted from nude mouse spleen cell suspensions by treatment with anti-Thy 1.2 antisera plus complement and by passage through Sephadex G-10 column, respectively. The results of 3H-TdR uptake and appearance of IgM containing cells induced in vitro by lipopolysaccharide (LPS) were similar in both untreated and depleted spleen cells. Our findings confirm T and adherent cell independence of LPS response in nude mice.


Metabolically dependent and independent cell surface antigens. In contrast to metabolically dependent tissue-specific cell surface antigens (MDA) which are available for reactions with antibodies only on surfaces of metabolically active cells, the availability of the universally distributed blood group A or Forssman-type antigens on cell surfaces was found to be independent of the metabolic activity of cultured cells. In case of the MDA, cytotoxic reactions were induced by antibodies alone, resulting in a disorganization of the cellular sheet without a significant release of radioactive label. Radioactive release in this cytotoxic reaction was increased by the addition of complement without additionally affecting the degree of cell sheet disorganization. In case of Forssman and blood group A antigens, such morphologically demonstrable cytotoxicity required complement, and the resulting pathology was always accompanied by extensive release of cellular contents. The ability of anti-MDA antibodies to induce cytotoxic reactions in the absence of complement may be related to a vital role of MDA in cellular function.


Adjuvant properties of Micropolyspora faeni. The adjuvant properties of Micropolyspora faeni, an important source of antigenic material in the production of farmer's lung, were evaluated by comparing antibody- and cell-mediated immune responses of rabbits to bovine serum albumin (BSA) incorporated in complete Freund's adjuvant (CFA), incomplete Freund's adjuvant (IFA) and incomplete Freund's adjuvant with 5-10 mg/ml homogenized M. faeni (MFA). Rabbits immunized with BSA in CFA or MFA developed significantly increased antigen-induced macrophage migration inhibition, lymphocyte stimulation, and delayed skin reactivity when compared to those immunized with BSA in IFA. No similar adjuvant effect on specific antibody production was observed in rabbits immunized using BSA in MFA. These data suggest that M. faeni can act as a selective immunologic adjuvant for delayed hypersensitivity. This adjuvant property might be important in the induction of mononuclear cell infiltrates seen in human hypersensitivity pneumonitis.


Silent renal involvement in systemic lupus erythematosus. 20 patients with active SLE without clinical evidence of renal involvement underwent percutaneous renal biopsy. 12 had varying proliferative changes on light microscopy. Of 19 ultrastructural examinations performed only 3 had no electron-dense deposits. Serum C3 and C4 levels were 63 +/- 8 and 8 +/- 2 mg% in patients with subendothelial deposits, compared to 142 +/- 27 and 27 +/- 6 mg%, respectively, in patients without deposits  (p less than 0.01). All patients with diffuse proliferative changes had subendothelial deposits; however, one with normal light microscopy and another with focal proliferation also had them. It is concluded that no variant of lupus nephropathy can be excluded on clinical grounds alone.


Host response to treponema pallidum infection. II. Rabbit leukocyte migration inhibition in the presence of homologous organ extracts. Peripheral leukocytes of 67 rabbits infected intratesticularly with Treponema pallidum, Nichols' strain for various lengths of time were examined by the migration inhibition test for their response to extracts of normal rabbit heart, skin, brain and T. pallidum antigen. A control group of 14 animals injected intratesticularly with extract of normal rabbit tests was similarly examined for the leukocyte response to the same antigens except the brain extract. The percentage of infected animals responding to T. pallidum antigen with significant migration inhibition varied from 13 to 31. Transitional cellular response to heart and skin but not brain was observed (12-28%). Leukocytes of all but 2 control rabbits responded to the organ extracts within the limit of 2 SD. The response of the infected animals to the homologous organ extracts may suggest that during infection, lymphocytes are activated by the host tissue antigens.


The course of Hymenolepis nana infections in thymus-deficient mice. The kinetics of infection with Hymenolepis nana was examined in normal thymus-deficient mice. Following inoculation with 5 cyticercoids, the number of adult lumen-dwelling H. nana in congenitally thymus-deficient (nude) mice ultimately was at least 75 times the maximum infection intensity of normal thymus-bearing mice or thymus-reconstituted nude mice. Similar results were obtained following inoculation of H. nana eggs into nude mice; although thymus-bearing mice eliminated their infections, nude mice harbored more than 1,000 adult worms throughout the 7 weeks of the experiments. These data show that in mice, immunity is not generated against H. nana in the absence of thymus function. The data also confirm and establish the requirement of cysticercoid development in the intestinal mucosa for stimulation of a protective immune response against H. nana.


Proteolysis of bovine immunoglobulins.

Proteolysis of bovine immunoglobulins. Bovine serum IgG1, colostral IgG1 and serum IgG2 with anti-ferritin activity were digested with pepsin or trypsin. Their fragments were characterized by immunoelectrophoresis, gel electrophoresis and gel filtration; their ferritin-binding ability was determined. The kinetics of proteolysis were established by measuring the appearance of free amino groups. No differences were observed between serum and colostrum IgG1. IgG1 was more susceptible to pepsin, and IgG2 to trypsin. This became evident from both the amount of intact IgG determined by gel electrophoresis, immunoelectrophoresis or gel filtration, and from the kinetics of the appearance of amino groups. A model is presented to explain the size, mobilities and properties of the obtained fragments.


Anatomical distribution of T and B lymphocytes identified by immunohistochemistry in the chicken spleen. Antisera against chicken bursal and thymic lymphocytes were raised in rabbits. They were made specific for the target T or B cells by appropriat absorption and then used for immunohistochemical identification of T and B lymphocytes in tissue sections of chicken spleen, applying the unlabeled antibody enzyme method. Lymphocyte reacting with anti-chicken-thymus globulin predominated in the periarteriolar lymphatic sheaths and in the red-pulp cords. A few of these lymphocytes were localized between the periellipsoid lymphatic tissue and in germinal centers. The periellipsoid lymphocytes around the Schweigger-Seidel sheaths consisted of lymphocytes positive with anti-chicken-bursa-cell globulin. The majority of lymphocytes in the germinal centers were labeled also by anti-chicken bursa-cell globulin. Age-dependent changes in the anatomical localization of T and B lymphocytes are described between late embryonic development and an age of 2 years. The different conditions of avian and mammalian T and B cell distribution in the spleen were considered.


Some observations on fuzzy diagnosis and medical computing. Wechsler has advocated the use of fuzzy logic for medical diagnosis, but it is pointed out that simpler systems have proved more successful than is apparent from his paper. The proposal raises a broad range of issues concerning the role of sophisticated information-processing techniques in the development of medical computing. These issues are discussed and it is suggested that a more evolutionary view of this development would be of greater benefit in the long run.


Potential of the amino acid homology studies: homologies found between beta-galactosidase and lac repressor of E. coli. Significant amino acid homologies were found between beta-galactosidase fragments and lac repressor of E. coli using a sliding match according to Greller and Erhan (1974). Since both of these proteins can recognise and bind galactose moiety, we propose that the homologous regions represetn galactose binding site(s) on both proteins. Possible application of homology studies to problems of protein and nucleic acid chemistry is also discussed.


