Beta-cell recognition of stereoisomers of D-glucose. The ability of all eight D-aldohexose steroisomers to stimulate insulin release and biosynthesis was compared with their ability to serve as a metabolic substrate for isolated islets of Langerhans as judged by formation of lactate. Insulin release and synthesis were stimulated by glucose or mannose but not by allose, altrose, gulose, idose, galactose or talose. No potentiary effects of allose, altrose, gulose, idose, or talose were found on insulin release in the presence of 4 mmol/l glucose nor did these sugars inhibit insulin release in the presence of 20 mmol/l glucose. Lactate formation was increased above values found in the absence of added substrate by 20 mmol/l D-glucose or mannose, but not by allose, altrose, gulose, galactose or talose. The results support the substrate-site hypothesis for the recognition of sugars as stimuli of insulin release and synthesis.


Properties of interferon induced by purified protein derivative of tuberculin in mice sensitized with BCG or cell-wall skeleton of BCG. Mice sensitized with either BCG or cell-wall skeleton of BCG (BCG-CWS) produced interferon in blood after stimulation with specific antigen, purified protein derivative of tuberculin (PPD). Both BCG-infected normal (C57BL/6) and thymic nude (BALB/c, nu/nu) mice showed enhanced activity to produce interferon by stimulation with E. coli endotoxin. However, detectable interferon was not produced in athymic nude mice sensitized with BCG or BCG-CWS by stimulation with PPD. Immune-induced interferon (I-IF) produced by BCG-CWS and PPD in mice was different in biological and physicochemical properties from virus-induced interferon. I-IF showed about 100 times more potent L-cell growth inhibitory activity than virus-induced L-cell interferon (L-IF). Both I-IF and L-IF showed macrophage-activating activity, which renders resting macrphages cytotoxic to L1210 leukemia cells. Antiviral and macrophage-activating activity of interferon preparation was not separated physicochemically in this study.


Cytolytic action of 60-F derived from live hemolytic streptococci against Ehrlich carcinoma cells. The effect of 60-F, a fraction obtained by 0.5 approximately 0.6 saturation of ammonium sulfate of streptomycin-pretreated cell-free extract from live hemolytic streptococci (avirulent Su strain), on release of 51Cr from the 51Cr-labeled Ehrlich ascites carcinoma cells was studied with following results: a) 60-F was found to be highly effective in releasing 51Cr from 51Cr-labeled Ehrlich ascites carcinoma cells. b) Additionally, destructive picutres of the tumor cells contacted with 60-F in vitro was observed by phase-contrast microscopic examination. c) Indication was that the 51Cr-releasing assay method is also useful for the study of the direct cytolytic effect of 60-F.


[Chloroplast DNA cloning in Escherichia coli. II. The properties of the recombinant plasmids bearing the EcoRI fragments of pea chloroplast DNA and the cloning of the DNA sequences with rRNA genes]. Previously a method of selection of colicine-defective recombinant plasmids by mitomycin C was described. A series of recombinant plasmids (CPS) with various EcoRI-fragments of pea chloroplast DNA has been obtained. This paper describes some properties of cloned fragments replicated in Escherichia coli. The alkali stability of recombinant plasmid DNAs has been demonstrated, indicating the absence of ribonucleotides in their structure. Heterogeneity of chloroplast DNA in nucleotide composition was demonstrated using ultracentrifugation analysis of CPS-plasmid DNAs in CsCl-actinomycin D density gradient. Pea chloroplast rDNA was cloned in recombinant plasmids.


[Genetic effects of N-nitroso-N-methylurea on Saccharomyces cerevisiae yeasts. I. The influence of radiosensitivity mutations on lethal and recombinogenic effects]. Effect of mutations rad2 and rad54 in homozygous state on survival, mitotic segregation and crossing-over induced by NMU in yeast was studied. Mutation rad2 did not influence on these effects of NMU. The mutation rad54 increased sensitivity to the lethal effect, the frequencies of NMU-induced segregation and crossing-over were decreased in the strain rad54 rad54. The recombinogenic effect of NMU on yeast was lower than under the action of UV and gamma rays.


[Nature of the compatible inheritance of hybrid plasmid pAS8 and its deletion mutants with replicon ColE1]. Hybrid plasmid pAS8, that consists of RP4 and ColE1 replicons, is incompatible with RP4 but co-exists with ColE1 replicon. The deletion mutants of pAS8, that replicates under the control of ColE1 replicon only, are incompatible with ColE1 derivatives, although the copy number of pAS8 and its deletion mutants in the cell is the same (1-3 per the chromosome). Incompatibility effect of plasmids is expressed in a sharp decrease in transformant's yield during selection of incoming and resident plasmids markers. In this case incompatibility is a very fast process, that leads to the elimination of resident or incoming plasmid just before plating on the selective medium. On the base of negative control theory, the repressors yield, synthesized in the presence of 1-3 copies of the deletion mutant is enough for the expression of ColE1-specific incompatibility. This ColE1 incompatibility is probably connected with the functional activity of ColE1 replicon. When ColE1 factor replicates under the control of RP4 replicon the expression of ColE1-specific incompatibility does not occur. Possibly, the incompatibility effect takes place when pAS8 deletion mutants cause the synthesis of ColE1-specific repressor. Also, the replicons of ColE1 may competein the membrane attachment site.


[Expression of eukaryotic genes in Escherichia coli cells]. The paper presents a survey of literature concerned with the possibility of expression of plasmid-clones genes from eukaryotic organisms in bacteria cells. Studies on bacterial synthesis of somatostatin, human insulin, hormone of rat growth and proteins: chicken ovalbumin and mouse dihydrofolate reductase are discussed.


[Mechanism of plasmid ColEl and pMB-9 mobilization by plasmid F'lac+ in Escherichia coli K-12]. The genetic control and mechanism of mobilization of the non-conjugative plasmids ColE1 and pMB-9 by the conjugative plasmids was orived to be recA-independent process in contrast to the mobilization of the chromosomal marker pro. Acridine orange and ethidium bromide curing data together with the results of electrophoretic analysis of plasmid DNA suggest that the plasmids F' lac+ and pMB-9 as well as F' lac+ and ColE1 remain autonomous after their contransfer to recipient cells. These data argue in favour of non-recombinational nature of the plasmid mobilization process. The possibility of transmission of a non-conjugative plasmid without transmission of a conjugative one from the donor strain carrying both plasmids was established. The results obtained are discussed with respect to the hypothesis on the effect of diffusible products encoded by the conjugative plasmid and required of the mobilization of the non-conjugative plasmid.


[Dependence of the viability and mutability of Escherichia coli K-12 cells on the balance of DNA and protein syntheses. II. The effect of disturbance in the balance of DNA and protein syntheses on the cellular mutability and viability of rec+ and rec- strains treated with nalidixic acid]. The changes in the survival and mutagenesis of rec+ and rec- Escherichia coli K-12 strains, treated with the selective inhibitor of DNA synthesis, nalidixic acid, are found to be due to the processes of the stabilization and repair of the metabolic gaps in DNA chains, which depend on the balance of DNA and protein synthesis. The various character is observed of the relation between the survival and the mutagenesis and the balance of DNA and protein syntheses in cells which are valuable (rec+) and defective (rec-) for the processes of DNA repair.


Replication in Saccharomyces cerevisiae of plasmid pBR313 carrying DNA from the yeast trpl region. Plasmid pBR313 carrying a 1.4 kb EcoRI fragment from the yeast TRP1 region (designated pLC544) is capable of transforming yeast trp1 mutants to Trp+ at high frequency (10(3)--10(4) transformants/micrograms DNA). Transformation can be achieved either by using purified plasmid DNA or by fusion of yeast spheroplasts with partially lysed Escherichia coli [pLC544] protoplast preparations. The Trp+ yeast transformants are highly unstable, segregating Trp- cells at frequencies of 0.18 per cell per generation (haploids) and 0.056 per cell per generation (diploids) in media containing tryptophan. Plasmid pLC544 replicates autonomously in the nucleus of yeast cells and segregation of Trp-cells is associated with the complete loss of plasmid sequences. In genetic crosses, pLC544 is randomly assorted during meiosis and is carried unchanged through the mating process into haploid recombinants.


Growth of Clethrionomys gapperi and Microtus pennsylvanicus in captivity. Growth of Clethrionomys gapperi and Microtus pennsylvanicus from southern Manitoba was documented from birth to 30 days. Clethrionomys gapperi were smaller and grew more slowly (g/day) than M. pennsylvanicus, although both species were weaned at the same age. There were significant differences in growth rates between sexes in M. pennsylvanicus and among litter sizes in both species. Most of the variability in growth was due to differences among litters. The patterns of development was similar to those reported for other North American species of Clethrionomys and Microtus.


Surface properties of cultured endothelial cells. Vascular endothelial cells (EC) are non-thrombogenic. The basis for this property is still essentially unknown. This paper summarizes our previous studies on EC associated glycosaminoglycans and on the specific binding of heparin to EC and describes the preventive effect of heparin sulphate (HS) on platelet adhesion to collagen; attempts to affect the binding of platelets to EC by specific removal of endogenous HS were, however, unsuccessful. The possible importance of EC surface HS for interactions with coagulation factors is discussed.


Platelet-reactivity of isolated constituents of the blood vessel wall. Collagens I and III, in fibrillar form, bound platelets equally well; both readily induced platelet aggregation. In contrast, collagens IV and V, although pretreated as collagens I and III to induce fibrillogenesis, failed to produce aggregation. No binding of platelets was detected. Lens capsule, containing collagen type IV in situ, was also inactive. Inactivity appears due to the lack of an appropriate quaternary structure since segment-long-spacing (SLS) aggregates of collagens IV and V, as of type I, induced aggregation. Elastin and its associated microfibrillar element did not aggregate platelets; some binding of platelets to elastin only was observed.


Platelet interactions with the endothelium and the subendothelium: the role of thrombin and prostacyclin. The adherence of 51Cr-labeled platelets to rabbit aortae everted on probes rotated in platelet-red cell suspensions has been measured. Platelet adherence to the subendothelium exposed by passage of a balloon catheter before everting the aortae was inhibited by compounds that increase platelet cyclic AMP levels (PGE1, PGI2 or dipyridamole). These agents, however, did not abolish platelet adherence to the subendothelium. Aspirin treatment of the vessel wall was used to block PGI2 production; platelet adherence to the surface of the 'undamaged' aorta and the subendothelium was studied following this treatment. Since aspirin treatment of the 'undamaged' vessel wall did not cause platelets to adhere to it, it seems unlikely that PGI2 formation by the vessel wall is the mechanism that prevents platelet adherence to normal endothelium. In addition, PGI2 formation by the vessel wall does not appear to influence platelet adherence to the subendothelium, since adherence was not increased by aspirin treatment of the damaged wall. Thrombin treatment of the 'undamaged' vessel wall increased platelet adherence to the surface, but the adherent platelets were seen to be adherent only to small areas where the endothelium was lost or damaged. Heparin reversed the effect of thrombin. Similar results were found when the subendothelium was exposed to thrombin or thrombin and heparin.


