Epidemiological features of dengue and chikungunya infections in Burma. A serological survey for antibody to dengue and chikungunya was carried out in all 14 divisions and states and 2 border towns in Burma during 1973-74. Dengue HI antibody prevalence rate of less than 10% was observed in Arakan and Shan States, 10 to 30% in the Irrawaddy, Pegu, Mandalay Divisions and Kachin, Mon and Karen States, 31 to 60% in Sagaing Division, and over 60% in Rangoon, Magwe and Tenasserim Divisions. Similarly, chikungunya HI antibody prevalence rate of less than 10% was observed in Arakan State, 10 to 30% in the Irrawaddy, Pegu, Mandalay and Sagaing Divisions and Kachin State, 31 to 60% in Rangoon Division and Mon State. Both dengue and chikungunya antibodies were detected where Aedes aegypti mosquitoes were prevalent but the antibody prevalent rates were not directly proportional to the premises index. No HI antibody to dengue nor chikungunya was detected in Aedes aegypti free hilly areas, Chin and Kayah States, but was detected in the Shan State, Dengue and chikungunya infections were observed both in rural and urban populations. Dengue and chikungunya infections affected all socioeconomic classes in Rangoon equally but in Mandalay high socioeconomic class was nearly 3 times less affected than lower socioeconomic class. The infrequencies of dengue and chikungunya infections were observed to be 2 to 3 times higher in residents of Rangoon City than those of other towns. In Rangoon the antibody prevalence rates to dengue increased progressively with age while in other towns no appreciable increase in rates with age was observed. Both sexes were equally affected. This study provides strong circumstantial evidence that dengue and chikungunya viruses are highly and widely distributed throughout Burma, and that new outbreaks of haemorrhagic fever could occur in previously free areas following introduction of dengue viruses into populations previously exposed to one type of dengue.


The effects of free fatty acid infusion on glucose metabolism in normal and streptozotocin-treated dogs. By means of a novel technique for direct intravenous infusion of free fatty acids (FFA) into conscious dogs, it was found that the elevation of FFA levels in normal animals leads to a marked increase in insulin release and to a decrease in plasma glucose concentration. Dogs with streptozotocin-induced diabetes were used for FFA infusion in order to test whether peripheral glucose utilisation would be suppressed by FFA in the absence of the usual insulin response. Diabetic dogs receiving a constant infusion of intravenous glucose were given infusions of oleic acid after control periods of 2-3 hours. Constant intravenous infusion of U-14C-glucose into the diabetic dogs indicated that FFA infusion caused an average transient drop of 36,8% in glucose flux. After 1 1/2 hours of FFA infusion the flux returned to the former rates. Because no change in plasma glucose levels was found during FFA infusion our results can be explained only by a drop in hepatic glucose production concomitant with an equal drop in peripheral glucose utilisation.


Stimulation of the posterior columns of the spinal cord for relief of intractable pain. Sixty-nine patients with chronically incapacitating pain were treated with an implantable stimulator over the posterior columns of the spinal cord. Evaluations at 24, 30 and 34 months showed a progressive decrease in the number of patients considered to have an excellent result. Evaluation of 60 patients with pain of benign origin after implantation of the stimulating device showed only ten patients who could be considered to have an excellent result on the basis of their own report of pain relief. The most common failure of the stimulating devices was failure of stimulation into a painful part.


Electrical stimulation of the ventral and dorsal columns of the spinal cord for relief of chronic intractable pain: preliminary report. Serious questions are raised as to whether electrical control of pain is related to stimulation of the dorsal columns per se or the conduction of the electrical impulse via the cerebrospinal fluid to the ventrolateral columns. Percutaneous on-surface spinal cord stimulation is advocated as an accurate screening method before implantation. Of 27 patients who underwent dorsal column stimulator implants, the last three patients also underwent ventral column stimulator implants. Results in the patients with both implants tend to predict much better results with the possibility that ventral column stimulation alone is sufficient.


Abdominal wall replacement with plastic mesh in ablative cancer surgery. Planned excisions of tumors involving the abdominal or chest wall should not be compromised because of lack of tissue for wound closure. Reconstruction with an inert polypropylene mesh provides a strong and stable covering for the defect. Mobilization of adjacent skin flaps to provide covering for the mesh is desirable and results in reduced morbidity. When this is not possible the mesh may be left to granulate and later be grafted. Nine illustrative cases are presented.


The effects of small-bowel resection or bypass on the rat skeleton. The bones of adult rats became progressively osteopenic 1 to 5 weeks following jejunoileal bypass or resection. These changes were accompanied by increased levels of metaphyseal enzyme activities as well as by loss of histochemically identifiable osteoid. Osteoid tissue and the ability to mineralize skeletal collagen were recovered more rapidly and fully in the resection group than in the bypass group. Metaphyseal alkaline phosphatase concentrations increased in both groups coincident with the elevated lysosomal enzymes levels, and the skeletons showed a calcium deficit (low Ca/HOPr ratio) within the first 3 weeks. In resected rats the osteopenia and bone blood chemistry were consistent with hyperparathyroidism secondary to impared Ca absorption. In bypassed rats the results suggest that the osteopenia might be related to the release of a "resorptive factor" from the excluded intestinal segment.


A pulmonary hypertension-producing plant from Tanzania. An African youth who had died from primary pulmonary hypertension was suspected of having ingested a herbal remedy containing the seeds of the local plant Crotalaria laburnoides. Consequently powdered seeds of this plant were fed to 20 Wistar albino rats for 60 dyas to see if this would induce ventricular hypertrophy and associated hypertensive pulmonary vascular disease. At the end of the experimental period right ventricular hypertrophy, medial hypertrophy of the pulmonary trunk and 'muscular pulmonary arteries', and muscularization of the pulmonary arterioles had developed in a proportion of the test animals. These are the morbid anatomical features pathognomonic of a raised pulmonary arterial pressure and show that the seeds of Crotalaria laburnoides contain an agent capable of inducing pulmonary hypertension in rats. This study suggests the value of seeking a history of ingestion of herbal remedies and drugs in cases of unexplained pulmonary hypertension in man.


The in vitro synthesis of 7-hydroxy dehydroepiandrosterone by human mammary tissues. Normal and tumorous human mammary tissues were incubated in vitro with [7alpha-3H] dehydroepiandrosterone and [7alpha-3H] dehydroepiandrosterone sulphate. Tritium-labelled 7alpha-hydroxy dehydroepiandrosterone was identified as a principal metabolite from four of the five studies with normal tissue and all seven studies with tumorous tissue.