Circulating immune complexes in patients with malignant lymphomas and solid tumors. Sera from 50 patients with Hodgkin's disease, 78 patients with non-Hodgkin non-leukemic malignant lymphomas, and 75 patients with different types of solid malignant tumors were investigated for the presence of immune complexes using the (125I) C1q-binding test. All patients were untreated. An increased serum C1q-binding activity was found in 22% of the patients with Hodgkin's disease, 35.9% of the non-Hodgkin lymphoma patients and in 37.3% of the patients with solid tumors. The C1q-binding material detected in the patients' sera had properties similar to those of immune complexes. On sucrose density gradient it sedimented as a 10-30 s material. It contained IgG which were dissociated under acid conditions. Passage through anti-IgG immunoabsorbent removed its C1q-binding properties. A prevalent association was found between the presence of serum immune complexes and disseminated disease stages in all the patient groups included. A similar association was found between the presence of serum immune complexes and general symptoms among the malignant lymphoma patients. The nature of the antigens involved in the complexes remains unknown.


Secondary prevention of drug dependence through the transcendental meditation program in metropolitan Philadelphia. Using mailed questionnaires, the Transcendental Meditation program was investigated as an approach to the secondary prevention of substance dependence. Significant differences were found between a random sample of meditators and matched control subjects in usage levels and percentage of users for almost all legal and illegal drugs. Meditators had used more prescribed psychoactive medications before learning meditation, but usage levels returned to normal soon after starting. Former users of alcohol, cigarettes, or illegal drugs achieved remarkable abstinence records. For most substances the amount of decrease was positively correlated with degree of participation in the program and the length of time meditating.


Methadone maintenance and narcotic blocking drugs. Appendix. The research history of norcodeine as a narcotic blocking drug is presented. Because of the limitations found for norcodeine usage, naloxone, cyclazocine, and naltrexone have recently received attention, and the results of research with them are detailed.


The structure of urban mortality. A methodological study of Hannover, Germany. Part I. An analysis is made of the association between density factors and the profile of cause specific death rates in small areas (Bezirke) of Hannover, Germany, in 1968-69. Evaluation of urban ecological methods and logic is done to determine the plausibility of making meaningful statements about cause specific death risks due to areal attributes. Efforts are made to develop an appropriate multiple indicator structural equation model to determine if underlying patterns of mortality can be discerned. Evidence is found that measures of crowding, apart from classical density, predict a general mortality component.


The epidemiology of diseases of the hip: a review of the literature. Diseases of the hip are significant public health problems. Fractures and osteoarthrosis of the hip are common in older age-groups, while congenital dislocation of the hip, Legg-Calvé-Perthes disease, and slipped epiphysis affect a certain proportion of infants, children, and adolescents. The distribution of these five common hip diseases in the population is reviewed, aetiological hypotheses are discussed, and gaps in knowledge identified. Particular attention is focused on the roles of mechanical stress and hormones, since there is evidence for the involvement of one or both of these factors in the aetiology of each of the diseases. Many characteristics of their descriptive epidemiology have still to be explained, however, and it is clear that more research is needed.


The influence of membrane lipid composition and procaine on hyperthermic death of cells. The mechanism of hyperthermic killing, a component of some cancer therapy, is not known. Cell-survival curves during hyperthermic exposure can be used to elucidate the effects of membrane modifying procedures on cell death. Experiments were designed to test whether procedures that were reported to increase membrane fluidity also resulted in increased killing on hyperthermic exposure. An E. coli K12 mutant, K1060, is used to predictably alter the degree and amount of unsaturated fatty acids incorporated into membranes. Changing from an 18:1 to an 18:3 unsaturated fatty acid increases killing. Decreasing the amount of unsaturated fatty acid cells incorporated by increasing growth temperature decreases killing. Procaine, a drug known to decrease membrane viscosity, increases heat killing. These data are most simply explained by the hypothesis that membrane disorganization occurs as a result of temperature increasing to a point where a lipid transition causes a membrane structural change, which results in cell-death.


Pituitary-gonadal feedback in testicular disorders. Stimulation response of the pituitary to LHRH and testicular biopsy were investigated in 17 infertile men in comparison to 10 healthy controls. A significant elevation of basal FSH and LHRH-stimulated FSH serum levels was observed in Sertoli cell only syndrome and spermatogenetic arrest, while LH and testosterone remained normal. The LHRH test seems to be of value for differentiation of pretesticular, testicular and posttesticular infertility.


Description and interpretation of structural changes in canine kidneys after preservation with a hyperosmolar solution and reperfusion. The effects of hypothermic preservation of canine kidneys for 24 hours by the use of the Sacks solution, and additional autogenous reperfusion were studied by means of submicroscopic morphological methods. While preservation with the Sacks solution was to ensure normal organization of the lipoprotein structures of the basement and cell membranes, in case of additional reperfusion, even of short duration, a substantial or complete loss of anisotropy of the cytomembranes was demonstrable. The present report is the first to be published in the literature on polarization microscopic studies concerned with the effects of kidney preservation and of reperfusion.


Transmethylation hypothesis of schizophrenia: methionine and nicotinic acid. The transmethylation hypothesis of schizophrenia was reviewed with considerations that large doses of methionine when combined with a monoamine oxidase inhibitor lead to exacerbation of psychotic symptoms in a significant percentage of chronic schizophrenic patients. It was noted that nicotinic acid in the dosage of 3,000 mg/day can neither prevent nor counteract the psychopathology thus induced.


Results of immediate reconstruction following surgery for advanced cases of cancer of the cheek, jaw and lip. Results of immediate reconstruction of the defects resulting from excision of cancer of the cheek, lower jaw, upper jaw and lip in 13 patients using temporal island, deltopectoral and cervicoacromial flaps have been described. The surgical techniques have been outlined. Previous radiotherapy is not a contraindication for primary excisional and reconstructive procedures although the incidence of postoperative gangrene of the flap was greater in these cases.


Bladder response to klebsiella infection: a scanning electron microscopy study. The early response of rat bladder epithelium to Klebsiella pneumoniae was examined using scanning electron microscopy. Increased bacterial adherence with time after challenge paralleled the occurrence of strand formation and ulceration on the bladder epithelium. The early bladder response to the bacteria consisted of altered epithelial membrane structure, strand formation, and exfoliation of some epithelial cells. These observations indicate that K. pneumoniae causes alterations in the bladder epithelium which may trigger an intrinsic bladder defense mechanism consisting of strand entrapment of bacteria, epithelial cell exfoliation, and elimination of attached bacteria via voiding.


Modification of Trichoplusia ni nuclear polyhedrosis virus passaged in vivo. A designated strain of plaque-purified Trichoplusia ni nuclear polyhedrosis virus (NPV) was used to initiate a serial passage series in vivo. The virulence of polyhedra inclusion bodies (PIB) and the distribution of MP (many PIB) and FP (few PIB) strains in hemolymph were monitored. When virus was passaged per os as PIB, there was no major change in virulence after 15 passes in insects. Plaque analysis of hemolymph from the infected insects indicated that there was no selection for the FP strain of virus. There was little overall change in the infectivity of nonoccluded virus (NOV) after 16 passes by injection into T. ni larvae; however, plaque analysis of the hemolymph from the infected insects indicated progressive selection of FP virus. It is postulated that when PIB are the vehicles of virus transmission, there is no progressive accumulation of noninfectious FP PIB; however, when NOV are the vehicles of virus transmission, FP NOV production displaces MP virus synthesis. This may result from a growth advantage of the FP NOV over the MP NOV as shown in vitro and from the noninfectious nature of FP PIB.


On the mechanism of the persistence of lymphocytic choriomeningitis virus in the continuous cell line Detroit-6. A persistent lymphocytic choriomeningitis virus, noninfectious for mice, was revealed in the continuous human cell line, Detroit-6. The virus was detected in the cell monolayer by an indirect immunofluorescence test and in the cell homogenate by a complement-fixation test. Between 30 and 80% of the cells produced viral antigen in subsequent passages of the culture. Thymidine analogues (BrDU and IDU) stimulated the synthesis of antigen. DNA of the persistently infected cell line was transfected to the mouse lymphoid cell line, L1210, which became persistently infected with the virus.