Clot-promoting effect of platelet-vessel wall interaction: influence of dietary fats and relation to arterial thrombus formation in rats. A small piece of vascular tissue punched from a rat aorta is able to clot plasma. This coagulation process is promoted by blood platelets, especially after their activation. Thrombin, generated by this clotting process, plays a key role in vessel-wall induced platelet activation. Vascular prostacyclin inhibits vessel-wall-induced clotting of platelet-rich plasma, possibly by inhibiting platelet activation. Type and amount of dietary fats were shown to influence vessel-wall-induced clotting via at least four different mechanisms, namely: by modifying vascular prostacyclin formation; by affecting the clotting potency of the vascular tissue per sec; by an effect on some platelet property, probably connected with platelet activation; by influencing a plasma factor. Each of these mechanisms, as well as the nature of vessel-wall-induced coagulation, requires further investigation.


Prostacyclin, thromboxane A2 interactions in haemostasis and thrombosis. Prostacyclin and thromboxane A2 are products of arachidonic acid which play a role in the regulation of haemostatic plug and thrombus formation. Aspirin inhibits the synthesis of both compounds but is more active in blocking TXA2 formation; based on this, aspirin is suggested to have an anti-thrombotic effect. Other possible approaches to the development of anti-thrombotic drugs are discussed.


Endogenous mechanisms which regulate prostacyclin release. When infused intravenously into anaesthetized cats angiotensin II (1--4 micrograms/kg) released into the circulation an unstable substance that caused de-aggregation of platelet clumps, relaxed a strip of bovine coronary artery, and its release was blocked by aspirin and indomethacin. Because of these characteristics this substance is likely to be prostacyclin. Catecholamines and phenylephrine did not induce the release of prostacyclin. It is suggested that a chemical modification of the molecule of angiotensin II may render a peptide with little hypertensive properties which will be an activator of prostacyclin biosynthesis.


Vascular prostacyclin and plasminogen activator activity in experimental and clinical conditions of disturbed haemostasis or thrombosis. In several experimental and clinical conditions associated with thrombotic tendency, or complicated by thrombotic episodes, either prostacyclin (which inhibits platelet aggregation) or plasminogen activator (which promotes fibrinolysis), or both, appear to be decreased in the vessel wall. In other conditions, however, either activity may not change or even be increased. Possibly, vascular damage is followed by an early stimulation of both activities (a defence mechanism?) which may be subsequently reduced or exhausted. While the role of vascular plasminogen activator in haemorrhagic conditions is apparently unknown, prostacyclin activity appears to be markedly enhanced both in experimental animals and in patients with uraemia and bleeding complications. There is a suggestive evidence that uraemic plasma powerfully stimulates vascular prostacyclin generation.


Antibiotic properties of lignin components. Inhibitory effects of compounds with guaiacyl and syringyl structure, representing the structure of native lignin, were studied on model cultures of bacteria, yeasts, yeast-like microorganisms and moulds. Isoeugenol exhibited the most pronounced inhibitory effect on growth of the studied microorganisms.


[Juvenile cadaver kidneys as suitable donor organs]. As several authors stated within the last few years and according to our experience as well cadaveric kidneys of young children are suitable for transplantation. Considering the shortage of transplantable organs en-bloc-transplantation should not be performed, even if this provides the opportunity to remove one kidney in case of surgical complications. On the other hand by transplanting a single organ one donor may help two recipients. Since it is known that kidneys of babies under 18 months do not guarantee sufficient function in adults, it should be a rule that kidneys of children under 18 months have to be transplanted en-bloc, whereas kidneys of older children can be transplanted solitarily without hesitation. Long term results with baby kidneys do in no way differ from kidneys of adults.


[Kidney preservation by mechanical perfusion or hypothermic storage]. The efficiency of hypothermic mechanical perfusion and hypothermic storage, resp., for kidney preservation was to be examined. For this purpose dog kidneys were subdued to 0 to 60 min of warm ischemia, then preserved for 12--72 hours and thereafter transplanted. It could be concluded: 1. Hypothermic mechanical perfusion makes a successful 72 hour preservation possible with excellent kidney function immediately after transplantation. After 30 minutes of warm ischemia the preservation period should be limited to 24 hours. 2. Hypothermic storage is inferior to mechanical perfusion concerning the immediate function after transplantation: 24 hours storage time and 15 minutes of warm ischemia should not be exceeded. 3. Kidney function decreases exponentially by the time of preservation. This means that the warm ischemic period and the preservation time, resp., should be as short as possible to get an undamaged kidney after transplantation: the shorter the preservation period the better the kidney function after transplantation.


[Kidney transplantation in children].

[Kidney transplantation in children]. Ten years' experience of renal transplantation in 40 children (aged 5--18 years) is reviewed. Including 3 second transplantations 40 cadaver grafts and 3 living donor grafts were transplanted. Mean graft survival was found to be 19 months, the longest being 8 years. At the time of last examination 24 children were alive wtith a functioning graft (mean serum creatinine level 1.5 mg/dl). After graft failure 9 children were again on intermittent hemodialysis. 7 children had died mainly because of infections or cardiovascular complications. Cumulative survival rate of patients and cadaver grafts after 2 years was 84 and 53% respectively. Growth velocity was variable but improved in the majority of patients compared to the previous dialysis period. Full social rehabilitation could be achieved in every case. Main complications were acute rejections (irreversible in 7 of 67), chronic rejections (irreversible in 6 of 10), side effects of immunosuppressive drugs (infections; hematologic, metabolic, and bone disorders), hypertension (60%), recurrence of primary renal disease (in 5 of 9 patients with focal segmental glomerulosclerosis) and surgical complications (28%). Therapeutic guide-lines are given emphasizing the role of specialized children's centres and the cooperation between pediatric nephrologists, immunologists, urologists and psychologists including the time before, during and after transplantation.


[Therapy of Amanita phalloides poisoning. Results of clinical and experimental studies]. On the basis of experimental and clinical results evidence accumulates that supportive therapy is still the milestone in the therapy of death cap poisoning. Today the letality is 20%. The main cause of the intoxication are the amatoxines which inhibit DNA dependent RNA Polymerase II or B. Parts of the supportive therapy are: diuresis with 150--200 ml urin/hour, careful corrections of disturbances in electrolyte and acid-base-metabolism, oral administration of charcoal and oral gut sterilization. In spite of a multitude of experimental and clinical reports on the effectivity of Penicillin G, thiocticacid and steroids there are no controlled studies to demonstrate the advantage of one regimen over the other. The time course of resorption and excretion of amatoxines clearly shows that hemoperfusion and/or hemodialysis are only of value if applied within the first 24 hours after poisoning. At this time neither anamnestic nor biochemical data give any clues to the probable course of the disease. Heterologous baboon liver perfusion may be lifesaving in coma hepaticum grade IV, but the small amount of cases so far does not yet allow any comments on its effectivity.


Demonstration of calcitonin and carcinoembryonic antigen (CEA) in medullary carcinoma of the thyroid (MCT) by immunoperoxidase technique. The presence of calcitonin and of carcinoembryonic antigen (CEA) was studied in six cases of medullary carcinoma of the thyroid using an immunoperoxidase technique. In five cases the material was obtained surgically and in one at autopsy. Tissue from primary tumours as well as from metastases was studied. Calcitonin and CEA were identified within all the tumours studied, although their pattern of distribution and staining intensity varied both within the cells and within the tumour. Some parts of the tumour contained both CEA and calcitonin, while others stained positively only for one of these substances. In some parts of the tumour there was no positive staining for either substance. Within the cells, CEA showed a typical linear distribution along cell surfaces, while calcitonin showed a more even cytoplasmic distribution and the deposits were more granular. Normal tissue surrounding tumour deposits did not show positive staining. It is considered that cells of medullary carcinoma of the thyroid contain both calcitonin and CEA. Identification of CEA and calcitonin in tumour tissue can be used as a diagnostic aid to identify medullary carcinoma of the thyroid. Iit is considered that these substances are being produced by this tumour and can be used as tumour markers.


Legionnaires' disease: case report with special emphasis on electron microscopy and potential risk of infection at autopsy. A case of Legionnaires' disease (LD) is presented in which the diagnosis was made initially by EM and later confirmed by specific immunofluorescence of formalin-fixed, paraffin-embedded lung tissue. The possibility of an infection with Legionella pneumophila during autopsy is suggested by the fact that the pathologist who performed the post-mortem examination developed signs and symptoms of an upper respiratory tract infection 2 weeks afterwards. Approximately 4 weeks after the onset of his illness a significant antibody titre against L. pneumophila was found in his serum, whereas 6 months later the antibody titre was insignificant.


The neuroendocrinology of male reproduction. Androgen biosynthesis occurs in Leydig cells and spermatogenesis in the seminiferous tubules, but both are under control of the anterior pituitary through its secretion of gonadotropins. The hypothalamus is primarly involved in feedback control of gonadotropin secretion in response to varying blood levels of testicular hormones. Another pituitary steroid, prolactin, also is involved in male behavior.


Differentiation of functionally active mouse T lymphocytes from functionally inactive bone marrow precursors. IV. Recovery of T-cell function from bone marrow precursors in a histo-incompatible environment. Regeneration of T-cell activities in vivo or in vitro from mouse bone marrow precursors differentiating in the presence of an allogeneic thymus was investigated. The data indicated that T-depleted bone marrow cells fail to affect long-term reconstitution of allogeneic recipients unless a pool of rapidly maturing T-precursor cells is also removed (post-thymic pool). Animals reconstituted with pre-thymic bone marrow stem cells become stable chimaeras in which cells capable of generating an in vitro CML response to host antigens, as well as cells capable of suppressing that response, could be demonstrated. Similar data (CML directed against the H-2 antigens of the 'host' thymus feeder layer and cells capable of inhibiting that response) were obtained when pre-thymic bone marrow cells were grown in vitro on allogeneic thymus feeder cells. When cytotoxic T-lymphocyte precursor (CTLp) and helper (CTLh) cells were separately investigated, a restriction in their co-operation for an anti-host response was observed when precursor cells differentiated in an allogeneic environment. Only CTLp and CTLh differentiating in the presence of the same allogeneic thymus source (whether in vivo or in vitro) could co-operate to generate CTL directed to H-2 antigens of that thymus source.