Biotransformation of pregnenolone - 7alpha-3H, progesterone - 7alpha-3H and dehydroepiandrosterone - 7alpha-3H by porcine fetal and maternal adrenal homogenate preparations. The biotransformation of pregnenolone-7alpha-3H and of progesterone-7alpha-3H by porcine fetal and maternal adrenal homogenates at 56 and 112 days of pregnancy and of dehydroepiandrosterone-7alpha-3H by fetal adrenal homogenates has been investigated in vitro. Both pregnenolone-7alpha-3H and progesterone-7alpha-3H were metabolized extensively by maternal adrenal preparations, the principal radioactive metabolites isolated being cortisol, corticosterone, 11-deoxycortisol, deoxycorticosterone, 11beta-hydroxyprogesterone and androstenedione. In addition, 17alpha-hydroxyprogesterone, 20alpha-dihydroprogesterone and cortisone were formed from both substrates and 17alpha-hydroxypregnenolone and progesterone were formed from pregnenolone. Although essentially the same radioactive metabolites were isolated after incubation of fetal adrenal glands with pregnenolone-7alpha-3H or progesterone-7alpha-3H, a greater proportion of the radioactivity was associated with corticosteroids at 112 days of pregnancy than at 56 days. 11beta-Hydroxyandrostenedione and androstenedione were isolated and identified together with an unknown polar metabolite, after incubation of fetal adrenal tissue with dehydroepiandrosterone-7alpha-3H. These results are discussed in relation to feto-placental steroid biosynthesis and metabolism and the role of the fetal adrenal in the initiation of parturition in the pig.


[Medical-dental specialty regulations and the educational progrm for "Specialist in Pedodontics"]. The Central Professional Committee for Paedodontics provides the instructors and the dentists specializing in paedodontics with an interpretation of the regulations for the diploma of specialist physician/specialist dentist of May 23, 1974, and of the educational program. Advice and recommendations are given for the fulfilment of the training program and also for the course and methods of the training for a dental speciality.


New mixed lymphocyte microculture test harvested by a simple device developed for this purpose. The methodology of the conventional mixed lymphocyte culture test has been simplified and miniaturized by developing a micromethod employing Terasaki trays with a final culture volume of 20 mul containing between 10,000 and 20,000 responding and stimulating cells each/culture. This represents one-tenth the cell concentration of the conventional mixed lymphocyte culture. For this new test, a special microharvester has been designed and developed. The test allows accurate and reproducible results giving a high level of discrimination between the different levels of response.


Blocking versus cytotoxic antibody in HL-A- and mixed lymphocyte culture-identical and nonidentical human renal transplant recipients. Blocking antibody directed against donor responding cells has been found to develop in HL-A-mixed lymphocyte culture-identical donor-recipient combinations after renal transplantation in which splenectomy was not performed. Cytotoxic antibody associated with rejection episodes and detected only by the discriminatory mixed lymphocyte culture developed in other HL-A-mixed lymphocyte culture-identical combinations as well. It disappeared as blocking antibody appeared and the post-transplant course became uneventful. In addition, both cytotoxic and blocking activity were shown to develop and coexist in a patient who received two successive renal transplants: cytotoxic against donor 1 (kidney rejected), and blocking against donor 2 (kidney accepted). These findings are taken to be strong evidence in favor of qualitatively demonstrable separate antibody molecules dealing with two separate functions, i.e., cytotoxicity versus enhancement or immunoregulation.


An evaluation of the mixed lymphocyte culture reaction in marmosets. Marmosets are unique in that all members may be considered to be natural blood chimeras because of the high frequency of fraternal twinning and placental vascular anastomoses between the fetuses. The mixed lymphocyte culture (MLC) reaction utilizing blood lymphocytes was evaluated to determine whether this in vitro test could detect histocompatibility differences among related and unrelated marmosets. It was found that the responses could be correlated with the probable immunogenetic relationships of these animals. Thus, an allogeneic MLC reaction in which the responding (R) and stimulating (S) cells were obtained from unrelated animals within one subspecies of marmosets (Saquinus fuscicollis illigeri) yielded a lower response than a semi-xenogeneic reaction involving R and S cells from two different subspecies of marmosets (S.f. illigeri (R) versus S.f. lagonotus (S). In contrast, MLC reactions between cells from chimeric marmoset co-twins were essentially negative, indicating specific immune tolerance. Under certain experimental conditions, however, it was suggested that the dual lymphocyte cell population from a chimeric marmoset could undergo "autostimulation" as measured by label incorporation studies. The necessity of further experiments was recognized before the data could be truly accepted as reflecting an in vitro manifestation of incompatibility between the two different genetic populations of blood lymphocytes from a chimera.


Glycogen synthesis by two Ehrlich ascites tumour cell strains in vitro. Cells of two closely related strains of the Ehrlich mouse ascites tumour synthesize glycogen in vitro. The glycogen is mainly deposited intranuclearly.


Light and electron microscopic observations on the carotid bodies of rats following adaptation to high altitude. The carotid bodies of rats were investigated by light and electron microscopy following adaptation to a simulated altitude of 7000 m. Some animals were studied immediately after readaptation to sea-level, other groups up to 41 days later. The animals of the first group show enlarged glomera with dilated capillaries as reported earlier. The type-I-cells are large with a light staining cytoplasm. The lobular configuration of the glomus is lost and intracapillary platelet thrombosis are frequently found. Under these thrombi there is usually a chief cell degeneration with edema and vacuolisation. There is also a noticeable decrease in so-called catecholamine bodies and the remaining ones are usually arranged along the cell borders. Electron microscopically the intercapillary tissue reveals an increase in collagen material partially replanning lost chief cells. Essentially unaltered sustentacular cells frequently enclose with their processes only collagen bundles and nerve fibres. During 41 days following the readaptation there is a narrowing of capillaries and vacuolisation, swelling and thromboses are no longer observed. The amount of collagen, however, remains the same. Dense cored vesicles again increase in number and the chief cells show a prominent interdigitation. Light microscopically the restoration to normal appears to be almost complete. Fine structural analysis, however, shows an appearently permanent distortion in the composition and cellular arrangement of the glomus.


Tissue specificity of the epidermal chalones. An increasing body of evidence indicates that the rate of cell renewal in labile and stable cell populations is regulated according to a negative feedback principle. Water extracts of such tissues contain mitosis-inhibiting substances (chalones) that probably represent the chemical signals in these cell populations. Tissue specificity is one of the theoretical prereguisites of the chalones but has not been proven in many cases. The present study is an attempt to evaluate the tissue specificity of some chalones, with special emphasis on the chalones found in keratinizing epithelium of the forestomach, and the epithelium of the crypts of jejunum and colon were chosen since they represent stady state systems with a well defined proliferating ppol...