Effect of diazepam on uterine activity during labor. The effect of diazepam on uterine activity was assessed in a series of 30 patients in established spontaneous labor. The action of diazepam on the frequency and amplitude of contractions, uterine tone and fetal heart rate was monitored internally. An i.v. dose of 10 mg diazepam significantly decreased uterine activity by its direct effect on contraction frequency. No effect on uterine tone was seen. This effect of diazepam on contraction frequency was additional to its tranquilizing and muscle relaxant action.


Mesenteric lymphadenitis due to Yersinia pseudotuberculosis. A case of mesenteric lymphadenitis caused by Yersinia pseudotuberculosis is described in a young adult. The diagnosis was suggested by the histological appearance of the lymph nodes, and confirmed by serological tests. This is the first report of mesenteric lymphadenitis due to Y. pseudotuberculosis in Israel.


Humps on young human and rabbit articular cartilage. Scanning electron microscopic studies revealed numerous humps on the surface or articular cartilage from a 7 years old boy and from 1 month old rabbits. This shows that the surface topography of articular cartilage in young individuals is different from that of older individuals (human and rabbit) where the surface is beset by numerous pits. In 2 months old rabbits an intermediate situation was witnessed, for both shallow pits and occasional humps were present. Ridges and undulations were not seen on the articular surface of the 7 years old boy, except near the cut edge of the specimen. They were, however, found on air-dried specimens of 1 month old rabbit cartilage, but were absent from specimens dried by the critical-point method. The collective evidence supports the idea that ridges and undulations are not a constant feature of the normal articular surface, but that such features are artefactual, a typical or pathological.


Nocardicin A, a new monocyclic beta-lactam antibiotic V. In vivo evaluation. Nocardicin A is a new monocyclic beta-lactam antibiotic which provides a potent therapeutic effect in mice experimentally infected with gram-negative bacilli. When given subcutaneously to mice, the therapeutic effect of the drug was stronger than had been anticipated from in vitro studies. Nocardicin A was more potent in therapeutic effect than carbenicillin against infections due to Pseudomonas aeruginosa, Proteus mirabilis, Pr. vulgaris, Pr. rettgeri and Pr. inconstans, and was similar in effect to carbenicillin against infections due to Escherichia coli in mice. In addition, nocardicin A proved to be active against infections due to Serratia marcescens and other organisms resistant to beta-lactam antibiotics. When nocardicin A was given subcutaneously to mice, blood and hepatic levels of the drug were higher than those of carbenicillin.


A colI-specified product, synthesized in newly infected recipients, limits the amount of DNA transferred during conjugation of Escherichia coli K-12. The amount of ColI DNA transferred between mating cells of Escherichia coli K-12 increased about fourfold when rifampin-resistant donors were mated with sensitive recipients in the presence of the drug. Conjugational synthesis of ColI in dnaB recipients, shown primarily to reflect conversion of the transferred DNA into double-stranded material, was also enhanced when the recipients were treated with either rifampin or streptomycin. It is suggested that the amount of ColI transfer is normally limited by the synthesis of one or more proteins in the newly infected recipients. The protein is thought to be plasmid-specified because rifampin also quadrupled transfer to UV-irradiated recipients which were deficient in the transcription of the resident DNA. Successive strands of ColI appear to be transferred discontinuously, because the transferred DNA accumulated in normal and rifampin-treated recipients in the form of circular and linear monomeric units. Although rifampin treatment of recipients also increased transfer of a second Ialpha plasmid, R144drd-3, by about four times, the drug failed to cause a substantial increase of Flac transfer in comparable matings.


Genetics of nitrofurazone resistance in Escherichia coli. Wild-type Escherichia coli cells are sensitive to nitrofurazone (NF) and many other nitrofuran derivatives. A variety of evidence indicated that these compounds are converted to toxic "active" metabolites by reductases present in the bacteria. Sensitive E. coli K-12 acquired threefold-greater resistance to NF in one mutational step. These partially resistant mutants could undergo a second mutation that made them 10 times as resistant as the wild type. Mutation of wild-type strain K-12 to the higher level of resistance in a single step was not observed. The first mutational step was associated with partial loss of reduced nicotinamide adenine dinucleotide phosphate-linked, O(2)-insensitive NF reductase activity, and the second step was associated with loss of the remaining activity. The two-step mutants did, however, contain other NF reductases that were inhibited by O(2) and reduced NF only under anaerobic conditions. We designated the genes that control reductase activity "nitrofuran sensitivity genes" (nfsA and nfsB). Thus, wild-type strains are nfsA(+)nfsB(+), and the resistant double mutants are nfsA nfsB. A variety of crosses established that these genes are both located close to gal, that the most probable sequence is lac nfsB gal nfsA, and that the single-step mutants with an intermediate level of resistance are nfsA nfsB(+). The nfsA(+)nfsB strains contained about 70 to 80% of the wild-type reductase I activity-apparently enough to confer wild-type sensitivity. This reductase activity was resistant to 2 M urea. The nfsA nfsB(+) strains had only 20 to 30% of the wild-type activity, and this residual activity was sensitive to 2 M urea.


Genetics of Treponema: relationship between Treponema pallidum and five cultivable treponemes. Three genetically distinct groups of treponemes have been identified by saturation reassociation assays using 125I-labeled treponemal DNAs. The three groups are (i) virulent Treponema pallidum (Nichols strain), (ii) T. phagedenis and its biotypes Reiter and Kazan 5, and (iii) T. refringens biotypes Nichols and Noguchi. There is no detectable DNA sequence homology (less than 5%) among the three groups. The groups have distinct guanine + cytosine contents: 52.4 to 53.7% for T. pallidum, 41.5% for T. refringens, and 38 to 39% for T. phagedenis.


Bacteriophage conversion of heat-labile enterotoxin in Escherichia coli. A temperate phage designated obeta1 (omicron beta) was mitomycin C induced and isolated from heat-labile enterotoxin (LT)-producing Escherichia coli E2631-C2. Phage obeta1 infected the nonlysogenic, nontoxigenic, mitomycin C-sensitive strain of E. coli K-12 (CSH38) and converted it to lysogeny and enterotoxigenicity. After the establishment of lysogeny, E. coli CSH38(obeta1) produced produced LT and phage particles at maximal levels following mitomycin C induction. The LT Tox+ character is carried by the temperate phage obeta1.


Regulatory properties of araC(c) mutants in the L-arabinose operon of escherichia coliB/r. Merodiploids containing a high-constitutive and a low-constitutive araC(c) allele were assayed for constitutive expression of the ara operon. Low-constitutive araC(c) alleles either were unable to repress the constitutive rate of ara operon expression exhibited by by high-constitutive araC(c) alleles or achieved a partial repression of the high-constitutive rate of operon expression. Either mutation to a low-constitutive araC(c) mutant resulted in a partial or complete loss of repressor function, or subunit mixing between the two araC(c) mutant proteins resulted in a partial or complete dominance of the high-constitutive araC(c) allele. Five of the six araC(c) alleles tested allowed a partial induction of the ara operon in cya crp background. In general, a higher level of ara operon induction was achieved in the cya crp background by high araC(c) alleles than by low araC(c) alleles. Furthermore, several araC(c) mutants exhibited decreased sensitivity to catabolite repression, particularly in the presence of inducer. The results suggest a model in which certain araC(c) gene products can achieve ara operon induction in the presence of either arabinose (inducer) or catabolite activator protein-cyclic adenosine monophosphate, whereas the wild-type araC gene product requires the presence of both of these factors for operon expression.