Studies on the control of antibody synthesis. XIII. Preferential depeletion of precursors of high affinity antibody-secreting cells by specific immunoadsorbents. Passage of spleen or bone marrow cells from non-immunized donors through a DNP-Ova coated Degalan bead immunoadsorbent column resulted in a specific depletion of the ability of the cell population to restore the anti-DNP antibody response to DNP-BGG by lethally irradiated mice. There was a preferential depletion of precursors of high affinity antibody-secreting cells which is consistent with the view that these cells bear high affinity antigen receptors. The cell population passed through the immunoadsorbent column was also preferentially depleted of the capacity to produce high affinity direct anti-DNP PFC in comparison with thigh affinity indirect anti-DNP PFC. These findings suggest, but do not prove, that different subpopulations of antigen-binding cells are precursors for direct and indirect PFC.


Peritoneal exudate T lymphocytes with specificity to sheep red blood cells. II. Inflammatory helper T cells and effector T cells in mice with delayed-type hypersensitivity and in suppressed mice. Peritoneal exudate cells were induced in mice 4 days after immunization with SRBC. A low dose of SRBC (10(6) i.v.) caused T lymphocytes to appear in inflammatory exudates. These cells, not only transferred DTH reactions, but also functioned as helper T cells in antibody production after transfer to syngeneic nu/nu recipient mice. After a high dose of SRBC (10(9) i.v.), very few helper T cells and no DTH transferring T cells were found in inflammatory exudates, although they were present in the spleen. It is postulated that T cells mediating DTH reactions and helper T cells behave similarly as far as those dose dependency of appearance in inflammatory exudates is concerned. A high dose of sensitizing antigen causes retention of helper and effector T cells in the spleen, in this way favouring antibody formation; low doses of antigen allow them to leave the spleen, thus favouring mediation of DTH reactions in the periphery.


Delayed rejection of cardiac xenografts in C6-deficient rabbits. Puppy hearts were engrafted to C6-deficient rabbits, and also to complement-sufficient animals in order to determine the influence of the sixth component of complement on xenograft rejection. Delayed rejection was observed in the puppy hearts engrafted to the C6-deficient animals indicating that complement sufficiency of C6 is required for hyperacute xenograft rejection.


The influence of T cells on the initiation and expression of immunological memory. Nude and normal CBA mice have been used in adoptive transfer experiments to analyse the development of immunological memory. The development of B-cell memory to xenogeneic erythrocyte antigens is to a very large degree dependent on the presence of T cells, with IgG memory being somewhat more dependent than IgM memory. In this system, the expression of B memory, that is the transformation of memory cells to antibody-secreting cells under the inductive influence of antigen, is largely dependent on the presence of T cells. Primed (educated) T cells can have an antigen-specific potentiating effect on unprimed B cells in the presence of antigen.


Studies on B-cell memory. II. T-cell independent antigen can induce B-cell memory. Both athymic nude mice and normal mice primed with a T-cell independent antigen, i.e. dinitrophenylated dextran (DNP-DE), at a sub-immunogenic dose, produced very poor anti-DNP responses to a later challenge with the same antigen. B-cell memory was expressed, however, as an enhanced IgM response after the challenge of the DNP-DE-primed mice with the T-cell dependent antigen (dinitrophenylated haemocyanin, DNP-KLH) in the presence of functional T-cells. Moreover, DNP-DE-primed spleen cells also revealed an enhanced IgM response after adoptive transfer into irradiated recipients and challenge with DNP-DE. The injections of DNP-DE-primed nude mouse serum into unprimed mice resulted in the reduction of anti-DNP response to the immunization with DNP-DE. These results indicate that (a) T-cell independent DNP-DE causes the differentiation of B cells not only into antibody-forming cells but also into memory cells, (b) these memory cells can be triggered in situ by the T-cell dependent DNP-KLH in the presence of helper T cells but not by T-cell independent antigen, and (c) some humoral factor(s) induced by DNP-DE-priming seems to interfere with the expression of B-cell memory only when challenged with T-cell independent DNP-DE.


Interaction of subpopulations of murine lymph node lymphocytes in antigen-induced [14C]-thymidine incorporation: T and B cell synergy in the response to antigen. The antigen-induced [14C]-thymidine incorporation of murine lymph node cells (LNC) that were non-adherent (NAD) or adherent (AD) to nylon wool was studied. In contrast to NAD-LNC, AD-LNC responded like unfractionated LNC, and these responses were T lymphocyte dependent. By co-culturing NAD-LNC with subpopulations of AD-LNC the cellular requirements and interactions necessary for maximal incorporation of [14C]-thymidine were determined. A synergistic effect was observed when NAD-LNC and AD-LNC were co-cultured. Synergism was not dependent on T lymphocytes or macrophages in the AD-LNC population but was associated with the B lymphocyte subpopulation. These results indicate that the number of B lymphocytes present in a population of LNC can significantly influence the magnitude of the response to antigen.


The effect of anti-microvillus membrane antibodies on the transport of IgG across the suckling rat intestine. Rabbits were immunized with a preparation of microvillus membranes derived from the absorptive cells of the small intestine of suckling rats. The resulting immune serum contained antibodies which reacted specifically with the brush borders and walls of cytoplasmic vesicles of the intestinal cells, and was more effective than normal serum in preventing the transport of IgG across the intestinal cells. These results reflect the participation of microvillus membrane components, or receptors, in the transport mechanism.


Human Ia-like antigens in non-lymphoid organs. Human Ia-like antigens in liver and kidney were shown by the immunofluorescence assay to be present mostly in the endothelial-mesenchymal cells of these organs. The parenchymal cells apparently contained no human Ia-like antigens. The antigens in liver and kidney were purified and shown to have the same subunit structure as human Ia-like antigens of cultured B-lymphoid cells. The human Ia-like antigens in non-lymphoid organs, not only in liver and kidney but also in testis, heart, muscle and brain, carried all the xenoantigenic characteristics of human Ia-like antigens expressed on lymphoid cells of B-cell lineage.


In vitro investigation of autoantibody-secreting peritoneal cells and their regulation. A high proportion of peritoneal cells from untreated mice, after 4--5 days in culture, develop into plaque-forming cells against bromelain-treated mouse red blood cells. The number of plaque-forming cells was increased significantly by exposing the peritoneal cells to ammonium chloride to lyse red blood cells before culture. Conversely, the increase was significantly inhibited by adding before culture untreated or bromelain-treated sheep or mouse red blood cells. Treated or untreated horse or rat red blood cells did not inhibit the increase. Treating peritoneal cells or subpopulations of peritoneal cells with anti-theta serum and complement before culture caused a significant increase in the number of plaque-forming cells against bromelain-treated red blood cells after 3--4 days of culture. Various procedures were used to fractionate peritoneal cells into B-cell enriched and B-cell depleted subpopulations before culture and after culture, to investigate whether some of the plaque-forming cells could be attributed to phagocytic cells. Generally, changes in the number of plaque-forming cells against bromelain-treated mouse red blood cells paralleled changes in B-cells. In some experiments the proportion of plaque-forming cells observed represented up to 85% of the B-cells present. The results suggest that the high level of autoreactivity is due to antibody production by B-cells and that phagocytic cells are not forming spurious plaques. Further, it appears that the autoimmunity is regulated by T-cells and can also be inhibited by mouse RBC.


Secretory IgA does not enhance the bacteriostatic effects of iron-binding or vitamin B12-binding proteins in human colostrum. Human milk contains an unsaturated iron-binding protein (lactoferrin) and an unsaturated vitamin B12-binding protein. Lactoferrin has bacteriostatic properties, and a bacteriostatic role for the B12-binding protein has been postulated. In this study the bacteriostatic effect of lactoferrin was confirmed for strains of Escherichia coli, Pseudomonas and Proteus. Growth inhibition attributable to the unsaturated B12-binding protein could be demonstrated only with a known vitamin B12-dependent E. coli. It has previously been shown that the bacteriostatic effect of lactoferrin is potentiated by horse IgG antibody, and a similar potentiating effect of secretory IgA antibody in colostrum and milk would have obvious importance. An attempt was therefore made to demonstrate potentiation of bacteriostatic effects by naturally occurring secretory IgA antibody to E. coli. The results obtained indicate that secretory IgA antibody does not enhance the growth-inhibiting effects of either lactoferrin or the vitamin B12-binding protein.


Long-term human peripheral blood monocyte cultures: establishment, metabolism and morphology of primary human monocyte-macrophage cell cultures. Human peripheral blood monocytes were maintained in in vitro culture for periods up to 4 months using a non-human serum source. Monocytes were cultured in Dulbecco's modified Eagle's medium buffered with 20 mM HEPES and containing 10% horse serum and 10% foetal calf serum. The metabolic and morphological changes which occur in vitro were investigated using microtitre, Linbro and T 25 cultures. During culture, monocytes increased in size, had increased membrane activity as visualized by SEM, and differentiated into a morphologically heterogeneous population of fusiform and epithelioid shapes. These cell types retained the ability to phagocytose E glut and EA and to rosette with EA and EAC. Larger giant polynucleated cells were also observed during culture; many of these lacked the ability to bind or phagocytose inert or antibody-coated erythrocytes. Increases in lysozyme release and acid phosphatase activity also occurred during culture. Cultured monocytes exhibited characteristic profiles of leucine and uridine uptake with maximal activity observed by 5 days of culture. There was no detectable uptake of thymidine. Detailed analysis of regulatory processes involved in monocyte growth and differentiation could be performed with this in vitro system.


Hydrocortisone and the antibody response in mice. II. Correlations between serum and antibody and PFC in thymus, spleen, marrow and lymph nodes. The discovery that corticosteroids can alter leucocyte distribution implies that suppression of PFC in the spleen is not necessarily indicative of suppression of the antibody response as a whole. Therefore we have examined IgM and IgG serum antibodies and PFC in the thymus, spleen, femoral marrow, popliteal, thoracic and mesenteric lymph nodes of mice given a single injection of hydrocortisone acetate at various times relative to primary immunization with SRBC. Contrary to certain reports, there was close correlation between suppression of serum antibody and of splenic PFC, from which it was predicted and verified that few PFC could be detected elsewhere. In our hands, therefore, splenic PFC measurements did give an accurate indication of suppression overall. The characteristics of suppression suggest some of the underlying causes, and these are discussed.