The interaction between Walker tumour cells and mucosa cells in the lamina propria of gastric mucosa in rats. The tumour behaviour in previously X-irradiated mucosa compared to normal mucosa. One series of 12 rats was exposed to X-irradiation (1500 R) of the stomach 19 days before implantation of Walker tumour cells in the gastric mucosa, and the frequency of tumour take and the extent of tumour growth after 10 days were compared with a second series with the same tumour implantation, but without X-ray exposure. In a third series simple gastric ulcers without tumour were produced by clamping the gastric wall with a heated (80 degrees) surgical needle holder, and the animals were killed 5-7 day later. All the rats were given injections of vinblastine sulfate 3 hours and of 3H-TDR 1 hour before sacrifice. In viewfields with diameter 180 mu the vinblastine-arrested mitoses and labelled cells on the tumour side of the tumour/mucosa border were calculated as percentages of all tumour cells. In the mucosa the total number of proliferating cells was counted at various distances from the border of the tumour or ulcer. No clear differences in the frequency of tumour take and the extent of tumour growth were found between the X-irradiated and the normal rat stomachs, and it is concluded that the X-ray exposure 3 weeks prior to tumour implantation did not reduce the normal mucosal resistance to tumour growth. The percentage of arrested mitoses and labelled cells in the tumour decreased one view field away from the mucosal border, and the number of proliferating cells in the mucosa bordering on the tumours showed a gradual fall with increasing distance up to 0.8-1.0 mm from the tumour border; within these distances, however, the numbers were much higher than at corresponding distances from edges of the ulcers. The Walker tumour thus seems to stimulate cell proliferation in mucosa to a much greater extent than a simple ulcer does. The causes of this phenomenon and the possible roles of "chalones" or "anti-chalones" are discussed.


Generalized gangliosidosis type II (juvenile GM1 gangliosidosis). A pathological, histochemical and ultrastructural study. Pathological, histochemical and ultrastructural studies on 3 siblings with GM1 gangliosidosis type II are reported. These studies support a biochemical defect with profound deficiency of beta-galactosidases which results in widespread accumulation of the GM1 ganglioside and its asialo derivative in brain and to a lesser extent in viscera, as well as in storage of a keratan sulphate-like mucopolysaccharide. Striking valvular changes in the heart without myocardial involvement were seen in all cases. The histochemical and ultrastructural changes are similar to those seen in GM1 gangliosidosis type I, though less severe. Autosomal recessive inheritance without apparent ethnic predilection seems likely.


[Phagocytosis of group A haemolytic streptococci by mouse macrophages. Comparison of preliminary infection of animals and degree of virulence of the strain (author's transl)]. With the aim of studying immunological reactions associated with experimental streptococcal diseases, we have developed an in vitro technique for following phagocytosis of streptococci by peritoneal macrophages of mice. By this means we have been able to show that the rate of phagocytosis of streptococci by "normal" macrophages is inversely correlated to the degree of virulence of the strain. On the other hand, the rate of phagocytosis of the virulent strain is increased 10 times when the macrophages come from mice having previously been injected with this virulent strain.


Serum IgD and IgE in rheumatoid arthritis. Serum immunoglobulins IgD and IgE have been determined by a single radial immunodiffusion technique and a radioimmunoassay method in serum samples from 95 rheumatoid patients, 5 subjects with Sjögren's syndrome and 50 healthy controls, and compared with levels of IgG, IgM and IgA fractions measured in the same subjects. The IgD and IgE serum content resulted similar in the rheumatoid, Sjögren's and control sera. No correlation of IgD and IgE values with changes of other immunoglobulins or with the activity and the duration of the rheumatoid disease was observed.


[Calculation of the plausibility of paternity, using the results of blood grouping tests in cases in which the fatherhood of a man to twins and siblings is disputed. III. Practical experience with the method used for calculation of some cases concerning twins and siblings (author's transl)]. In 129 two-children cases, including 84 twin cases and 45 brother and sister cases, the probability of paternity of one or several men was calculated, applying the method of Hummel and Schulte-Mönting. With this method the whole information, given by the blood group results, is used. Because of this, the achieved values of probability in most of the cases are higher than the values achieved using the Essen-Möller formula, in which the relation between the children is not taken into account.


Virus infections in renal transplant recipients. 534 serum samples from 73 renal transplant recipients, 41 haemodialysis patients, and 99 blood and organ donors were examined serologically for antibodies against Cytomegalo, H. simplex (types 1 and 2), Varicella-zoster, Epstein-Barr, Adeno, Influenza, Parainfluenza, Respiratory syncytial, Measles, Picorna- and human Polyoma- Viruses. Serum specimens were stored in the lyophilized state until examined thus enabling a simultaneous testing of all samples belonging to one patient. All antigens, complement, and control antisera were prepared, lyophilized, and standardized in this laboratory. This has enabled the use of single batches of any preparation throughout the study. Serologic results with antigens of the Herpesvirus group (CMV, HSV and VZV) compared favourably with previous results showing that infections with these agents, especially with CMV, can frequently be encountered among transplant recipients. Our results have indicated a moderately increased incidence of infections with some Herpesviruses in haemodialysis patients as well. Infections with VZV, for instance, were as frequently demonstrated in these patients, as in transplant patients although the former received no immunosuppressive therapy. Serologic results with non-Herpesvirus antigens indicated an increased incidence of infections with Polyomavirus, Myxoviruses (Influenza, Parainfluenza and RS) and Picornaviruses among transplant recipients. The incidence of acute infections with RS virus among adults was unusually high and there is no evidence so far to indicate such a high frequency of RS infections in any other group of adults. We were unable to demonstrate acute infections with non-Herpesviruses among haemodialysis patients, even though most of the patients were followed over a period of more than 2 years. Virus isolation attempts were performed with samples of urines and biopsy or autopsy samples. 23 out of 28 cytopathic agents recovered from urines, throat-swabs and/or from organs of transplant recipients were identified as CMV. Two HSV type 1, 1 HSV type 2, and 2 Coxsackie B type 3 viruses were also isolated. No viruses were isolated from a series of 31 kidneys randomly selected among autopsy cases.


Immunoglobulins under the influence of nonspecific factors II. The influence of work-stress on levels of immunoglobulins (IgG, IgA, IgM) of miners in uranium mines. In 2 groups of 22 miners each, exposed 5.4 and 10.8 years respectively to the environment of uranium mines, the serum levels of IgG, IgA, and IgM were measured before and after one workshift. In the group longer exposed the work-stress resulted in a significant decrease of all 3 classes of Ig. In the group exposed for a shorter time only the IgG decreased significantly. In the group longer exposed the initial levels of IgG were considerably lower in comparison with the group exposed for a shorter time.