Low-temperature conditional cell division mutants of Escherichia coli. Fifteen low-temperature conditional division mutants of Escherichia coli K-12 was isolated. They grew normally at 39 degrees C but formed filaments at 30 degrees C. All exhibited a coordinated burst of cell division when the filaments were shifted to the permissive temperature (39 degrees C). None of the various agents that stimulate cell division in other mutant systems (salt, sucrose, ethanol, and chloramphenicol) was very effective in restoring colony-forming ability at 25 degrees C or in stimulating cell division in broth. One of these mutants, strain JS10, was found to have an altered cell envelope as evidenced by increased sensitivity to deoxycholate and antibiotics, as well as leakage of ribonulcease I, a periplasmic enzyme. This mutant had normal rates of DNA synthesis, RNA synthesis, and phospholipid synthesis at both the nonpermissive and permissive temperatures. However, strain JS10 required new protein synthesis in the apparent absence of new RNA synthesis for division of filaments at the permissive temperature. The division of lesion in strain JS10 is cotransducible with malA, aroB, and glpD and maps within min 72 to 75 on the E. coli chromosome.


Uptake of extracellular biotin by Escherichia coli biotin prototrophs. Uptake of exogenous biotin by two Escherichia coli biotin prototroph strains, K-12 and Crookes, appeared to involve incorporation at a fixed number of binding sites located at the cell membrane. Incorporation was characterized as a binding process specific for biotin, not requiring energy, and stimulated by acidic pH. Constant saturation quantities of exogenous biotin were incorporated by these cells, and the amounts, which were titrated, depended on whether the cells were resting or dividing. Resting cells incorporated exogenous biotin amounting to 2% of their total intracellular biotin content. Fifty percent of the exogenous biotin was incorporated into their free biotin fraction, and 50% was incorporated into their bound biotin fraction. On the other hand, dividing cells incorporated exogenous biotin into all of their intracellular sites, 88% going into the intracellular-bound biotin fraction, and 12% going into the free biotin fraction. Calculations suggested that each cell contained approximately 3,000 binding sites for biotin. It was postulated that biotin incorporation sites might have been components of acetyl coenzyme A carboxylase located at or near the membrane.


Arrangement of protein I in Escherichia coli outer membrane: cross-linking study. The arrangement of protein I in the outer membrane of Escherichia coli was investigated by cross-linking whole cells, isolated cell wall, protein-peptidoglycan complexes, and protein I released from peptidoglycan with NaCl. Both cleavable azide cross-linkers and imidoester reagents were used. The data presented suggest that protein I exists in the outer membrane as a trimer.


Immunochemical analysis of inner and outer membranes of Escherichia coli by crossed immunoelectrophoresis. Isolated membrane fractions of Escherichia coli K-12 yielded complex immunoprecipitate patterns when Triton X-100 and sodium dodecyl sulfate extracts were examined by crossed immunoelectrophoresis with antienvelope immunoglobulins. Twelve of the 46 antigens in the immunoprecipitate patterns of inner (plasma) membranes were identified by zymograms and/or by the use of specific antisera. The following enzyme activities were detected in immunoprecipitates: 6-phosphogluconate dehydrogenase (EC 1.1.1.43); adenosine triphosphatase (EC 3.6.1.3); glutamate dehydrogenase (EC 1.4.1.4), two separate components; malate dehydrogenase (EC 1.1.1.37); dihydroorotate dehydrogenase (EC 1.3.3.1); succinate dehydrogenase (EC 1.3.99.1); lactate dehydrogeanse (EC 1.1.1.27); reduced nicotinamide adenine dinucleotide dehydrogenase (EC 1.6.99.3); protease (EC 3.4.21.1); and glycerol 3-phosphate dehydrogenase (EC 1.1.99.5). The corresponding immunoprecipitate pattern for isolated outer membranes consisted of at least 25 discrete antigens and differed strikingly from that obtained with inner membranes. Two major immunogens were identified as lipopolysaccharide and Braun lipoprotein. A protease-active immunoprecipitate was also detected in this fraction, but attempts to identify the Rosenbusch matrix protein in the crossed immunoelectrophoretic profile were unsuccessful.


Effect of thymine concentration on cell shape in Thy- Escherichia coli B/r. Cells of a thymineless mutant of Escherichia coli B/r are shown to change their shape when the concentration of thymine in the growth medium is reduced. Electron micrographs of whole cells and isolated sacculi were used to make quantitative measurements of the changes in cell length and width which occur as a result of such a change in thymine concentration. The results showed that there is an increase in cell volume, which is due to an increase in cell width accompanied by a decrease in cell length. These changes were compared with the predictions of models which assume that cell shape is influenced by the chromosome replication cycle.


Interaction between two major outer membrane proteins of Escherichia coli: the matrix protein and the lipoprotein. The affinity to the matrix protein, one of the major outer membrane proteins of Escherichia coli, for the peptidoglycan was examined of extracting the cell envelope complex at 55 degrees C and 2% sodium dodecyl sulfate containing different amounts of NaCl. It was found that the matrix protein was extracted from the peptidoglycan of a mutant strain (lpo) that lacks another major membrane protein, the lipoprotein, at a lower NaCl concentration than was the matrix protein of the wild-type cell (lpo+). When the envelope fraction of the wild-type strain was treated with trypsin, which is known to cleave the bound-form lipoprotein from the peptidoglycan, the affinity of the matrix protein for the peptidoglycan decreased to the same level as that of the affinity of the matrix protein for the peptidoglycan of the mutant strain. It was further shown that the free-form lipoprotein was also retained in the matrix protein-peptidoglycan complex, although the extent of retention of the free form of the lipoprotein was less than that of the matrix protein. These results indicate that both the free and the bound forms of the lipoprotein are closely associated with the matrix protein and that the bound form of the lipoprotein plays and important role in the association between the matrix protein and the peptidoglycan.


F plasmid genes involved in the production of recombination-stimulating factor, control of sensitivity to some injurious agents, and chromosome replication in Escherichia coli K-12 HfrC. Three related F'arg+ plasmids isolated by Guyer and Clark were used to analyze some properties of strain MG751, a recipient derivative of an HfrC mutant (MG7) carrying a previously described pleiotropic mutation in the integrated F plasmid. Strains MG7 and MG751 both failed to produce recombination-stimulating factor, were sensitive to monofunctional alkylating agents and UV irradiation, and were temperature sensitive for growth and DNA synthesis. It was shown that these phenotypes are controlled by F plasmids genes (designated rsf, prt, and rep) that can be separated by deletion mutations occurring on the F plasmid.


Cloning of Beneckea genes in Escherichia coli. Genes from Beneckea harveyi, a luminescent marine bacterium, were cloned in Escherichia coli. This was done by producing randomly sheared fragments of Beneckea DNA and inserting them into the EcoRI site of plasmid pMB9 by the adenine-thymine joining procedure. The hybrid plasmids were used to transform E. coli C600 SF8. Among the transformants selected for tetracycline resistance, one clone that appeared to complement a leucine tb mutation was identified. The transformants were screened for the presence of Beneckea 5S genes. Four of these clones were analyzed in detail by hybridization with 16S, 23S, and 4S Beneckea RNA. The observations suggest that the ribosomal genes in Beneckea are linked, but are present in a different order than those in E. coli.