Immune response to extracellular and somatic antigens in streptococcal infection and sequelae. The aim of the present study is to define that temporal relationships of the IgM and IgG responses to streptococcal group A carbohydrate (CHO) in rabbits and in man. Rabbits were immunized with group A streptococci and the development of anti-group A carbohydrate (ACHO) was studied. ACHO which appeared one week after the beginning of immunization belonged to the 19S class of immunoglobulins (IgM). A two- to four-fold rise in ACHO titers and immunoglobulins of the 7S class (IgG) were observed after two weeks. Three weeks after the beginning of immunization, the ACHO titer was at a maximum. In the following months no further rises in titer were seen, and the antibodies belonged mostly to the IgG class. IgM and IgG responses to streptococcal CHO and to extracellular antigens in patients with pharyngitis, acute rheumatic fever (ARF), and acute glomerulonephritis (AGN) were studied. Higher values of IgM were found in pharyngitis and AGN sera than in ARF sera, probably reflecting the interval between streprococcal infection and time of bleeding. ACHO antibodies persisted in patients' sera for long periods and belonged to IgG and IgM. This suggests a continuous, rather than a persistent, production of ACHO.


Effect of antibodies and complement on the interaction between Escherichia coli 0111:B4 and polymorphonuclear leukocytes. The interaction of Escherichia coli 0111:B4 with polymorphonuclear leukocytes in the presence of specific antibodies and complement was studied. This strain, which is resistant to phagocytosis by polymorphonuclear leukocytes, may be ingested and killed by the phagocytes in the presence of both antibodies and fresh serum. The ineffectiveness of fresh serum to promote ingestion of E. coli 0111:B4 by the phagocytes in the absence of antibodies reflects the inability of this strain to activate the complement system through the alternative pathway. Investigation of the mechanisms of the bacterial killing by polymorphonuclear leukocytes showed that both antibodies and complement were required for the oxygen-independent bactericidal system, whereas they were not needed for the oxygen-dependent system.


[Meningitis in an infant caused by Pasteurella multocida. Case report and review of other manifestations (author's transl)]. The authors report on an infant with meningitis caused by Pasteurella multocida. The problems of aetiology, diagnosis, and therapy of these infections are discussed with reference to this patient and other cases of P. multocida infection.


[Antibacterial activity of cefotaxim in comparison with seven cephalosporins]. The antibacterial activity of cefotaxim and seven cephalosporins was determined in 1,112 fresh isolates using the microdilution technique. All of the cephalosporins tested were ineffective against enterococci. Cefalotin was the most effective agent against Staphylococcus aureus. Cefaclor was superior to cephalexin against Escherichia coli and Proteus mirabilis, and thus inhibited 20% more Enterobacteriaceae. Cefazolin was as effective as cefamandole against E. coli. Cefuroxime and cefoxitin were almost equally effective against E. coli, Klebsiella and P. mirabilis; more than 95% of the strains were sensitive. Cefuroxime and also cefamandole were much more effective than cefoxitin against Citrobacter and Enterobacter. Cefoxitin on the other hand was superior against Serratia and indol-positive Proteus species. Cefotaxim was by far the most active cephalosporin against Enterobacteriaceae, only 2% of the strains being resistant. More than 90% of the strains of E. coli, Klebsiella, P. mirabilis and Serratia were sensitive to 1 mg/l, the lowest degree of activity being displayed against Enterobacter; 82% of the strains were inhibited by 16 mg/l. Cefotaxim was the only cephalosporin which showed appreciable activity against Pseudomonas.


In vitro comparison of synergism between cefamandole and gentamicin or tobramycin by the triple layer agar method with enzymatic inactivation. A synergistic effect was shown with gentamicin and tobramycin by means of a triple layer agar technique and enzymatic inactivation of cefamandole after only four hours' incubation. When the strain is sensitive to cefamandole and aminoglycosides, synergy is observed against all the strains studied (Staphylococcus aureus, Proteus, Klebsiella, Escherichia coli, Enterobacter, and Haemophilus influenzae). No significant difference was noted between the cefamandole-tobramycin and the cefamandole-gentamicin combinations when the microbial strains were sensitive to the three antibiotics.


In vitro investigations with fosfomycin on Mueller-Hinton agar with and without glucose-6-phosphate. Determination of the minimal inhibitory concentration of fosfomycin in the agar dilution test on Mueller-Hinton agar showed that the addition of glucose-6-phosphate to the nutrient medium potentiates the action of fosfomycin against Escherichia coli, Klebsiella, Enterobacter, Citrobacter and Staphylococcus aureus, sometimes by as much as 256-fold. Such a potentiation of action was not detectable with Serratia marcescens, the individual Proteus species, Pseudomonas aeruginosa or enterococci. Fosfomycin is very effective against most medically important bacterial species on Mueller-Hinton agar containing 25 micrograms/ml glucose-6-phosphate. Over 90% of the cultures of E. coli, Citrobacter, Enterobacter, S. marcescens,Proteus mirabilis, Proteus vulgaris, Proteus rettgeri, P. aeruginosa, S. aureus and enterococci examined were inhibited by less than or equal to 64 micrograms/ml fosfomycin.


The effect of trimethoprim-sulphonamide, trimethoprim and sulphonamide on the occurrence of resistant enterobacteriaceae in human intestinal flora. After the administration of various antimicrobial agents for chemotherapeutic purposes a general change in the intestinal flora of the patient is often observed. In contrast, the combination trimethoprim-sulphonamide causes in most cases a strong selective decrease in the number of Enterobacteriaceae organisms for treatment periods as long as four to 12 weeks. The results correspond to clinical experience with trimethoprim-sulphonamide therapy in urinary tract infection where reinfections with resistant organisms are the exception.


Synergistic activity of co-trimazine and co-trimoxazole in the urine. Co-trimazine (sulphadiazine, 410 mg + trimethoprim, 90 mg) is a new drug combination developed especially for use in the treatment of urinary tract infections. In cross-over experiments in volunteers receiving daily doses of co-trimazine (2 X 500 mg and 1 X 1000 mg), co-trimoxazole (2X960 mg), or nitrofurantoin (3X50 mg), the degree of antibacterial activity of co-trimazine in the urine was at least as high as that of co-trimoxazole and much higher and more consistent than that of nitrofurantoin. In further cross-over experiments in volunteers receiving co-trimazine 2X1000 mg or co-trimoxazole 2X960 mg for four days no or only slight activity was found in the urine against a sulphonamide-resistant Group D streptococcus, but distinct synergistic activity between the components was found against four Escherichia coli strains sensitive to trimethoprim and sensitive or resistant to sulphonamides. Against three of the latter strains the degree of activity in the urine was higher after co-trimazine than after cotrimoxazole. The synergistic action of trimethoprim and sulphonamide in the urine appeared greater with the former than with the latter combination.


Efficacy of two trimethoprim-sulphonamide combinations in experimental pyelonephritis in the rat. Co-trimazine, a new combination of trimethoprim and sulphadiazine intended for treatment of urinary tract infections, was tested and compared to co-trimoxazole in an experimental model of acute, obstructive pyelonephritis in the rat. The tests were carried out at four dose levels corresponding to the clinically recommended doses and to 1/4, 1/2, and 3/4 of the latter. The drugs were administered orally. Co-trimazine was found to have a greater antibacterial activity than co-trimoxazole and its use at a lower dosage than that of the latter appears warranted.


The pharmacokinetics of trimethoprim and trimethoprim/sulphonamide combinations, including penetration into body tissues. In this review the pharmacokinetic properties of trimethoprim (TMP) and TMP/sulphonamide combinations are discussed. The concentration of both substances in the various body fluids and tissues differ considerably from the concentrations in blood due to the different distribution of TMP and sulphonamides in the body. As a rule the ratio of the concentration of TMP to sulphonamide is higher in most body fluids and secretions than in blood. If inflammation is present, the concentrations vary considerably. The pharmacokinetic data must be taken into account when making in vitro tests. At the site of inflammation a TMP/sulphonamide concentration factor of 1:0.5 to 1:4 can be expected in urine and of 1:2 to 1:5 in tissue.


Development of sulphonamide-trimethoprim combinations for urinary tract infections. Part I: Comparison of the antibacterial effect of sulphonamides alone and in combination with trimethoprim. Plasma half life and in vitro activity were major criteria for selection of sulphonamides which are likely to give a strong synergistic action with trimethoprim in vivo. On the basis of literature data six sulphonamides, sulphadiazine, sulphachloropyridazine, sulphamethoxazole, sulphaisodimidine, sulphamerazine and sulphamethomidine appeared particularly suitable for combination with trimethoprim. An investigation of the activity in vitro of these compounds and their combinations with the latter against clinically isolated, sulphonamide-sensitive Klebsiella-Enterobacter and Escherichia coli strains showed optimal synergy at trimethoprim-sulphonamide ratios between 1:10 and 1:40, but that appreciable mutual potentiation occurred within a rather broad range of concentration ratios. Limited experiments indicated that synergy occurs less frequently and is less pronounced against sulphonamide resistant bacteria. The different sulphonamides behaved rather similarly in their combinations with trimethoprim, and in order to find the best sulphonamide, detailed comparisons of the pharmacokinetic properties of the different combinations are necessary.


Double-blind comparison of sulphonamide-trimethoprim combinations in acute uncomplicated urinary tract infections. The effects of a twice daily dosage of a combination of 410 mg sulphadiazine + 90 mg trimethoprim (SD + TMP) and 800 mg sulphamethoxazole + 160 mg trimethoprim (SMZ + TMP) were compared in uncomplicated urinary tract infections. All but one patient in each treatment group, i.e. 36 SD + TMP treated and 42 SMZ + TMP treated patients respectively, were cured. The percentage of side-effects related to therapy in the patients receiving the combination with sulphadiazine was 15.1% and in those with sulphamethoxazole 23.7%. Due to the small number tested, however, differences were not statistically different. It is noteworthy that only one of the SD + TMP patients had to stop therapy because of a rash, whereas therapy was stopped for this reason in three of the SMZ + TMP patients. SD + TMP represents a good alternative to SMZ + TMP in the treatment of urinary tract infections.


A clinical study of co-trimazine in comparison with co-trimoxazole and sulphalene in urinary tract infections. Co-trimoxazole, co-trimazine and sulphalene were tested in a controlled clinical trial of the treatment of urinary tract infections. Forty-three patients were treated with co-trimoxazole, 41 with co-trimazine, and 21 with sulphalene. During the study, sulphalene was found to be inferior to the other drugs and was excluded from further trials. The clinical efficacy of co-trimoxazole and co-trimazine was equal although the doses of trimethoprim and sulphonamide in co-trimoxazole were much higher than that in co-trimazine. In the latter sulphadiazine was used instead of sulphamethoxazole, the intention being to increase the concentration of active sulphonamide in the urine. Side effects tended to be less frequent in the patients treated with co-trimazine, although not significantly.