Chemical structure of the peptidoglycan, its modifiability and relation to the biological activity. The peptidoglycan is a heteropolymer composed of polysaccharide chains which are cross-linked through short peptides. The polysaccharide moiety (glycan) is made up of beta-1,4 glycosidically linked N-acetylglucosamine and N-acylmuramic acid residues. The carboxyl group of muramic acid is usually substituted by a peptide which consists of alternating L- and D-amino acids. These peptide subunits are cross-linked between the diamino acid in position 3 and the C-terminal D-alanine in position 4 of an adjacent peptide subunit either in a direct way or via an interpeptide bridge (Group A). In some coryneform bacteria the cross-linkage extends from the alpha-carboxyl group of D-glutamic acid in position 2 to D-alanine of a neighbouring peptide subunit (Group B). In the latter case a diamino acid is always found in the interpeptide bridge. A chemical modification of the peptidoglycan may occur in some bacteria due to growth in a quite unbalanced medium. The influence of glycine-rich and glycine-deficient growth medium on the chemical structure of the peptidoglycan of S. aureus will be discussed. Inhibiting concentrations of penicillin, glycine or D-amino acids can also modify the peptidoglycan. Further modification can occur through different extraction procedures which are used to obtain a clean peptidoglycan free of non-peptidoglycan cell wall material. Little is known about the molecular basis of the biological activity. The chemical composition is at least important for the antigenic determinants. The lysozyme susceptibility and the size of the preparation may be other crucial points for the biological activity of the peptidoglycan.


Chemical structure of the cell wall of Mycobacterium tuberculosis var. bovis, strain BCG. BCG cell walls contain approximately 30% free lipids like other mycobacterial cell walls. The insoluble skeleton of the cell wall is made up of two covalently linked polymers, a peptidoglycan and an arabinogalactan mycolate, with which are associated non peptidoglycan amino acids and a glucan. We present data on two structural features: 1. The "non peptidoglycan" amino acids; they form two kinds of compounds: peptide chains which can be solubilized by proteolytic enzymes and a trypsin-chymotrypsin insensitive poly-alpha-L-glutamic acid. 2. Presence of meso-DAP-meso-DAP1) interpeptide linkages in the peptidoglycan: this new type represents at least 50% of the interpeptide linkages of the cell wall of the BCG strain.


Estimates of the porosity of Bacillus licheniformis and Bacillus subtilis cell walls. The maximum porosity of Bacillus subtilis and Bacillus licheniformis cell walls was estimated by two independent and relatively simple methods. Peptidoglycan was isolated from Bacillus subtilis cell wall preparations and used as an insoluble support for exclusion chromatography of dextrans of known average molecular size. In an alternative approach the leakage of radioactively labelled proteins from Bacillus licheniformis cells incubated in butanol-saline mixtures was measured and their size estimated by exclusion chromatography on Sephadex G-100. Under these conditions the permeability barrier of the cytoplasmic membrane was destroyed with preservation of the structural integrity of the outer cell wall. The apparent exclusion threshold of the cell wall of either organism as determined by these means corresponded to molecules with a diffusional radius of not more than 2.5 nm.


Immunochemical studies with synthetic immunogens chemically related to peptidoglycan. Rabbits were immunized with synthetic immunogens HSA-(Gly-L-Ala-L-Ala-D-Ala-D-Ala)39 and HSA-(Gly-gamma-D-Glu-L-Ala-D-Ala-D-Ala)40, respectively. Antibodies against HSA-(Gly-L-Ala-L-Ala-D-Ala-D-Ala)39 showed a strong precipitin reaction with the homologous antigen, with HSA-(Gly-gamma-D-Glu-L-Ala-D-Ala-D-Ala)40 and with solubilized peptidoglycan containing peptide subunits with C-terminal D-alanyl-D-alanine. The albumin-peptide conjugates also cross-reacted with rabbit antisera to Streptococcus A-variant, which contain antibodies directed against the peptide moiety of peptidoglycan. The proof for identical determinant groups of peptidoglycan of Streptococcus A-variant and HSA-(Gly-L-Ala-L-Ala-D-Ala-D-Ala)39 was furnished by Ouchterlony gel diffusion studies and by the appropriate inhibition tests of the quantitative precipitin reaction. Immunization of rabbits with HSA-(Gly-gamma-D-Glu-L-Ala-D-Ala-D-Ala)40 yielded antisera which, besides the specificity of antisera to HSA-(Gly-L-Ala-L-Ala-D-Ala-D-Ala)39, showed an additional specificity.


Estimation of peptidoglycan antibodies by an immunoperoxidase technique. An immunoperoxidase technique developed for the detection and measurement of anti-peptidoglycan antibodies is described. The technique is based on the Mancini single radial immunodiffusion test with the addition of a reaction step with peroxidase-labelled anti-immunoglobulins. The dark-brown colour produced by the enzyme and a H2O2-diaminobenzidine solution increased the sensitivity considerably. The test was found to be highly specific and valuable both for the screening of total amounts of anti-peptidoglycan antibodies in sera as well as in studies of antibody specificity.


Antibodies to peptidoglycan in the sera from population surveys. A recently described latex agglutination test was used to determine peptidoglycan antibody titers in sera from healthy human subjects and in sera from patients with a history of streptococcal infections or rheumatoid arthritis. Using latex particles coated with group A streptococcal peptidoglycan 32.8% of the sera from 961 healthy donors reacted positively with titers ranging from 1 : 5 to 1 : 320. Peptidoglycan antibodies were more frequently present in the younger population (45.1% in the 20-29 years old) and considerably decreased in advanced age (15.7% in the 70 years or older). Sera from 82 patients with elevated ASO-titers showed detectable peptidoglycan antibody levels in 40.2%; a statistically significant correlation between ASO and peptidoglycan antibody titers could not be substantiated. Sera from 25 patients with rheumatoid arthritis gave a high incidence (56-92%) of positive results. However there was evidence that this may be due to the action of rheumatoid factor present in such sera.


Rheumatoid factor appearance in Micrococcus lysodeikticus immunization and its interference with allotype specific reactions. Rabbits hyperimmunized with Micrococcus lysodeikticus produce, in addition to anti-cell wall antibodies, considerable amounts of anti-immunoglobulins or rheumatoid factors which can interfere with immune reactions. In particular, in immunodiffusion, they can reveal non precipitating systems for which the immunodiffusion typing sera are usually not tested. In our hands, rabbit allotypes d12 and A8 give rise to visible immunodiffusion reactions in presence of rheumatoid factors. Inhibition of this reaction can be used to type for these markers in sera which do not contain the rheumatoid factors.