Identification of the structural gene for the hook subunit protein of Escherichia coli flagella. Previous studies showed that the structural gene for the flagellar hook subunit protein (molecular weight 42,000) was one of a group of flagellar genes located on the Escherichia coli genome near pyrC. Several lines of evidence indicate that the flaK gene is the structural gene for the hook subunit protein. Fla+ strains that were insensitive to chi infection could be isolated as revertants of an FlaK- amber mutant strain but from no other Fla- strain. The hook subunit proteins isolated from such chi-sensitive revertants of the FlaK- strain were shown to be antigenically and electrophoretically different from the hook protein isolated from the wild-type strain. Thus, reversion of a mutation in the flaK gene resulted in alteration of the structure of the hook protein. Furthermore, in programming experiments with hybrid lambda containing flagellar genes, lambdafla with flaK genetic activity programmed the synthesis of a 42,000-molecular weight protein, whereas lambdafla without flaK genetic activity did not.


R plasmic Rtsl-mediated production of extracellular deoxyribonuclease in Escherichia coli. Escherichia coli strains harboring Rtsl were found to excrete extracellular deoxyribonuclease. The DNase activity was greater in cells with pTW2, a mutant from Rtsl.


Primary structure of the Escherichia coli serotype K42 capsular polysaccharide and its serological identity with the Klebsiella K63 polysaccharide. The Escherichia coli K42 capsular polysaccharide consists of leads to 3)-alpha-D-Galp-(1 leads to 3)-alpha-D-GalUAp-(1 leads to 3)-alpha-L-Fucp-(1 leads to repeating units. The E. coli K42 and Klebsiella K63 antigens are serologically identical.


Preferential inhibitory action of sodium cholate on an Escherichia coli strain carrying a plasmid in an integrated state. Sodium cholate was shown to be preferentially more active on Escherichia coli strains carrying an integrated plasmid, i.e., on Hfr strains, than on their parental strains with or without a plasmid in an autonomous state.


Formation of N-carbamyl putrescine from citrulline in Escherichia coli. Decarboxylation of citrulline by Escherichia coli enzymes was presented. The N-carbamyl putrescine produced showed the same properties as those of synthesized authentic samples in column chromatography, paper chromatography, and paper electrophoresis.


Interaction between the fumarate reductase system of Escherichia coli and the nitrogen fixation genes of Klebsiella pneumoniae. For phenotypic expression of nif+Kp genes in Escherichia coli K-12, the anaerobic electron transport system to fumarate must be functional. The role of the fumarate reduction system is to energize the membrane and thus provide the energy necessary for nitrogen fixation.


Relationship between polyamines and macromolecules in germinating yeast ascospores. The accumulation of spermidine and/or spermine was not necessary for normal macromolecule biosynthesis or germination and outgrowth of Saccharomyces cerevisiae spores.


Conjugal transfer system of plasmid RP4: analysis by transposon 7 insertion. We have begun an analysis in Escherichia coli of the conjugal transfer functions of the broad-host-range plasmid RP4. We have isolated 19 tra mutants of RP4, generated by insertion of transposon 7, and mapped their insertion sites by restriction endonuclease analysis. These sites fall into two separate regions on either side of the kanamycin resistance determinant. The transfer rates of the mutants range from 10% of that of RP4 to an undetectable level. Spot tests with the P-1 pilus-specific phages PRR1, Pf3, and PR4 and electron microscopic examination for pili have classified the mutants into several phenotypes consistent with their having normal, retracted, or no pili. Analysis of transient plasmid heterozygotes, created by P1 transduction, divided the tra mutants into a minimum of five complementation groups. Some of these groups contain more than one phenotypic class and may represent more than one gene because of the possible polar and deletion effects of Tn7 insertion.


ColE1 plasmid mutants affecting growth of an Escherichia coli recB recC sbcB mutant. Three mutant derivatives of the plasmid ColE1 were found to reduce the formation of plasmidless cells in a recB recC sbcB cell population. An active plasmid role in the plasmid-host interaction is suggested.


F- mating materials able to generate a mating signal in mating with HfrH dnaB(Ts) cells. The purified outer membrane from F- (W1-3) cells was shown to inhibit mating effectively, but the purified cytoplasmic (inner) membrane did not. These membranes, heat-treated minicells, and ultraviolet-irradiated minicells were examined for their ability to generate a mating signal at 43 degrees C in mating with HfrH dnaB(Ts) cells. The outer and inner membranes and heat-treated minicells all failed to stimulate incorporation of radioactive thymine; only ultraviolet-irradiated minicells retained the ability to generate a mating signal for the donor to initiate transfer replication.


Single-strand breakage in DNA of Escherichia coli exposed to Cd2+. When a growing culture of Escherichia coli was exposed to 3 X 10(-6) M Cd2+, 85 to 95% of the cells lost their ability to form colonies on agar plates. Loss of viability was accompanied by considerable single-strand breakage in the DNA, with no detectable increase in double-strand breaks. A direct correlation appeared to exist between the number of single-strand breaks and the concentrations of Cd2+ to which the cells were exposed. Exposure of DNA in vitro to a Cd2+ concentration of 3 X 10(-6) M or higher, followed by sedimentation in alkaline sucrose gradients, demonstrated no single-strand breaks. Cadmium-exposed cells recovered viability when incubated in Cd2+-free liquid medium containing 10 mM hydroxyurea. During the early period of recovery, there was a lag in the incorporation of labeled thymidine, but cellular DNA, at least in part, appeared to be repaired.


Gene dosage effects of the structural gene for a lipoprotein of the Escherichia coli outer membrane. The gene dosage effects of the structural gene (lpp) for the lipoprotein of the Escherichia coli outer membrane were examined. A novel F-prime factor containing the lpp gene was constructed. The amount of the free-form lipoprotein in the merodiploid strain carrying the F-prime factor was found to be about two times as great as that in the corresponding haploid strain. On the other hand, the amount of the bound-form lipoprotein, which is vovalently linked to the peptidoglycan, was not significantly different in the merodiploid strain as compared with the corresponding haploid strain. The present results suggest that the lpp gene is expressed constitutively in contrast to another major protein of the E. coli outer membrane, tolG protein (protein II, D. B. Datta et al., J. Bacteriol. 128:834-841, 1976). The F-prime factor isolated may include a portion of the E. coli chromosome (located between 33 and 36 min on the genetic map) that is not covered by any other F-prime factor.


Identification of the Escherichia coli cell division gene sep and organization of the cell division-cell envelope genes in the sep-mur-ftsA-envA cluster as determined with specialized transducing lambda bacteriophages. From a lysogen with lambda integrated in the leu operon, specialized transducing phages that carry the cell division, murein biosynthesis, and envelope permeability genes located about 0.5 min to the right of leu were isolated. These phages were used to identify the previously undiscovered cell division gene sep. A genetic map proves that sep is located in the sequence leuA sep murE murF murC ddl ftsA envA. A physical map of this region was prepared by heteroduplex analysis of the phage DNAs. Overlapping segments of host DNA extended rightward for as much as 26.4 kilobase pairs from the prophage insertion point (thought to be in leuA) to include all the genes through envA.


Origin of hydrogen atoms in the fatty acids synthesized with yeast fatty acid synthetase. The mechanism of hydrogen incorporation into fatty acids was investigated with an enzyme preparation from baker's yeast. Fatty acids synthesized from malonyl-CoA and acetyl-CoA in the presence of D2O or stereospecifically deuterium-labeled NADPH were isolated and analyzed by mass chromatography to examine the localization of deuterium atoms in the molecule. The following results were obtained: 1. Hydrogen atoms from water were found on the even-numbered methylene carbon atoms (2-hydrogen atoms per carbon atom). The second hydrogen atom was incorporated as the result of hydrogen exchange phenomenon between the methylene group of malonyl CoA and water. 2. HB hydrogen of NADPH was used for beta-ketoacyl reductase. 3. HB hydrogen of NADPH was also used for enoyl reductase. 4. Hydrogen atoms from HB position of NADPH were found on the odd-numbered methylene carbon atoms (2-hydrogen atoms per carbon atom).