The use of co-trimazine and co-trimoxazole in elderly patients with urinary tract infections. In a double-blind study geriatric patients received either 410 mg sulphadiazine plus 90 mg trimethoprim (co-trimazine) or 800 mg sulphamethoxazole plus 160 mg trimethoprim (co-trimoxazole) twice daily. The investigation was carried out in order to compare the clinical efficacy and safety of co-trimazine in relation to co-trimoxazole with special emphasis on laboratory data in elderly patients suffering from uncomplicated urinary tract infections. Two to four weeks after start of treatment the original pathogen was eradicated in 80% of the 40 patients that were assessable for the effect of co-trimazine therapy, and in 77.8% of the 39 assessable patients in the co-trimoxazole group. There were no statistically significant differences between the two treatment groups in terms of laboratory data. Both drugs were well tolerated.


Clinical studies on co-trimazine in children. A study of co-trimazine tablets in 52 cases of urinary tract infection in children has shown that co-trimazine is at least as effective as co-trimoxazole, in which both the sulphonamide and the trimethoprim dose is higher than with co-trimazine. The frequency of side-effects (gastro-intestinal intolerance and allergic manifestations) was low. A sucrose-free suspension of cotrimazine for use in younger children was tested and found to have very good properties, as judged by a high acceptability rate among the young patients. Only one of 44 children refused to take the medicine during the prescribed fourteen days of therapy.


Clinical study of co-trimazine in urinary tract infections: a comparison with nitrofurantoin. Co-trimazine, a new preparation of sulphadiazine and trimethoprim, was compared with nitrofurantoin in a double-blind trial on patients with acute urinary tract infections. Co-trimazine had a bacteriological cure rate of 95% as compared to 88% for nitrofurantoin. There were no statistically significant differences in laboratory findings or in clinical side-effects between the two laboratory findings or in clinical side-effects between the two preparations. It was concluded that co-trimazine might constitute a valuable addition amongst drugs suitable for treatment of acute urinary tract infections.


Treatment of urinary tract infections with sulphonamide and/or trimethoprim. A preliminary report from a multipractice study. In a double-blind study 1194 patients with bacteriologically diagnosed urinary tract infection were randomly selected for treatment with either 1 g sulphamethizole twice daily, 200 mg trimethoprim twice daily, or 410 mg sulphadiazine plus 90 mg trimethoprim twice daily. All drugs were administered perorally for seven days. Two weeks after commencement of therapy 70% of the patients treated with sulphamethizole, 80% of those treated with trimethoprim and 85% of those treated with sulphadiazine/trimethoprim had sterile urine. After ten weeks 25% of the patients had bacteriuria irrespective of the form of treatment. Side-effects such as skin reactions occurred in 4.1% of the patients treated with trimethoprim alone, in 1.4% of those treated with sulphamethizole and in 3.2% of those treated with trimethoprim/sulphadiazine.


Influence of aminopenicillins on bacterial growth kinetics in vitro in comparison with the antibacterial effect in vivo. To quantify the relative antibacterial efficacy of beta-lactam antibiotics in vitro, a mathematical description of bacterial growth was developed. Curves of growth in the presence of the antibiotic are expressed as quadratic functions of time, with initial growth rate (ko) and rate of change of growth (a) as concentration dependant variables. For these variables the concentration-effect slopes were not parallel; therefore the potency ratio varied between 1 (at about 2 mg/l) and 2 (at higher concentrations). The relative efficacy in vivo was expressed as a potency ratio with regard to the number of bacteria in an infected thigh muscle in mice. The potency ratio was 1.5, which is between the limits of the in vitro results.


The aminopenicillins: development and comparative properties. In this review we have compared and contrasted the antibacterial and pharmacological properties of the aminopenicillins currently available. It is apparent that there is little hard evidence (except in the case of thphoid fever) that either amoxycillin or the pro-drug esters of ampicillin are significantly more effective than ampicillin itself, despite their better absorption and higher serum levels. However, amoxycillin and the ampicillin pro-drug esters do cause fewer bowel side-effects than ampicillin, which may justify their use despite the extra cost involved. The problem of susceptibility to penicillinases is unlikely to be solved by modification of the aminopenicillin molecule. A more likely solution is administration of these substances with beta-lactamase inhibitors such as clavulanic acid. Trials with such combinations are already under way.


Comparative in vivo activity of bacampicillin and amoxycillin. Bacampicillin is an orally well-absorbed pro-drug of ampicillin giving high in vivo levels of the latter. Its therapeutic activity was compared with that of amoxycillin in two experimental infection models in mice. The animals were infected with suspensions of Escherichia coli III and Haemophilus influenzae 22863 intraperitoneally and treated orally four hours afterwards with one of the two compounds. The antibacterial activity of the compounds was determined as CD50 values or by making viable counts in the blood and in organ homogenates of the animals. Ampicillin and amoxycillin had the same inhibitory but different bactericidal activity against the test strains. Both compounds appeared to have similar therapeutic activity and were found to cause a rapid decrease of the bacterial counts in the animals. Bacampicillin appeared to give a more rapid bactericidal activity than amoxycillin against the Haemophilus influenzae strain, whereas amoxycillin initially appeared more bactericidal against the Escherichia coli infection.


Efficacy of bacampicillin and ampicillin in experimental pyelonephritis in the rat. Bacampicillin and ampicillin were tested and compared with each other in a model of acute, obstructive pyelonephritis in the rat. The compounds were administered orally at five dose levels ranging from 50 mg/kg to 150 mg/kg. Bacampicillin was found to have a greater therapeutic activity than ampicillin, particulary at the higher doses, indicating that its improved absorption properties make it a therapeutically more effective compound than ampicillin.


Clinical pharmacological evaluation of aminopenicillins. To establish the clinical value of antibiotics, clinical trials are necessary. Both bacteriological and clinical-pharmacological data are helpful when designing a proper clinical trial. The difference between amoxicillin and ampicillin preparations cannot be defined on pharmacokinetic grounds alone. The difference in absorption between ampicillin and either pivampicillin or bacampicillin shows the latter two to be superior.


A dose response study with bacampicillin in uncomplicated gonorrhoea. In a systematic study of dose-response relationship 883 patients with uncomplicated gonorrhoea were given single oral doses of 400, 800 or 1600 mg bacampicillin, with 1 g probenecid. A single dose of bacampicillin 800 mg plus probenecid was the minimum effective dose in patients with fully ampicillin-sensitive strains. In patients with gonococci showing reduced sensitivity to ampicillin, 1600 mg bacampicillin was required to reach a cure rate above 95%. Bacampicillin was well tolerated in all treatment groups. Side-effects were reported in 4.6% of the courses of treatment. The most frequent adverse reaction was loose stools (1.9%), which was noted more often after the 1600 mg dose but was considered to be of no or very little clinical importance.


Bacampicillin and ampicillin in urinary tract infections: a double-blind comparison of efficacy and tolerance. A total of 100 hospitalized patients with upper and lower urinary tract infections were treated with bacampicillin or ampicillin. The treatment was double-blind and 50 patients were randomly allocated to each group, receiving 800 mg of bacampicillin or 2 g of ampicillin t. i. d. for ten days. The distribution by age and sex was similar in the two groups. Assessments of therapeutic response and tolerance were made two and six weeks after initiation of therapy. In 89% of the patients treated with bacampicillin and 68% of the ampicillin-treated patients both the bacteria and the symptoms were eliminated at the first follow-up visit three days after end of therapy. Relapse of reinfection occurred in 44% of the ampicillin-treated patients as compared to 7% in the bacampicillin-treated patients. Side-effects occurred significantly less frequently with bacampicillin compared with ampicillin (p less than 0.05). Treatment had to be stopped on account of side-effects in 16 patients treated with ampicillin compared with only four patients treated with bacampicillin.


Elimination of bacteria during antibacterial chemotherapy--a neglected parameter of chemotherapy. In 25 patients with urinary tract infections (pathogen Escherichia coli), colony counts from bacteria in the urine were done before and after treatment with antibiotics. Both 800 mg bacampicillin and ampicillin have a higher bactericidal effect in urine than co-trimoxazole. Patients with acute urinary tract infections given 400 mg bacampicillin and patients with chronic infections given double that dosage showed equally good results. In uncomplicated urinary tract infections with sensitive strains treated with cefotaxime, the colony count was reduced markedly faster than in infections in patients with kidney insufficiency or infection with multiresistant strains. The reduction of the colony count from the urine of patients with urinary tract infections and multiresistant strains during treatment with antibiotics is an important criterion in the evaluation of antibiotics. Such tests should supplement the customary sensitivity checks of antibiotics, the MIC and the agar diffusion test in the early phases of clinical trials.


A double-blind comparison of the clinical tolerance of bacampicillin and pivampicillin. A preliminary report. In this double-blind study, the clinical tolerance of high doses of bacampicillin hydrochloride (800 mg t. i. d.) and of pivampicillin base (700 mg t. i. d.) administered for ten days were compared in out-patients with acute urinary tract infections. The clinical response was comparable with that previously reported for those antibiotics. Upper gastrointestinal side-effects were more common after treatment with pivampicillin and were reported in 9/38 patients compared to 3/34 patients treated with bacampicillin. The duration of the symptoms of the side-effects was also more prolonged after pivampicillin than after bacampicillin.


Studies on aminopenicillin developments. Proceedings of a symposium. Concluding remarks. This symposium has focused on the bacteriological, pharmacokinetic, therapeutic, and adverse effects of the aminopenicillins. The compounds which are currently being assessed are primarily amoxycillin, ampicillin, bacampicillin, pivampicillin, and talampicillin. The three last mentioned are prodrugs which upon gastrointestinal absorption are rapidly split to yield circulating ampicillin. Pharmacokinetics and side-effects are reasons why one may narrow the selection of most suitable compounds within this group down to three. Ampicillin and amoxycillin are primary choices for parenteral dosage, and amoxycillin and bacampicillin the most suitable for oral therapy.