Chemical structure of lipopolysaccharides and endotoxin immunity. Lipopolysaccharides (endotoxins) of gram-negative bacteria consist of 2 components with distinct physico-chemical character: a heteropolysaccharide and a covalently linked lipid, termed lipid A. The chemical structure of lipid A, which represents the toxic center of lipopolysaccharides, is discussed. Evidence is presented that lipid A antiserum suppresses the pyrogenic effect of lipid A and lipopolysaccharides in rabbits. The protective power of lipid A antiserum, however, is only expressed in animals which have been pretreated with lipid A or lipopolysaccharide indicating that other than humoral factors, perhaps cellular, also participate in endotoxin fever (cross) immunity. The fever resistance mediated by lipid A antiserum seems to be endotoxin-specific with regard to both the preparative and the challenging injection. Lipid A antiserum therefore may serve as a tool to discriminate between fever caused by endotoxins and that induced by other pyrogens.


Physical state and biological activity of lipopolysaccharides. Toxicity and immunogenicity of the lipid A component. Bacterial lipopolysaccharides are negatively charged macromolecules due to the presence of phosphate, pyrophosphate and carboxyl groups. When isolated from bacteria, they are obtained in salt form with metal cations and basic amines. Removal of these ionically bound substances by electrodialysis leads to acidic lipopolysaccharides which on neutralizing with different bases, preparations are obtained which show distinct differences in their physico-chemical properties and in their biological activity. Soluble lipopolysaccharides interact with complement leading to loss of hemolytic activity. This property is embedded in the lipid A part of the molecule and is expressed only when the lipopolysaccharide is present in a favourable particle size. Nevertheless, a number of lipopolysaccharides exists, which, regardless of their particle size do not interact with complement. Lipid A is the part of the molecule responsible for endotoxicity. This was demonstrated by employing solubilized lipid A in complex form with BSA. Soluble lipid A/BSA complexes proved highly toxic for mice and pyrogenic in rabbits, and express many biological activities exhibited by intact lipopolysaccharides. Lipid A, when exposed on the bacterial cell-surface acts as a powerful immunogen, giving rise to the production of specific anti-lipid A antibodies that interact with the lipid A obtained from lipopolysaccharides that are otherwise distinct in their O-serological specificity. Anti-lipid A antibodies occur naturally in the serum of many animals and humans. The biological significance of anti-lipid A antibodies is discussed.


Endotoxin-like properties of the peptidoglycan. Peptidoglycan is responsible for the endotoxin-like properties of the streptococcus cell wall. The pyrogenic response of rabbit to group A streptococcus peptidoglycan prepared by hot formamide or TCA is dose-dependent and is increased if the material is ultrasonically solubilized. The pyrogenicity can be eliminated by the antiserum to the peptidoglycan or by the degradation of the material by lysozyme. Peptidoglycans prepared from cell walls of group B and L streptococci, Staphylococcus aureus, Staphylococcus epidermidis and Streptococcus pneumoniae produce fever effects comparable to the response after group A streptococcus peptidoglycan. Spirillum serpens and Escherichia coli contain in addition to endotoxin the peptidoglycan which is also pyrogenic. Repeated injections of bacterial peptidoglycan to rabbit result in tolerance to the fever effect. Cross-tolerance was recorded only exceptionally. Rabbits tolerant to endotoxin respond with a lower fever to S. aureus and group A streptococcus peptidoglycans. Intravenous administration of peptidoglycan to rabbit causes extensive alterations in the heart characterized by various stages of the degenerative and necrotic process. Local Shwartzman reaction can be elicited in rabbit by peptidoglycan used either as a preparative or as a provocative dose in combination with endotoxin, or it can be used for both doses. The results obtained with peptidoglycans prepared from various bacteria are fully comparable. Non-specific resistance of mice to infection induced by streptococcus cell walls was found to be dependent on the peptidoglycan activity; cell wall proteins and polysaccharide are inactive. These properties of peptidoglycan resemble those known from endotoxin studies. The data presented suggest the role of peptidoglycan in pathological reactions resulting from host-parasite interaction.


Biological properties of the peptidoglycan. This review deals with those biological activities of peptidoglycan that are not directly analogous to the properties of gram-negative bacterial endotoxin. The report is divided into 3 major parts: 1. A survey of peptidoglycan activities such as the induction of inflammatory skin reactions, lesion-enhancing activity (virulence factor), inhibition of phagocytosis of bacteria, inhibition of cell migration, cytotoxicity to mammalian cells, potentiation of the humoral and cellular immune response (adjuvant activity) and enhancement of tumor defense in experimental animals. 2. A presentation of factors which may influence these biological activities of peptidoglycan. 3. A brief discussion of the potential mechanisms of action of peptidoglycan.


Use of limulus assay to compare the biological activity of peptidoglycan and endotoxin. The gelation of a lysate prepared from the amebocytes of Limulus polyphemus, the horseshoe crab, has been used to detect endotoxin-like material in clinical studies and in "in vitro" experiments. The investigation of blood samples from 54 hospitalized patients suspected of having endotoxemia, revealed a positive limulus test in 14. Infections due to gram-positive organisms were not associated with positive assays. These results were in agreement with the observation that living gram-positive microorganisms - in contrast to gram-negative bacteria - did not initiate lysate gelation when studied "in vitro". Only very high concentrations of peptidoglycan, isolated from the cell walls of various gram-positive bacteria, induced the reaction. Therefore, these findings support the view, that the limulus test is relatively specific for the detection of gram-negative bacterial endotoxin and endotoxemia.


The thrombocytolytic activity of bacterial peptidoglycans. The peptidoglycan of group A Streptococcus prepared by hot formamide or TCA extraction has an expressive ability to lyse rabbit blood platelets in vitro. Initial changes in the ultrastructure of platelets can be observed after 10 minutes incubation of rabbit platelets with 0.1 mug of Streptococcus peptidoglycan per ml. The submicroscopic alterations are characterized by changes of the shape and damage of the limiting membrane of the platelets. Larger amounts of peptidoglycan (10 mug/ml and more) cause total destruction of platelets; only remainders of the limiting membrane and free granulomers can be seen. Peptidoglycans of Streptococcus pneumoniae and Staphylococcus aureus exhibit a comparable effect on rabbit platelets. There are substantial differences in the sensitivity of blood platelets of different animal species to the streptococcal peptidoglycan. Rat platelets exhibit a similar thrombocytolytic effect after the Streptococcus peptidoglycan treatment as rabbit ones; however, the development of comparable changes in their submicroscopic structure needs a dose of peptidoglycan 10(2)-10(3) times higher. Platelets of guinea-pig, dog, cat, calf and human appear to be quite resistant under analogous conditions to as such a high dose of Streptococcus peptidoglycan as 500 mug/ml.