Purification and properties of alpha-mannosidase from bakers' yeast. The yeast alpha-mannosidase [EC 3.2.1.24] was purified 1160-fold from the crude extract of the autolysate. The purified preparation was practically free from alpha-glucosidase, beta-glucosidase, alpha-galactosidase, beta-galactosidase, beta-mannosidase, and beta-N-acetylhexosaminidase activities. After the separation of yeast mannan during the purification procedures the enzyme became unstable but could be stored at 5 degrees C for three weeks with 50% loss of activity. The purified enzyme hydrolyzed both aryl and alkyl mannosides, but hydrolysis of yeast mannan proceeded slowly. Yeast mannan and Zn2+ increased the enzyme catalyzed hydrolysis of p-nitrophenyl mannoside, whereas NaN3, monoiodoacetate and methyl alpha-D-mannoside acted as inhibitors. The molecular weight was estimated to be 450,000 by gel filtration.


Involvement of 30S ribosomal protein S1 in poly(U)-directed polyphenylalanine synthesis. The effect of 30S ribosomal protein S1 on poly(U)-directed polyphenylalanine synthesis was studied using a highly purified cell-free system which was devoid of endogenous S1. The system consisted of homogeneous preparations of EF-Tu, EF-Ts, and EF-G, and 70S ribosomes from which protein S1 had been removed by poly(U)-cellulose column chromatography. It was found that protein S1 was indispensable for translation of poly(U) by an S1-depleted system at low concentrations of poly(U). On the other hand, at higher concentrations of poly(U), a considerable amount of polyphenylalanine was synthesized in the absence of added S1. The stimulatory effect of S1 was observed at all Mg2+ concentrations examined but was most pronounced at 10 mM Mg2+. Some physicochemical properties of the protein were also studied. It was demonstrated that the protein has an elongated shape with an axial ratio of approximately 8.5.


Changes in the conformation and stability of 5 S RNA upon the binding of ribosomal proteins. The binding of ribosomal proteins L25, L18, and L5 to 5 S RNA results in a conformational change and a destabilization of the 5 S RNA molecule. The changes observed in the near ultraviolet circular dichroism (CD) spectra and in the melting profiles indicate an increase in base stacking uith an accompanying increase in asymmetry of the bases and a decrease in the conformational stability of the 5 S RNA. These results are consistent with the interpretation that the binding of these proteins increases the stacking of specific single-stranded bases in 5 S RNA and aligns them in helical arrays, resulting in a conformation which facilitates base-pairing with nucleotide segment(s) of the ribosomal 23 S RNA or the transfer RNA (or both). The simple and precise difference CD method described here is potentially useful for studying subtle conformational changes of other nucleic acid-protein interactions.


Purification and properties of a third form of anthranilate-5-phosphoribosylpyrophosphate phosphoribosyltransferase from the Enterobacteriaceae. Anthranilate-5-phosphoribosylpyrophosphate phosphoribosyltransferase was purified from the bacterium Erwinia carotovora, a member of the Enterobacteriaceae. The enzyme was homogeneous according to the criteria of gel electrophoresis and NH2-terminal amino acid sequence analysis. The molecular weight of the enzyme as determined on a calibrated Sephadex G-200 column was 67,000 +/- 2,000. Sodium dodecyl sulfate-polyacrylamide gels gave a subunit molecular weight of 40,000 +/- 1,000, suggesting that the enzyme was a dimer. A comparison of the NH2-terminal sequence of the enzyme with the (previously determined) homologue from Serratia marcescens, a monomer with a molecular weight of 45,000, showed that the larger Serratia subunit came into register with amino acid 14 of the Erwinia subunit. The register for the length of the known overlap, 26 amino acids, was highly conserved.


Processivity of DNA exonucleases.

Processivity of DNA exonucleases. A homopolymer system has been developed to examine the digestion strategies of DNA exonucleases. Escherichia coli exonuclease I and lambda-exonuclease, are processive enzymes. However, T7 exonuclease, spleen exonuclease, E. coli exonuclease III, the 3' leads to 5'-exonuclease of T4 DNA polymerase, and both the 3' leads to 5' and the 5' leads to 3' activity of E. coli DNA polymerase I dissociate frequently from the substrate during the course of digestion. Regions of duplex DNA are a dissociation signal for exonuclease I.


Identification of bound pyruvate essential for the activity of phosphatidylserine decarboxylase of Escherichia coli. Phosphatidylserine decarboxylase, an intrinsic membrane protein of Escherichia coli, catalyzes the decarboxylation of phosphatidylserine, the final step in the biosynthesis of phosphatidylethanolamine, the principal membrane lipid of this organism. The purified enzyme lacks the absorption spectrum characteristic of pyridoxal-containing enzymes, and it has now been found to contain bound pyruvate, the carbonyl function of which is essential for catalytic activity. The decarboxylase is inactivated by treatment with a number of reagents that attack carbonyl groups, including sodium borohydride. Reduction with tritiated borohydride leads to the introduction of stably bound radioactivity, which, after acid hydrolysis, has been identified as tritiated lactate by several chromatographic procedures and by treatment with lactate dehydrogenase. The enzyme resists inactivation by cyanoborohydride in the absence of substrate, but is readily inactivated by this reagent in the presence of phosphatidylserine. Under the conditions of treatment of neutral pH, cyanoborohydride does not react with carbonyl residues at an appreciable rate, but reduces imino groups much more rapidly. This finding, together with demonstrated dependence of the enzyme upon the carbonyl residue of pyruvate for activity, strongly suggests that a Schiff base is formed by addition of the amino group of phosphatidylserine to the pyruvate residue of the enzyme as an essential step in the action of the decarboxylase.


Interaction between DNA and Escherichia coli protein omega. Formation of a complex between single-stranded DNA and omega protein. Escherichia coli omega protein is found to form a complex with single-stranded DNA. The complex is stable in buoyant CsCl or Cs2SO4 density gradients. Addition of Mg(II) to the concentrated salt solutions, however, leads to the dissociation of the complex, even in the presence of EDTA in molar excess over Mg(II). The dissociated omega retains its enzymatic activity; the DNA recovered from the dissociated complex is indistinguishable from the original DNA. Exposure of the complex to alkali results in the cleavage of the DNA. This cleavage generates a 3'-hydroxyl DNA terminus, and the omega protein is found linked to the 5'-terminus, presumably covalently. Pronase digestion of the complex results initially in the removal of approximately 30% of the protein. A significant fraction of the residual complex is still stable in concentrated salt solutions, and can be dissociated by Mg(II). Extensive digestion with pronase results in the removal of the protein and the cleavage of the DNA chain.


Gene 4 protein of bacteriophage T7. Purification physical properties, and stimulation of T7 DNA polymerase during the elongation of polynucleotide chains. With the use of an in vitro complementation assay to measure activity, the gene 4 protein of bacteriophage T7 has been purified 1000-fold to yield a nearly homogeneous protein. The purified gene 4 protein is a single polypeptide having a molecular weight of 58,000. In addition to being essential for T7 DNA replication in vivo and in vitro, the gene 4 protein is required for DNA synthesis by the purified T7 DNA polymerase on duplex T7 DNA templates. In the absence of ribonucleoside 5'-triphosphates, DNA synthesis by the gene 4 protein and the T7 DNA polymerase is dependent on phosphodiester bond interruptions containing 3'-hydroxyl groups (nicks) in the duplex DNA. The reaction is specific for the T7 DNA polymerase, but any duplex DNA containing nicks can serve as template. The Km for nicks in the reaction is 3 x 10(-10) M.