Conservative and prosthetic rehabilitation procedures in the dentition of old people. The therapeutic possibilities of conservative and prosthetic rehabilitation procedures are set out in this survey. In particular the aim was to find out whether these are suitable for retaining functional dentition into old age. The anatomical characteristics (Figs. 1 and 3) and secondary dysgnathic alterations of the dentition in the period of senescence make dental therapy considerably more difficult. The tooth-coloured filling materials do not behave as permanently in the mouth as could be wished. They are unsuitable for use in posterior teeth, and are in urgent need of improvement. The amalgam filling should still be regarded as the agent of choice for treatment of the cheek teeth, although cast precious metal inlays have been successful over a long period of time (Fig. 6). Too little use is made of veneer crowns in the anterior area with increasing age (Fig. 7), although the prospect of long term success is good. It was possible to establish statistically that tooth-coloured plastics are inferior to ceramic and metal ceramic crowns because of the greater retention of plaque. Ceramic should accordingly be preferred even with bridges in the anterior area. The replacement of missing teeth with removable partial dentures is the typical method of dental treatment for the period of senescence (Figs. 10-12). An attempt was made by bringing together various principles of planning and construction to lay down guidelines for dentures in older patients. With prosthetic care the fact that there are large differences in the physical and mental vitality of the older person, and that vitality frequently decreases rapidly for the most varied reasons, should not be overlooked. Even at this stage dentures should still prove functional.


The dental technician in Australia.

The dental technician in Australia. The training, employment and distribution of dental technicians in Australia is described. Since 1945 illegal practice by dental technicians has been widespread particularly among a group of technicians who operate one-man surgeries and operate illegally full time. Legislation has now been introduced in Tasmania, Victoria and New South Wales to permit registered technicians to make dentures direct to the patient and similar legislation is imminent in South Australia. The hope is expressed that the trend away from a technician being employed on the premises by the dental practitioner may be reversed, and that with the increasing sophistication of dental treatment and the technician being treated much more as a full member of the dental team, his interest and job satisfaction can be significantly improved.


In vitro adsorption of possible aetiological factors of hepatic encephalopathy. Four different adsorbents (activated charcoal, XAD-4, a strong base anion and a strong acid cation-exchange resin) were tested in vitro for their capacity to remove substances that may be important in the development of hepatic encephalopathy. Separate columns packed with one of these adsorbents were perfused for three hours with a reconstituted plasma solution containing simultaneously high concentrations of amino-acids, ammoniumchloride, short-chain fatty acids, octopamine and bile salts. Effective removal of all these substances was only obtained when either activated charcoal, or XAD-4, were combined with the cation-exchange resin. Possible implications for the treatment of hepatic coma are discussed.


Patient autonomy in alcohol rehabilitation. I. Literature review. A brief literature review on alcoholism treatment supports the authors' thesis that the alcohol abuser should be given some autonomy or "say so" in treatment selection. Presenting a variety of treatments as alternatives to the alcoholic may reduce treatment dropout rate, the aversive stigma frequently associated with therapy,and antiherapeutic disruptions while improving community coordination in alcoholism rehabilitation and providing an effective approach in treating the "total life" health of the alcoholic.


A comparative study with indomethacin and combined indomethacin sodium-salicylate in rheumatoid arthritis. A double-blind cross-over clinical trial was performed to compare clinical effectiveness of indomethacin (3 x 25 mg/day) alone to that of a combination of indomethacin + sodium-salycylate (3 x 25 mg/day and 3 x 250 mg/day, respectively) in rheumatoid arthritis. It was established that enteral blood loss was significantly reduced by combined treatment as determined by Cr51 labelled erythrocytes in comparison to that after treatment with indomethacin alone. Therapeutic effect was maintained in both groups, no significant disparities were observed. Occurrence of subjective complaints was less frequent in the combined treatment group. It was concluded that the combined preparation consisting of indomethacin and sodium-salicylate has a favourable effect in rheumatoid arthritis.


Efficacy of propranolol versus placebo in long-term treatment in patients with mitral valve prolapse. In 60 patients with mitral valve prolapse syndrome a randomized controlled interindividual double-blind study of propranolol (p) was performed. Patients received p 80 mg (A), 160 mg (B) and placebo (C) orally for 4 weeks. Prior to and after treatment, heart rate (HR) and blood pressure (RR) as well as systolic time intervals (STI) were measured and corrected for heart rate--electromechanical systole (QS2I), left ventricular ejection time (LVETI), and preejection period (PEPI). The ratio PEP/LVET was calculated. Plasma levels were measured by an optimized fluorimetric method. 1. STI lay in the upper part of the normal range, indicating that some patients had a hyperkinetic cardiac function. 2. P had no influence on QS2I and LVETI in A and B. PEPI was prolonged (A: +13.2 ms, B: +14.2 ms) and PEP/LVET was increased (A: +0.040, B: +0.050). 3. As indicated by the changes in HR, RR, PEPI, and PEP/LVET p showed in B compared to A only minor additional effects. 4. Plasma levels of p were in B three times higher than in A (A: 89.2 +/- 10.0 nmol/l B: 246.7 +/- 30.5 nmol/l). With a dose of 80 mg propranolol a point close to the plateau of maximum efficacy was reached, where a higher dose resulted only in small additional negative inotropic effects.


Sporotrichosis.

Sporotrichosis. Five cases of cutaneous sporotrichosis were seen at the Stanford University Department of Dermatology from 1966 to 1977, where the disease was previously unreported. The possibility of sporotrichosis should be considered when suppurative cutaneous lesions do not respond to antibacterial agents. Skin biopsy of sporotrichotic lesion is often nonspecific, while culture is the best method of diagnosis.


Parapsoriasis with hyper-IgE.

Parapsoriasis with hyper-IgE. A 56-year-old black man had generalized chronic dermatitis for two years before parapsoriasis en plaque (large type) with hyper-IgE (12,000-22,000 ng/ml) was diagnosed. Lymphocyte cultures showed decreased response to mitogens. Direct immunofluorescence of involved skin showed epidermal and dermal mast cells with bound IgE. Parapsoriasis with hyper-IgE may represent a form of "hypersensitive immunosurveillance" allowing a patient to reject a clone of malignant cells and retard transformation from a benign state (parapsoriasis) to a malignant state (mycosis fungoides).


Multicenter trial of fluocinonide in an emollient cream base. Working independently, eight investigators evaluated the clinical effectiveness of fluocinonide 0.05% in a three-week double-blind paired comparison trial of 240 patients. Half the investigators compared fluocinonide emollient cream with betamethasone valerate 0.1% cream in the treatment of atopic dermatitis, while the remaining four compared the study drug with triamcinolone acetonide 0.1% cream in the treatment of psoriasis. Statistical analysis showed clinical responses favored fluocinonide p = 0.021 (Wilcoxon Signed Rank) for the atopic dermatitis group and p = 0.001 (Wilcoxon Signed Rank) for the psoriasis group.


Congenital ichthyosis: concurrent immunodeficiency and atypical T cells. A patient with congenital ichthyosis and progressive neurologic anomalies showed disturbances in the specific humoral and cellular defense as well as the presence of atypical lymphoid cells in skin and lymph node. The latter resembled the atypical T cells found in mycosis fungoides and Sézary syndrome. The possibility of the presence of either a cutaneous T cell lymphoma or unregulated T cell stimulation leading to concurrent immunodeficiency in this patient is discussed.


Intervening briefly in the family system. A brief therapeutic intervention in a family is presented with the use of the Synallactic Collective Image Technique (S.C.I.T.). The technique proves to be a very useful tool for therapeutic intervention along General System Theory lines. The S.C.I.T. makes use of family paintings as a 2nd catalyst (the first one being the therapist) of transaction. Five sessions were held over a period of about one month. The family paintings reflect the successive phases through which the family transaction proceeded: From a dry, unsatisfying, mutually destructive relationship with no hope for co-operation and survival, through a phase of resistance during which each one was able to see his predicament, through a final phase after the 'storm' where the sun rises for the first time and co-operation for fruitful exploration begins. It has been observed that such a brief and intense intervention could lead, with proper handling, to the reactivation of self-leading processes, which generate more functional transaction within the family system.


Altered states of consciousness during psychotherapy: a historical and cultural perspective. This paper reviews the frequent phenomenon of altered states of consciousness in disparate cultural psychotherapeutic contexts. The historical antecedents of contemporary Western psychodynamic psychotherapy are examined and the central importance of altered states in the therapeutic effects of religious institutions such as the Dionysian rite and the Asclepia is illustrated. The continued presence of this phenomenon in Western psychotherapy from Mesmerism to psychoanalysis is shown. The use of trance states in the healing rituals of non-Western societies is culturally variegated therapeutic settings. The ubiquitous nature of the altered state phenomenon in such widely varied cultural contexts suggests the possibility of its being a universal component of psychotherapy.


A technique for quantitating enzyme histochemistry in adjuvant arthritic joints. I. Alkaline phosphatase. This paper establishes a quantitative technique for measuring the percentage of enzyme-reactive cells found in adjuvant arthritic articular cartilage, synovial membrane and bone marrow. Using alkaline phosphatase histochemistry and a recently-developed method of handling hard tissues, the technique is validated, the inherent variabilities determined and the existence of intra-articular lesions established. The technique now allows for a precise in situ evaluation of adjuvant arthritic drug inhibition by measuring enzyme-reactive cell alterations within the joint tissue most affected by the arthritis.


A technique for quantitating enzyme histochemistry in adjuvant arthritic joints. II. Acid phosphatase. This paper reports on the incorporation of acid phosphatase histochemistry with a quantitative technique designed to measure the percentage of histochemically-localized enzyme-reactive cells found in adjuvant arthritic articular cartilage, synovial membrane and bone marrow. With the data presented and the comparisons made, this incorporation is validated and adjuvant arthritic acid phosphatase "lesions" established. In addition to discussing the importance of the histochemical data and its relationship with intra-articular lysosomes, the applicability of the reactive cell values to drug inhibition studies is discussed.


Complement-mediated unspecific binding of immunoglobulins to some endocrine cells. Unspecific binding of immunoglobulins to gastrin G cells, glucagon A cells and somatostatin D cells of the gastric mucosa or pancreas, as well as to the calcitonin-somatostatin cells of rabbit thyroid has been found to occur through a non antigen-antibody mechanism mediated at least in part by the C1q fraction of complement. The phenomenon represents a major drawback in hormone immunohistochemistry, which can be prevented by incubating the specific anti-hormone sera with anti-C1q antibodies or with complement-fixing immunocomplexes.