The effect of some bacterial products on temperature and sleep in rat. The lipopolysaccharides from P. aeruginosa, S. minnesota and mucopeptide from Streptococcus group A injected intravenously into rats induce a dose-dependent changes of temperature. Simultaneously, a profound disturbance of sleep occurs. The administration of salicylate, which markedly suppressed the fever does not influence the mucopeptide-caused sleep disturbance. The most prominent change in the sleep pattern is a marked decrease of the total time of paradoxical sleep. The measurement of turnover rates of 5-hydroxytryptamine (5-HT) and noradrenaline (NA) in hypothalamus and midbrain, areas involved in temperature and sleep control, after injection of streptococcal mucopeptide demonstrated a significant increase of 5-HT turnover in both areas during fever and paradoxical sleep deprivation. Small electrolytic lesions of the dorsal raphe nuclei which are the largest collection of neural cells containing 5-HT completely eliminated the pyrogenic potency of mucopeptide. The findings suggest that some bacterial products might increase the body temperature through the interference with activity of 5-HT-containing neurons of the raphe complex.


Interaction of bacterial cell wall polymers and rat macrophages. Processing of group A and group D streptococcal cell wall was measured after phagocytosis by normal rat peritoneal cells in tissue culture. Group A cell wall was practically non-biodegradable in contrast to group D, which was over 80% degraded by 4-8 days in culture. There was no difference in elimination or degradation of mucopeptide or polysaccharide of group A cell walls. Neither antiserum or sensitized lymphocytes affected persistence. Macrophages from Fisher rats (susceptible to group A cell wall-induced polyarthritis) became cytotoxic for target L-cells 6-7 days after ingestion of group A cell walls. Phagocytosis of group D cell walls induced less cytotoxicity. Macrophages from Buffalo rats (resistant to polyarthritis) were less cytotoxic after phagocytosis of group A cell walls than Fisher macrophages. Soluble cytotoxins could not be detected in macrophage culture media.


Migration inhibition of peritoneal macrophages by peptidoglycan. Previous studies have shown that peptidoglycans from group A and B streptococci inhibit the migration of peritoneal exudate cells from non-sensitized rats and guinea pigs. In the present studies peptidoglycans from S. aureus and S. epidermidis were found to inhibit the migration of cells to the same extent as group A streptococcal peptidoglycan. In contrast, HSA-pentapeptide, an immunologically active synthetic analog of the peptide moiety of peptidoglycan which is free of the intrinsic toxicity of naturally occurring peptidoglycans, did not induce migration inhibition of peritoneal exudate cells from non-sensitized guinea pigs. Sensitization of animals with 200 mug HSA-pentapeptide emulsified in incomplete Freund's adjuvant significantly reduced the inhibitory effect of streptococcal and staphylococcal peptidoglycan; HSA-pentapeptide again showed no activity. However, when HSA-pentapeptide was tested against cells from animals sensitized with 200 mug HSA-pentapeptide incorporated in complete Freund's adjuvant, a strong inhibition of migration was evident. Skin tests performed in these animals, in contrast to the dermonecrotic reaction elicited by streptococcal or staphylococcal peptidoglycan, revealed a characteristic delay hypersensitivity to HSA-pentapeptide.


Cell-mediated immune reactions in vitro to cell walls and peptidoglycan from Staphylococcus aureus. Migration inhibition factor (MIF) was produced by peritoneal exudate (PE) cells from guinea pigs sensitized with heat-killed cells of Staphylococcus aureus cultured in vitro with staphylococcal cell walls or defined subunits of the cell walls. MIF activity was assayed by inhibition of migration of alveolar macrophages from lungs of normal guinea pigs. Specificity of inhibition was established using PE cells from tuberculin sensitive guinea pigs and tuberculin as appropriate controls. Staphylococcal cell walls, their peptidoglycan complex, and teichoic acid-peptidoglycan fragments stimulated MIF production by staphylococcus sensitive PE cells. Peptidoglycan fragments and teichoic acid were ineffective antigens.


Immunoadjuvant activities of cell walls, their water-soluble fractions and peptidoglycan subunits, prepared from various gram-positive bacteria, and of synthetic n-acetylmuramyl peptides. 1. The cell walls from some 20 species of gram-positive bacteria, with only few exceptions, were found to be definitely adjuvant-active in both stimulation of increased serum antibody levels and induction of delayed-type hypersensitivity to ovalbumin when administered to guinea pigs in the form of a water-in-oil emulsion. 2. By the use of various cell wall lytic enzymes, the immunoadjuvant principles were solubilized with the full retention of adjuvant activities observed with the cell walls of S. aureus, Str. pyogenes, Str. mutans, L. plantarum, C. diphtheriae, Myc. smegmatis and A. viscosus. N-acetylmuramyl peptide monomers (either L-Lys or meso-Dap type) were shown to be the unit chemical structure responsible for the manifestation of adjuvant activities to stimulate both antibody-mediated and cell-mediated immune responses. 3. Several N-Acetylmuramyl peptides were prepared by condensation of benzyl N-acetyl-4,6-O-benzylidene-alpha-muramide with each peptide benzyl ester by means of dicyclohexylcarbodiimide-N-hydroxysuccinimide method and removal of the protecting groups by hydrogenolysis. N-acetylmuramyl-L-alanyl-D-isoglutamine was identified as the minimum structural entity essential for the immunoadjuvant activities characteristic of bacterial cell walls. Neither synthetic N-acetylmuramyl-L-alanine nor L-alanyl-D-isoglutaminyl-L-lysyl-D-alanin was found to be active.


Interaction of peptidoglycans with anti-IgGs and with complement. This report describes the interaction of peptidoglycan (Streptococcus group A, Staphylococcus epidermidis and Micrococcus lysodeikticus) with 2 serum mediator systems, namely with the anti-IgG system and with complement. The observation that the majority of rabbits hyperimmunized with A-variant streptococcal vaccine produced anti-group carbohydrate antisera containing anti-IgGs and antibodies directed to peptidoglycan suggested that the production of these 2 latter antibodies was related. This view was supported by the finding of a monoclonal 7S anti-IgG with antibody specificity for the pentapeptide of peptidoglycan as evidenced by inhibition of the coprecipitation of 7S anti-IgG with antigen-antibody complexes by the pentapeptide. Inhibition of the anti-idiotype reaction by the pentapeptide provided further evidence for the antibody specificity of 7S anti-IgG for peptidoglycan. When added to normal human sera all peptidoglycan preparations inhibited the hemolytic activity of the sera. Consumption of C3 in C2 deficient serum and consumption of C2 in normal serum indicated the activation of both known complement pathways. Activation of the classical pathway of complement was more efficient since 50 mug of peptidoglycan consumed approximately 70% of C2 per ml normal serum whereas more than 2 mg of the same preparations was required to inactivate 17-24% of C3 in C2 deficient sera. Each of the different peptidoglycan preparations consumed similar amounts of complement in all 20 sera tested. This finding suggested that activation of the classical complement pathway by peptidoglycan was not mediated by anti-peptidoglycan antibodies present in only 20-40% of normal human sera.