Presence of adipose fat as a criterion of implant compatibility. An analysis of the tissue sections from previous implant studies was performed define additional criteria which could be considered in determinations of biocompatibility of implant materials. Adult albino rabbits were implanted with biomaterials in the sacrospinalis muscle for periods of 2, 6, 18, and 54 weeks. Fourteen different implant materials were used in this study. The tissues were examined histologically for the appearance of adipose fat cells within the membrane surrounding the implant as an important criterion of tissues implant compatibility. The results were compared with other previously used criteria in judging biocompatibility of implant materials. For the most compatible nonreactive materials, adipose tissue formation within the pseudomembrane began at 6 weeks and was quite extensive at 54 weeks. The reactive materials studied by us did not exhibit this phenomenon.


An approach to the soft tissue/synthetic material interface. It is argued that chemical modification of soft tissue implants in the hope of obtaining an associated tissue response is unlikely to succeed as a method for studying the fundamentals of implant/tissue interactions. An alternative approach is proposed which places greater emphasis on the interfacial interactions (such as protein adsorption) which occur after implantation, in a manner paralleling current advances in knowledge of the blood/material interface. From simple arguments, it is proposed that the observed similarities in soft tissue response of hydrophobic materials may result from irreversible protein adsorption, and that if unusual tissue responses are possible they are likely to be found only with hydrophilic implants. The possiblity of a critical hydrophilic/hydrophobic character which an implant must possess for essentially irreversible protein adsorption is also discussessed.


Intranuclear actin bundles induced by dimethyl sulfoxide in interphase nucleus of Dictyostelium. Electron microscopic evidence demonstrated that dimethyl sulfoxide (DMSO) induces formation of giant intranuclear microfilament bundles in the interphase nucleus of a cellular slime mold, Dictyostelium. These giant bundles are approximately giant bundles are approximately 3 micrometer long, 0.85 micrometer wide, and composed of microfilaments 6 nm in diameter. Studies in which glycerinated cells were used showed that these microfilaments bind rabbit skeletal muscle heavy meromyosin, forming typical decorated "arrowhead" structures, and that this binding can be reverted by Mg-adenosine triphosphate. These data verify that the intranuclear microfilaments are the contractile protein actin, and that DMSO affects intranuclear actin, inducing the formation of such giant bundles. The intranuclear actin bundles appear at any developmental stage in two different species of cellular slime molds after treatment with DMSO. The native form of the intranuclear actin molecules and their possible functions are discussed, and it is proposed that the contractile protein has essential functions in the cell nucleus.


Microtubule system of isolated fish melanophores as revealed by immunofluorescence microscopy. The microtubule system of melanophores of the angelfish, Pterophyllum scalare, has been studied using antibodies prepared against purified porcine brain tubulin in indirect immunofluorescence microscopy. Melanophores were freed from the surrounding tissue components of isolated scales by mild enzymatic digestion and then allowed to settle on a glass cover slip. In both the dispersed and the aggregated states large numbers of fluorescent fibers are seen. The number and the astral arrangement of these fibers, which run from the central region to the periphery of the cell, are striking. The system of fluorescent fibers is replaced by diffuse fluorescence of moderate intensity after cold treatment, but is restored after rewarming the cells. Differences in the immunofluorescence profiles between cells with dispersed and aggregated pigment are discussed in relation to electron microscopic data available for this system.


Phagocytosis of bacteria by polymorphonuclear leukocytes. A freeze-fracture, scanning electron microscope, and thin-section investigation of membrane structure. The changes in membrane structure of rabbit polymorphonuclear (PMN) leukocytes during bacterial phagocytosis was investigated with scanning electron microscope (SEM), thin-section, and freeze-fracture techniques. SEM observations of bacterial attachment sites showed the involvement of limited areas of PMN membrane surface (0.01-0.25mum(2)). Frequently, these areas of attachment were located on membrane extensions. The membrane extensions were present before, during, and after the engulfment of bacteria, but were diminished in size after bacterial engulfment. In general, the results obtained with SEM and thin-section techniques aided in the interpretation of the three-dimensional freeze-fracture replicas. Freeze-fracture results revealed the PMN leukocytes had two fracture faces as determined by the relative density of intramembranous particles (IMP). Membranous extensions of the plasma membrane, lysosomes, and phagocytic vacuoles contained IMP's with a distribution and density similar to those of the plasma membrane. During phagocytosis, IMPs within the plasma membrane did not undergo a massive aggregation. In fact, structural changes within the membranes were infrequent and localized to regions such as the attachment sites of bacteria, the fusion sites on the plasma membrane, and small scale changes in the phagocytic vacuole membrane during membrane fusion. During the formation of the phagocytic vacuole, the IMPs of the plasma membrane appeared to move in with the lipid bilayer while maintaining a distribution and density of IMPs similar to those of the plasma membranes. Occasionally, IMPs were aligned to linear arrays within phagocytic vacuole membranes. This alignment might be due to an interaction with linearly arranged motile structures on the side of the phagocytic vacuole membranes. IMP-free regions were observed after fusion of lysosomes with the phagocytic vacuoles or plasma membrane. These IMP-free areas probably represent sites where membrane fusion occurred between lysosomal membrane and phagocytic vacuole membrane or plasma membrane. Highly symmetrical patterns of IMPs were not observed during lysosomal membrane fusion.


Inhibition of mammalian ribonucleotide reductase by a dinucleotide produced in eucaryotic cells. HS3, a highly phosphorylated dinucleoside originally purified from the fungus Achlya, has been isolated from Chinese hamster ovary cells undergoing glutamine starvation. The HS3 compounds obtained from the fungal and mammalian sources exhibited similar physical and chemical properties. This unusual dinucleotide may be an important regulator of eucaryotic ribonucleoside diphosphate reductase activity; for 50 micrometer HS3, isolated from either mammalian or fungal cells, significantly inhibited CDP reduction in Achlya or hamster cell preparations, but only marginally affected the activity of the enzyme from E. coli. Studies with HS3 isolated from Achlya and partially purified mammalian ribonucleotide reductase indicated that the compound noncompetitively inhibited the reduction of varying concentrations of the substrates CDP, ADP and GDP with Ki values of 23 micrometer, 14 micron and 16 micron respectively. These inhibitor concentrations are well below the estimated intracellular levels of HS3 in glutamine starved cells and suggest that HS3 inhibition of ribonucleotide reduction may be responsible for the rapid inhibition of DNA synthesis seen under these culture conditions.


Distribution of anionic sites on surface of B cell granule and plasma membranes: a study using cationic ferritin. The distribution of anionic sites on the membranes of rat pancreatic B cells and of their storage granules has been studied by the use of a visual probe of cationic ferritin. Membranes of isolated storage granules possessed a net negative charge which was apparently evenly distributed; the number of anionic sites was not markedly altered by prior incubation of the granules with neuraminidase or with 10(-5) to 2 X 10(-3) M calcium chloride. Distribution of charges along B cell plasma membranes was less uniform but was similarly unaffected by alterations of calcium concentration, or by neuraminidase treatment. However, during the fusion of plasma membrane and granule membrane which occurs in exocytosis, the emerging granule membrane was found to be devoid of anionic sites. The implications of these findings for the regulation of insulin secretion by exocytosis are discussed.