Combined protein and DNA measurements by the ninhydrin-Schiff and Feulgen techniques. Feulgen nuclear staining with pararosanilin-SO2 was combined with the ninhydrin-Schiff technique. The aldehyde groups converted from primary amino groups are stained with an acriflavine-Schiff reaction. This results in a red nuclear fluorescence and a bright yellow cytoplasmic and nuclear fluorescence. The combined fluorescence staining facilitates cytofluorometric determination of total protein and DNA in the same cell. The ninhydrin-Schiff reaction is affected by the fixation procedure and the duration of the ninhydrin reaction. Investigations with a model system showed that proportionality between the fluorescence intensity of acriflavine and the amount of protein stained by the procedure was obtained after fixation with a fixation mixture suggested by Böhm et al. (1968) and a reaction with ninhydrin at 37 degrees C for 10 h. The ninhydrin-Schiff reaction has no effect on the fluorescence intensity of cells previously treated with pararosanilin-Feulgen staining and it is not affected itself by this previous procedure. Testing this double fluorescence staining on cytology specimens taken from patients with gastric carcinoma and uterine cervial carcinoma, cancer cells were shown to have markedly increased protein and DNA contents compared with those of normal cells.


Replacement of protamine by F1 histone during reactivation of fused human sperm nuclei. Rabbit antisera, specific for the histones F1, F2a2, F2b, F3 and for protamine were used to monitor a possible transition from protamine towards somatic-type histones during sperm nucleus reactivation, following human sperm fusion with mouse fibroblasts. Mature human sperm nuclei were shown to contain the histones F2a2, F2b, F3 and protamine, but were missing F1 histone by immuno cytochemistry using the indirect fluorescence method. However, a gradual disappearance of protamine from fused sperm nuclei, could be observed during the first 24 h of reactivation. Subsequently, F1 histone could be detected in increasing concentrations in 60% of reactivated sperm nuclei, during the next four days. The shift from protamine towards F1 histone could also be visualized cytochemically via staining with brilliant sulphaflavine, which appears to discriminate between sperm nuclei on the basis of their F1 histone content.


Immunoperoxidase reactions in resin embedded sections. A simple reproducible and reliable schedule for the immunoperoxidase demonstration of various substances in resin embedded sections is described. It has been applied to a variety of epithelial membrane antigens in carcinoma of the colon and to blood group substances. Preliminary work indicates that it may be applicable to most if not all of the antigens which survive conventional paraffin wax embedding.


Pathophysiology of pulmonary contusion in dogs. We produced a localized right lower lobe (RLL) contusion in 14 anesthetized ventilated dogs, 7 of which were treated with positive end-expiratory pressure (PEEP group). We measured gas exchange, pulmonary mechanics, and regional function before and 5 h after the contusion. Arterial PO2 decreased by 20 Torr and venous admixture doubled in both groups during air breathing. The shunt fraction (Qs/Qt) was minimally increased, despite a large lobar Qs/Qt (0.43) in the contused RLL. These results were explained by reduced ventilation per unit volume (VA/V), and ventilation-to-perfusion ratios of the contused RLL measured with 133Xe technique. We conclude that pulmonary contusion causes a leak of blood and plasma, flooding 25% of the air spaces of the RLL at FRC, reducing the compliance of adjacent air spaces, and resulting in a reduced VA/V and a large RLL Qs/Qt. These results are consistent with the observed reduction in regional volume and perfusion in the contused RLL, and suggest that Qs/Qt was not increased because blood flow was markedly reduced to flooded air spaces. PEEP reduced the hypoxemia, but increased the contusion.


Plasma volume during stress in man: osmolality and red cell volume. Our purpose was 1) to test the hypothesis that in man there is a range of plasma osmolality within which the red cell volume (RCV) and mean corpuscular volume (MCV) remain essentially constant and 2) to determine the upper limit of this range. During a variety of stresses--submaximal and maximal exercise, heat and altitude exposure, +Gz acceleration, and tilting--changes in plasma osmolality between -1 and +13 mosmol/kg resulted in essentially no change in the regression of percent change in plasma volume (PV) calculated from a change in hematocrit (Hct) on that calculated from a change in Hct + hemoglobin (Hb), i.e., the RCV and MCV were constant. Factors that do not influence RCV are the level of metabolism, heat exposure at rest, and short-term orthostasis (heat-to-foot acceleration). Factors that may influence RCV are exposure to high altitude and long-term orthostasis (head-up tilting). Factors that definitely influence RCV are prior dehydration and extended (greater than 2 h) periods of stress. Thus, either the Hct or the Hct + Hb equations can be used to calculate percent changes in PV under short-term (less than 2 h) periods of stress when the change in plasma osmolality is less than 13 mosmol/kg.


Spatial distribution of pulmonary blood flow in the dog with PEEP ventilation. The effects of mechanical ventilation without and with a positive end-expiratory pressure (PEEP) of 20 cmH2O and, after hemorrhage, on the spatial distribution of lobar lung blood flow were studied by means of iv injections of radioactive microspheres in supine dogs. The lungs were excised and dried at an inflation pressure of 30 cmH2O. Each lobe was cut into slices. In five slices from each lobe, 30--60 samples were punched out. The activity was measured in a gamma counter. By this means distribution of lobar blood flow could be described three-dimensionally. In dogs awake and spontaneously breathing or anesthetized and mechanically ventilated, blood flow distribution was rather even, except for a vertical gradient. PEEP caused a) a two- to threefold reduction in total lung blood flow, b) a redistribution from the right to the left lung, and c) a redistribution from the core to the periphery of the lung. This pattern of redistribution could not be reproduced by reducing cardiac output by hemorrhage during mechanical ventilation without PEEP.


Mechanism of action of monoketo-organomycin, cystaurimycin and their performic acid-oxidized modifications. I. Effects on bacterial growth and ribosomal peptidyl transferase activity. The response of 15 bacterial cultures to the inhibitory effects of the above mentioned compounds was investigated. These varied considerably. Escherichia coli cultures resistant to the compounds tested showed that monoketo-organomycin and cystaurimycin inhibit bacterial growth by what appears to  be the one and same mechanism which is not the same as that exhibited by their performic acid-oxidized modifications. This was confirmed by using cell free extracts of E. coli. In light of the puromycin reaction, using chloramphenicol and chlorotetracycline as control inhibitors, monoketo-organomycin and cystaurimycin were found to inhibit protein synthesis in vitro by inhibiting peptidyl transferase of ribosomes. In marked contrast, this enzyme was activated, as also was protein synthesis, by their performic acid-oxidized modifications. It was thus suggested that the growth inhibitory effects of the latter compounds might be due to their interference in other metabolic activities of the above test organism. The results obtained are discussed in light of the chemical similarities or differences existing between the compounds investigated.


Applications of self-control procedures by children: a review. Self-control procedures as used by children to affect their own behavior were reviewed. Particular emphasis was placed on self-instruction, self-determined criteria, self-assessment, and self-reinforcement. Self-punishment, comprehensive programs, and innovative self-control procedures (distraction and restatement of contingencies) were also evaluated. Basic effectiveness, comparisons with similar externally imposed interventions, maintenance, and the augmental value of the procedures were assessed. Important problems for future research were identified.


Specific inhibition of macrophage migration inhibition factor by fucosylated glycolipid RM. The effects of glycolipids on the interaction of the MIF (migration inhibition factor) with rat macrophages were examined using a migration inhibition assay system. MIF activity was specifically blocked by fucosylated Glycolipid RM [Gal alpha 1-3Gal(2-1 alpha Fuc) beta 1-3GalNAc beta 1-3Gal beta 1-4Glc beta 1-1ceramide, (1978) J. Biochem. 83, 85-90], but not by Cytolipin R, hematoside, or blood group B active glycolipid [Gal alpha 1-3Gal(2-1 alpha Fuc) beta 1-4GlcNAc beta 1-3Gal beta 1-4Glc beta 1-1ceramide]. Inhibition of MIF activity was proportional to the concentration of Glycolipid RM. These findings suggest that Glycolipid RM acts as a receptor for MIF.


A new surface treatment for bonding.

A new surface treatment for bonding. Bonding resins to enamel requires some form of mechanical attachment. Currently the dissolution of the outermost enamel layer is involved in etching with phosphoric acid. A new approach to mechanical bonding is described using a crystalline interface which itself bonds chemically to enamel. The crystals can be removed from the enamel with an ultrasonci or sickle-scaler followed by pumice prophylaxis. Many variations of the crystal growth principle may be visualized, including the development of fluoride-containing crystals.


Anterior fusion of the lumbar spine. End-result study with long-term follow-up. We reviewed the cases of fifty patients who underwent anterior lumbar-spine fusion with autogenous fibular and iliac-bone grafts and were followed for two to fifteen years. Their diagnoses were instability of the spine, degenerative disc disease, pseudarthrosis, and spondylolisthesis. Fifty-six per cent had union and 44 per cent, non-union. Those who had iliac grafts healed in an average of 2.5 years and those who had fibular grafts, in 5.2 years. The clinical result was successful in twenty-six patients (52 per cent) and unsuccessful in twenty-four patients (48 per cent). Paradoxically, about one-half of the patients with clinical successes had a non-union and one-half of the failures had union. Retrograde ejaculation (sterility) did not develop in any of the men, and a survey of world authorities on anterior spine fusion revealed only sixteen patients with the sequela of retrograde ejaculation. The incidence of that complication has been exaggerated.


Transiliac lengthening of the lower extremity. A modified innominate osteotomy for the treatment of postural imbalance. A modified innominate osteotomy was employed to treat postural imbalance by a hemipelvic lengthening in twenty patients, five to twenty years old. Diagnoses associated with the postural imbalance included acetabular dysplasia with ipsilateral femoral shortening, pure limb-length inequality, primary intrapelvic asymmetry, and decompensated scoliosis. The functionally low hemipelvis was lengthened as much as three centimeters by distraction at the osteotomy and insertion of a quadrangular bone graft. The option of a variable amount of acetabular redirection was especially useful in patients with ipsilateral acetabular dysplasia. Review of the patients after two to six years revealed that an average lengthening of 2.3 centimeters had been achieved. Fourteen patients had a balanced stance without a lift postoperatively, and the size of the lift required in the other six patients was greatly reduced.


Methylmethacrylate is a mutagen for Salmonella typhimurium. The long-term effects of exposure to monomeric methylmethacrylate have yet to be established. We have measured the toxicity and mutagenicity of methylmethacrylate for Salmonella typhimurium. At levels of thirty-four millimolar, methylmethacrylate exhibited 28 per cent of the mutagenic activity of an equimolar dose of dimethylnitrosamine. Methylmethacrylate alone exhibited toxicity to the bacteria, but when the methylmethacrylate was incubated with a rat-liver enzyme metabolizing system, mutagenesis was induced. These findings suggest that an intermediate metabolite of methylmethacrylate is mutagenic to Salmonella typhimurium.