Peptidoglycan adjuvants: minimal structure required for activity. The chemical structure of the adjuvant active fraction of mycobacterial cell walls has been investigated. It had been shown previously that soluble peptidoglycan fragments obtained from cell walls of Mycobacteria by lysozyme digestion or by other treatments act as adjuvants for increasing both humoral and cellular immunity. We then found that even the monomer subunit of the peptidoglycan of Mycobacteria (i.e. a disaccharide-tetrapeptide) is adjuvant active; then, similar compounds from other strains of bacteria were tested; the monomeric subunits of meso-diaminopimelic acid as well as L-lysine containing peptidoglycans were found to be adjuvant active. The smallest active compound studied so far is N-acetyl-muramyl-L-alanyl-D-isoglutamine synthesized for us by SINAY et al. (1975).


Adjuvant activity of bacterial peptidoglycans. Purified peptidoglycans isolated from various Gram negative and Gram positive bacteria can substitute for Mycobacteria in Freund's adjuvant. Monomeric subunits possess the same adjuvant activity. This activity seems to be related to the peptidic fraction.


[The preparation and characterization of transfer factor (author's transl)]. A transfer factor (TF) was prepared and characterized from the leucocytes of Mantoux-positiv donors using a variety of techniques. The results allowed the following conclusions: Whilst extraction by dialysis and phenol (without precipitation) led to nearly the same positiv immunological activities, when assayed in the MEM-test, phenol extracts subjected to RNA-precipitation lost considerable activity. The residual activity was associated with the lower molecular fractions, and RNA itself has shown no activity. Residues after leucocyte dialysation have also shown considerable TF-activity when extracted by phenol. It was found that phenol extraction followed by gel filtration allowed recovery of considerable quantities of active TF, confined to several specific filtration fractions. Combination of active and inactive gel filtration fractions suggested that there existed an inhibitory factor within the extracted leucocyte preparations. This modified preparation and purification procedure may allow the preparation of TF for therapeutic purposes.


Fractionation of antigen reactive cells from immunized mice on columns coated with antigen or anti-immunoglobulin sera. Immunocompetent cells obtained from NIP-RGG immunized mice were fractionated on bead columns coated with antigen or anti-immunoglobulin serum. The separated cell fractions were examined for their capacity to be stimulated by the antigen in short term culture, to produce antigen specific antibodies in the plaque assay and to bind radioactive labeled antigen. Cells which produce hapten specific antibodies or bind radioactive labeled hapten are removed from the cell population passed through a hapten-carrier complex coated column. Cells stimulated by the antigen to an increased DNA-synthesis are also retained by columns coated with the hapten-carrier-complex or the carrier alone; the fractionation seems to be carrier specific. The fractionation of cells is blocked by free antigen in the columnar fluid. However, the fractionation patterns of cells passed through anti-Ig-serum coated columns are different when antibody producing cells and cells stimulated by the antigen are compared. Whereas antibody producing cells and antigen binding cells are almost completely retained by anti-Ig-serum coated columns the cells which are stimulated by the hapten carrier complex are not removed from the passed cells. Studies to characterize the fractionated cell populations according to their sensitivity to anti-theta-serum, to the presence of Ig-receptors and to the phytohemagglutinin stimulation indicate that the antibody producing cells and the antigen binding cells have to be attributed to B-cells whereas the question whether the antigen stimulated cells are T- or B-cells cannot be definitely answered.


Interactions of the third component of complement (C3) with cross-linked dextran. III. Isolation and characterization of interacting components after enzymatic digestion of complement coated cross-linked dextran. In previous investigations we could show that incubation of cross-linked dextran (Sephadex) with normal human and normal guinea pig serum results in the binding of C3 to Sephadex. This binding was found to be due to activation of C3 via an alternate pathway. In this paper data are presented which show that components bound to Sephadex can be recovered after enzymatic digestion of serum-reacted Sephadex beads. The digests were characterized with immuno-electrophoresis and double immunodiffusion techniques. It could be shown that the main component present in the digest was converted C3. Apart from C3 under our test conditions only minute amounts of C3A but no other serum proteins were detectable. The observation that almost exclusively C3 is bound to Sephadex is further supported by the finding that immunization of rabbits with serum-reacted Sephadex beads results in the exclusive formation of anti C3 antibodies. Implications from these findings and possible applications are discussed.


Characterization of a stimulating factor released by immunosuppression. To get more information about the mechanism of stimulation of the immune response during immunosuppression under certain conditions, stimulating factor (SF) was collected 48 hours after treatment of NMRI-mice with cyclophosphamide. SF was characterized by different steps of separation using centrifugation, dialysis and salt-fractionation. For demonstration of the SF animals were sensitized with SRBC and given different substrates separately. The stimulation of the antibody-production was demonstrated on the cellular level using the plaque-technique. SF was detected by comparing the counts of plaque-forming cells in the spleen of treated mice compared with controls merely sensitized or given the same substrate of normal animals. In this way it was shown that the SF is a soluble factor in the serum with a molecular weight of more than 20,000, but that it is no immunoglobulin.


[A method to calculate the plausibility of paternity using blood group results of any relatives (author's transl)]. A calculation program is described by means of which the frequency of any type of relationship in cases of disputable descent (questionable partenity, maternity, parenthood, brother- and sisterhood) using blood group results, can be calculated. Within the scope of forensical practice, the program is above all important in cases in which the alleged father and/or mother of the child is deceased, where however, blood group results of other relatives are at disposal, and also in cases of suspected incest - when the alleged father of the child is at the same time father or brother of the child's mother, and, finally, in cases where two men who are related to one another are considered as possible fathers. The program can also be applied to search cases.


Cell-mediated immunity to cell wall antigens of Staphylococcus aureus in vitro. II. The production of chemotactic factor by guinea pig peritoneal exudate cells. Peritoneal exudate cells from sensitized guinea pigs were incubated in culture with staphylococcal cell walls. Filtered supernatant fluids from the cultures were chemotactic for horse leukocytes. Maximum chemotactic activity was observed in supernatant fluids of cultures incubated 1.5 hours. Activity disappeared after 4.5 and 18 hours additional incubation.