[A tumour of the transverse mesocolon (author's transl)]. The authors report a case of mesenchymoma of the transverse mesocolon and analyse 50 cases of tumour of the transverse mesocolon found in the literature. They emphasise the diagnostic difficulties, the pathological characteristics and consider the surgical problems raised by these tumours.


Isolation of bacteriophages specific for the K1 polysaccharide antigen of Escherichia coli. Five bacteriophage stocks were prepared after enrichment of a sewage sample using Escherichia coli 02:K1:H4 (strain U9/41). The bacteriophages were tested for their ability to lyse 224 strains of E. coli that had been tested for the presence of the K1 antigen by means of an antiserum-agar diffusion technique, using a meningococcus group B antiserum known to detect the E. coli K1 antigen. The standard test strains for E. coli K antigens 2 to 99 were used as control strains. Of the 101 strains found to possess the K1 antigen using the antiserum-agar technique, 93 were lysed by at least one of the bacteriophages, whereas 8 of the 123 strains apparently lacking K1 were lysed by one or more of the bacteriophages. None of the standard test strains for K antigens 2 to 99 was lysed by any of the bacteriophages. The eight strains thought to lack K1 but that were lysed by bacteriophage were re-examined by immunoelectrophoresis, using meningococcus group B antiserum; five of the eight strains gave a precipitin line corresponding to K1. The use of K1-specific bacteriophages offers an inexpensive and easy method for the identification of the K1 antigen.


Evaluation of the oxi/ferm tube system with selected Gram-negative bacteria. The Oxi/Ferm test system was evaluated for accuracy and reliability for identification of nonfermentative and oxidase-positive fermentative bacteria by using 375 bacterial strains obtained from stock culture and clinical specimens. The Oxi/Ferm system is a compartmentalized tube containing eight media to provide nine biochemical test results. When combined with the oxidase test, the results corresponding to the positive reactions are totaled and the composite number is located in the coding manual to identify the organisms. The 375 isolates studied were evaluated for accuracy of identification, using both the original and revised code manuals. In comparison with the conventional media used, there was 100% correlation in tests for hydrogen sulfide and indole production, over 96% for nitrogen gas, arginine, and urease, over 92% for xylose and dextrose oxidation, and less than 90% for citrate utilization and dextrose fermentation. There was an overall accuracy in identification of 89.3% using the original manual, with accuracy revised slightly upward to 90.7% using the revised manual. There was 100% accuracy in identification with 44.0% of the strains tested (11 species) using the original manual and with 66.1% (16 species) using the revised manual. Thirteen of the 40 original misidentifications and 14 of 35 revised misidentifications resulted from failure to code and were unidentifiable by Oxi/Ferm. The remainder were incorrectly identified or could not be differentiated from closely related strains. Eleven strains of Alcaligenes odorans were correctly identified using the original code, whereas no code was provided in the revised manual. The Oxi/Ferm system is both simple and rapid and is satisfactory for identification of the more common isolates.


Effect of temperature on survival of Bacteroides fragilis subsp. fragilis and Escherichia coli in pus. Approximately 40 to 60% of Bacteroides fragilis subsp. fragilis in pus from experimental intra-abdominal abscesses lost their viability within 24 h when stored at refrigeration temperature (4 degrees C) either aerobically or anaerobically. No viability loss of B. fragilis was noted when pus was stored at 25 degrees C. Only slight loss of viaability of B. fragilis was observed at 15 degrees C. Escherichia coli coexisting in pus with B. fragilis increased several 100fold in 24 h when stored at 25 degrees C, but no significant growth occurred when they were kept at 15 degrees C. Approximately 20 to 40% of E. coli lost their viability when such pus was stored at 4 degrees C. We suggest that 15 degrees C may be an alternative temperature for storage of anaerobic specimens in laboratories where some delay in routine processing is unavoidable.


Evaluation of two rapid methods for identification of commonly encountered nonfermenting or oxidase-positive, Gram-negative rods. The ability of the expanded API-20E and the Oxi-Ferm System to identify 176 isolates of nonfermenting or oxidase-positive, gram-negative rods from 17 species or groups was studied. The expanded API-20E, without referral to a computer reference center, was able to identify 61.4% of the isolates to the species level. If reference to the computer center was utilized, an additional 25% could be identified. Of the isolates examined, 13.6% were misidentified, partially identified, or assigned no identification. Those assigned to the incorrect genus constituted 0.5% of the isolates tested; those assigned to the correct genus, but the wrong species, constituted 0.5%; 4.5% were placed in the correct genus with no species given; and 7.9% were assigned no identification. The Oxi-Ferm System was able to assign 75% of the isolates to the correct species without further testing, and an additional 19.3% required additional testing for correct identification. Those assigned to the incorrect genus represented 3.4% of the isolates tested; 1.1% were assinged to the correct genus, but the wrong species; and 1.1% were assigned to the correct genus, with no species indicated.


API computer profiles: correlation of API 20E with API 10S. A comparison of identifications of 201 clinical isolates with the 21-test API 20 Enteric kit and the subset of tests in API 10S indicated 85.6% agreement at the species level and 93.5% agreement at the genus level.


His-Purkinje conduction during retrograde stress. The pattern of retrograde His-Purkinje conduction was evaluated in 28 patients using ventricular extrastimuli. In each patient progressive prolongations of His-Purkinje conduction (S2H2) which appeared as ventricular extrastimuli were induced at closer coupling intervals (S1S2). There was an inverse linear relationship of S2H2 to S1S2 which was cycle length-dependent: i.e., at any S1S2 interval the resultant S2H2 was less at shorter drive cycle lengths. The degree of S2H2 delay varied widely (from 30 to 340 ms) and was unrelated to the presence of bundle branch block, H-V intervals, or capability of ventriculoatrial conduction. Prolongation of S2H2 was independent of intraventricular (muscle) conduction delay; such delay was usually absent at most, and occasionally all, S1S2 coupling intervals during which S2H2 was lengthening. Furthermore, in two patients both left and right ventricles were activated before the timed depolarization of the His bundle occurred, demonstrating that under the stress of extrastimuli, the impulse conducts through ventricular muscle with less delay than through the His-Purkinje system. We conclude that the His-Purkinje system typically displays slow conduction response to ventricular stress. The site of this conduction delay is probably at the distal "gate".


Effects of dihydroxy bile acids and hydroxy fatty acids on the absorption of oleic acid in the human jejunum. Perfusion studies of the normal human jejunum were performed to test whether dihydroxy bile acids and hydroxy fatty acids inhibit the absorption of oleic acid, since previous reports documented their inhibitory effects on the absorption of several other organic solutes. 3 mM deoxycholate and 7 mM glycodeoxycholate inhibited the absorption of 3 mM oleic acid in isotonic micellar solutions while inducing net fluid secretion. Similarly, fractional absorption of oleic acid decreased in the presence of hydroxy fatty acids. However, only the changes induced by 2 mM ricinoleic acid could be distinguished from changes induced by an increase in total fatty acid concentration. Under all experimental conditions, close linear relationships existed between net water movement and fractional absorption of glucose, xylose, and fatty acids, as well as between the absorption rates of these solutes. In contrast, net fluid secretion induced by hypertonic D-mannitol (450 mosmol/liter) had no effect on solute absorption. Our data and observations in the literature do not allow formulation of a hypothesis which would adequately define all effects of dihydroxy bile acids and fatty acids on intestinal transport processes. The observations help explain the malabsorption of fat and other nutrients in patients with the blind loop syndrome.