Annual oration. Detection of abdominal abscesses: a combined approach employing ultrasonography, computed tomography and gallium-67 scanning. The identification and localization of intraabdominal abscesses is both an important and challenging task for the radiologist. Fortunately, three non-invasive imaging techniques are available which when used appropriately can virtually assure detection of an abscess. Consideration of the advantages and disadvantages of each modality will often indicate the most propitious sequence of investigations in the individual patients.


The spectrum of liver disease on CT.

The spectrum of liver disease on CT. Computed tomography (CT) is a useful imaging modality in a variety of liver diseases. The roles of radionuclide liver scanning, ultrasound and CT in liver disease are discussed and characteristic CT findings of cystic disease, trauma, abscess, primary and metastatic malignancy, fatty infiltration, hemochromatosis, and obstructive jaundice are illustrated.


Diagnostic imaging in diabetes mellitus. Diagnostic imaging techniques have an important role in supplementing the clinical evaluation of the diabetic. These are discussed in relation to the pathophysiologic changes: vascular degeneration, neurologic and infective changes, and maternal-fetal influences during pregnancy.


Quantitative reflection contrast microscopy of living cells. Mammalian cells in culture (BHK-21, PtK2, Friend, human flia, and glioma cells) have been observed by reflection contrast microscopy. Images of cells photographed at two different wavelengths (546 and 436 nm) or at two different angles of incidence allowed discrimination between reflected light and light that was both reflected and modulated by interference. Interference is involved when a change in reflected intensity (relative to glass/medium background reflected intensity) occurs on changing either the illumination wavelength or the reflection incidence angle. In cases where interference occurs, refractive indices can be determined at points where the optical path difference is known, by solving the given interference equation. Where cells are at least 50 nm distant from the glass substrate, intensities are also influenced by that distance as well as by the light's angle of incidence and wavelength. The reflected intensity at the glass/medium interface is used as a standard in calculating the refractive index of the cortical cytoplasm. Refractive indices were found to be higher (1.38--1.40) at points of focal contact, where stress fibers terminate, than in areas of close contact (1.354--1.368). In areas of the cortical cytoplasm, between focal contacts, not adherent to the glass substrate, refractive indices between 1.353 and 1.368 were found. This was thought to result from a microfilamentous network within the cortical cytoplasm. Intimate attachment of cells to their substrate is assumed to be characterized by a lack of an intermediate layer of culture medium.


Capping of exogenous Forssman glycolipid on cells. When motile cells are incubated with Forssman glycolipid, the antigen is incorporated into the cells' plasma membranes. If cross-linked by antibody, the patched glycolipids cap. This process is sensitive to those drugs that are known to inhibit capping of protein antigens. The results support a flow mechanism for capping.


Distribution of fetal bovine serum fibronectin and endogenous rat cell fibronectin in extracellular matrix. Normal rat kidney cells were cultured in medium supplemented with normal fetal bovine serum (FBS) or FBS depleted of fibronectin. The cell surface fibronectin of these cultures was visualized by indirect immunofluorescence using species-specific antisera for either rat fibronectin or bovine fibronectin. Anti-rat-fibronectin revealed fibrillar structures on the cells grown in either normal medium or fibronectin-depleted medium. Anti-bovine fibronectin revealed similar fibrillar networks, but only on the cells grown in medium containing bovine fibronectin. Staining in each case was abolished by absorption with the homologous antigen. It appears that exogenous fibronectin was incorporated into the same structures as endogenous fibronectin. This finding suggests that circulating fibronectin may serve as a building block for the assembly of extracellular matrix, possibly by cells which are incapable of synthesizing it.


Measurement of total, intracellular and surface bound cations in animal cells grown in culture. A procedure is described in which animal cells grown in culture on a dish are rapidly rinsed in situ with 0.25 M sucrose solutions for subsequent measurement of total, intracellular and rapidly exchangeab le Na+, K+, Mg2+ and Ca2+ by atomic absorption spectrophotometry. Repeated rinses with CO2-free (pH similar to 7) 0.25 M sucrose solution produced essentially no loss of cellular protein or cations. One 10-second rinse with CO2-saturated (pH 4) 0.25 M sucrose solution removed a rapidly proton exchangeable cellular cation fraction which is interpreted as being externally (membrane) bound. Rinses with physiological electrolyte solutions are shown to produce loss of cellular protein as well as displacement of surface exchangeable cations. Thus, isotonic sucrose solution is more satisfactory than electrolytic media for rinsing cultured cells prior to measurement of cellular cations. The technique employing sucrose rinse media is very rapid and reproducible and permits measurement of total, intracellular or surface bound Na+, K+, Mg2+ and Ca2+ in the same sample.


Prostacyclin stimulates the renin angiotensin aldosterone system in man. The effect of prostacyclin (PGI2) on the renin-angiotensin-aldosterone system (RAA) has been examined in 6 normal male volunteers infused with 3 incremental doses of PGI2. Diastolic blood pressure fell significantly during the infusion of the highest dose of 8 ng/kg/min. Plasma renin activity (PRA) and plasma angiotensin II (AII) increased to 271% and 342% respectively from baseline values. Plasma cortisol (PC), plasma noradrenaline (PNA) and plasma adrenaline (PAD) did not change significantly. Plasma aldosterone (PA) increased slightly 15 minutes after stopping the infusion. These results suggest aht PGI2 stimulates the release of renin in man.


Reliable urease test for identification of mycobacteria. A simple, modified formation for urea broth gave consistent, reliable results to aid in the differentiation of Mycobacterium species. In a study of 1,346 isolates representing 17 different species, tests read after 7 days were distinct and reproducible. The use of this test facilitates the identification of Mycobacterium species, such as M. scrofulaceum, the M. avium complex, the M. terrae complex, and M. triviale.


Merthiolate treatment of pathogenic fungi. The action of Merthiolate on the pathogenic yeasts Blastomyces, dermatitidis, Histoplasma capsulatum, and Sporothrix schenckii was compared to the effect of treatment with formaldehyde. Concentrations of 1:10,000 and 1:5,000 Merthiolate for three exposure times (24, 48, and 72 h) at 4 and 25 degrees C were tested on three media (brain heart infusion with and without blood, and modified Sabouraud agar). The effect of Merthiolate on these three yeasts was primarily fungistatic, with maximum effect using 1:5,000 Merthiolate at 25 degrees C for at least 48 h. Mycelial suspensions of B. dermatitidis, H. capsulatum, S. shenckii, and the yeast phase of Cryptococcus neoformans were susceptible to the 1:5,000 Merthiolate concentration after 24 h of treatment. The antifungal effect of Methiolate varies with species and growth phase of the fungus. Concentration, time of exposure, and temperature of incubation are important variables.


Nonfermentative bacilli: evaluation of three systems for identification. Three systems for the identification of nonfermentative bacilli were evaluated for their rapidity and accuracy of identification of 217 strains. Two of the systems, API 20E (API) and Oxi/Ferm tube (OxiF), are available as kits; the oxidative attack (OA) system is not commerically available. The overall accuracies of the OA, API, and OxiF systems were 91, 69, and 50%, respectively. Identification within 48 h was achieved for 98% of the strains by OA, for 50% by API, and for 18% by OxiF. Most of the organisms that were either misidentified or not identified by API and OxiF were those nonfermentative bacilli which are relatively more fastidious or rarely encountered or both. All three systems accurately identified nonfermentative bacilli commonly isolated at Olive View Medical Center, namely, Pseudomonas aeruginosa, Acinetobacter anitratus, Pseudomonas maltophilia, Acinetobacter lwoffi, saccharolytic flavobacteria (CDC IIb), moraxellae, Pseudomonas fluorescens, and Pseudomonas putida. The OA system identified 100% of the above organisms correctly, API identified 99.4%, and OxiF identified 99.3%. Since these organisms comprise 92% of the total number of nonfermentative bacilli isolated at Olive View Medical Center, we conclude that both API and OxiF may be useful alternatives to conventional methods, based on accuracy of identification alone. These two systems were considered substantially inferior to the OA system when both accuracy and rapidity of identification were taken into account.


Clinical evaluation of the MICRO-ID, API 20E, and conventional media systems for identification of Enterobacteriacea. MICRO-ID (General Diagnostics, Morris Plains, N.J.) is a new kit system designed for the identification of Enterobacteriaceae in 4 h. It consists of 15 biochemical tests of paper disks. Each test is in its own compartment in a molded plastic tray. Only one reagent need be added to the system (2 drops of 20% KOH, which is added to the Voges-Proskauer test). Based on the pattern of positive and negative biochemical test results, a five-digit octal code number is calculated. An identification is derived from a computer-generated identification manual. A study was conducted to compare three systems-the MICRO-ID 4-h and the API 20E (Analytab Products Inc., Plainview, N.Y.) 18- to 24-h systems and a conventional media system-to measure the ability of each to identify members of the family Enterobacteriaceae. Comparison tables, rather than simple percentage agreement tables, were generated to define the particular strengths and weaknesses of each system and allow the laboratory to best use the data. The MICRO-ID compared quite favorably with conventional media. MICRO-ID yielded incorrect identifications with 1.5% of the isolates tested (API 20E, 4.7% misidentification rate). Half the MICRO-ID misidentifications occurred when the system identified a Citrobacter diversus as a lysine-negative Escherichia coli; all gave one octal number. A direct comparison of the MICRO-ID and API 20E was of limited value because percentage agreements were merely the sums of the errors of each. The ease of inoculation, the requirement for the addition of only one reagent, and the 4-h capability make the MICRO-ID system an extremely attractive development in the field of bacterial identification.


Comparison of the API 20E and Corning N/F systems for identification of nonfermentative gram-negative rods. A total of 231 strains of nonfermenting gram-negative rods were tested on the API 20E system, the Corning N/F system, and conventional media. When the results of identification to species were compared, the API system agreed with the conventional system on 69% of the isolates, and the Corning system agreed with the conventional system on 79% of the isolates. Both kit systems were deficient in identifying Pseudomonas cepacia and the more unusual isolates.


Comparison of various McCoy cell treatment procedures used for detection of Chlamydia trachomatis. McCoy cells treated in six different ways, in addition to untreated cells, were compared to determine which gave rise to the largest number of Chlamydia trachomatis inclusions when tested with a laboratory-passaged strain. The same batch of cells was treated by irradiation, preinoculation exposure to cytochalasin B or 5-iodo-2'-deoxyuridine, and postinoculation exposure to cycloheximide, hydrocortisone, or emetine. Significantly more inclusions were always found in cells which had been treated with cycloheximide than in cells treated in any other way. Conversely, untreated McCoy cells always had significantly fewer inclusions than cells which had received some form of treatment. Similar results were obtained when cycloheximide-treated, irradiated, and untreated cells were inoculated with urethral specimens containing unpassaged organisms.