HL-A antigens in idiopathic Addison's disease. The frequencies of 36 HL-A antigens of the SD-1, SD-2 and SD-3 locus have been determined in 20 patients with idiopathic Addison's disease and compared with those of 450 controls. No significantly different HL-A antigen frequencies could be observed in both groups. These findings are in contradiction to data reported by a Copenhagen group. Possible mechanisms which might have led to this discrepancy are discussed.


Interactions of the third component of complement (C3) with cross-linked dextran. IV. Adherence of lymphocytes to C3 coated dextran gel particles. In previous studies we could show that the third component of complement (C3) is bound from normal serum to cross-linked dextran (Sephadex) after activation via an alternate pathway. Data presented in this paper demonstrate that a subpopulation of human lymphocytes adheries to C3 coated Sephadex particles. The adhering subpopulation of lymphocytes is identical with or overlaps extensively with lymphocytes bearing easily detectable membrane Ig. This phenomenon can be used for the fractionation of lymphocyte subpopulations. After a single passage of tonsillar lymphocytes through C3 coated Sephadex bead columns, no membrane Ig positive lymphocytes were detectable in the effluent. 97-100% of effluent cells formed rosettes with untreated sheep red blood cells, thus resembling pure T cells. Apart from that, the use of C3 coated dextran gel particles allows also the recovery of adherent cells after dextranase digestion of the dextran gel particles. An investigation of retained cells showed enrichment of B cells, but also T cells were demonstrable. The retention of these T cells is probably due to nonspecific interactions.


Formation and composition of the C3 activating enzyme complex of the properdin system. Sequential assembly of its components on solid-phase trypsin-agarose. An active C3 cleaving enzyme is generated when properdin factor B (glycine-rich beta-glycoprotein, GBG) is activated (GBG is cleaved) by factor D in the presence of C3b and Mg++. Factor D can be replaced by trypsin in this reaction but even then C3b and Mg++ are required. In the absence of C3b and/or Mg++ trypsin does not generate C3 cleaving activity from GBG although it cleaves GBG and releases its GGG fragment (B) under these conditions as well. Addition of C3b to GGG immediately after its release does not yield a C3 cleaving enzyme. These findings indicate that C3b, GBG and Mg++ interact, and that only in association with C3b can GBG be cleaved in a way that its enzyme activity, residing in a C3b, GGG complex, is expressed. The complex is labile (half-life at 20 degrees C: 9 minutes); Mg++ does not affect its stability nor is it essential for the activity. It was possible to sequentially fix on agarose the essential components of the C3 cleaving enzyme and thus to elucidate the single steps and the order of its formation. C3 was activated and the resulting C3b fragment fixed on agarose by incubating C3 with trypsin covalently bound to the agarose. The agarose-C3b intermediate was capable of binding GBG provided Mg++ was present. GBG could then be cleaved by factor D or trypsin added in solution; the solid-phase-fixed complex obtained had C3 cleaving activity, in the presence as well as absence of Mg++. Omission of any of these steps or components, or changes in the sequence did not give rise to an active enzyme. Mixtures of C3b, GBG and Mg++ have weak C3 cleaving activity by themselves. C3 cleavage in such incubation mixtures proceeds slowly for hours and is not accompanied by cleavage of GBG. There is thus complete analogy between the CVF-dependent and C3b-dependent C3 cleaving systems. C3b and CVF act in the same way, they form a reversible, weakly active C3 cleaving complex with GBG and Mg++, the activity of which is markedly enhanced but becomes subject to decay when GBG is cleaved by trypsin-like enzymes while bound to CVF or C3b.


[Blood group opinion and judicial judgement 1. 250 paternity cases 1965-1970 (author's transl)]. In 250 paternity actions (one-man one-child affairs 1965-1970) without exclusion by blood group opinion, which include 48 cases involving persons from abroad, 41 men acknowledged the paternity or withdrew the suit during the process. 32 cases were judged by default. 174 men were judged of child support according to the former 1717 BGB, 13 of these after an anthropological expertise. In 2 cases the anthropological opinion gave an indication of nonpaternity; accordingly, the court dismissed the action. In 157 cases the plausibility of paternity according to Essen-Möller's formula was calculated by the expert and dealt with in the blood group opinion. It appears, however, that the W values had no remarkable influence on the decision of the court.


Peripheral blood T and B lymphocytes in men in different age groups. Peripheral blood T and B lymphocytes were determined in normal humans at different ages. Spontaneous rosette formation with sheep red blood cells (SRBC) was used as a marker for T cells. B cells were detected by immunofluorescent staining of membrane-bound immunoglobulins. Blood samples from old individuals contained significantly lower T lymphocyte numbers than those from children. This diminution of circulating T cells caused a reduction of the total lymphocyte count in the elderly persons. No significant differences were between the T cell values of young and old adults. Whereas the percentages of B cells indicated an increase of this lymphocyte population in old humans, the absolute numbers of B lymphocytes remained almost unchanged during aging.


Chemical syntheses and properties of hydroxycinnamoyl-coenzyme A derivatives. Acyl-CoA derivatives of several hydroxylated cinnamic acids have been synthesized in 30 to 50% yield via a. acyl phenyl thiol esters, b. acyl N-hydroxysuccinimide esters, and c. glucocinnamoyl derivatives. Properties of the intermediates have been determined. The cinnamyol-CoA thiol esters were characterized by their chromatographic behaviour and UV spectra. The molar extinction coefficients of these important intermediates in plant phenylpropane metabolism have been unequivocally determined. Recently published values 13 for the molar extinction coefficients of these derivatives are incorrect; the methodological reason for this error has been established.


On the origin of protein biosynthesis.

On the origin of protein biosynthesis. There is a very close steric relationship between the codon-anticodon site which accounts for the genetic code dictionary and a polynucleotide replicase site. Protein biosynthesis must therefore have arisen out of a primaeval polynucleotide replicase system.


ATPase activity and light scattering of acto-heavy meromyosin: dependence on ATP concentration and on ionic strength. 1. The dependence on ATP concentration of ATPase activity and light scattering decrease of acto-HMM could be described at very low ionic strength by one hyperbolic adsorption isotherm with a dissociation constant of 3 X 10(-6)M. Hence the increase of ATP ase activity was paralleled by a decrease in light scattering. At higher values of ionic strength ATPase activity stopped rising before HMM was completely saturated with ATP. Higher ionic strength prevented ATPase activity from further increasing when the rigor links (links between actin and nucleotide-free myosin), which have formerly protected the ATPase against the suppressing action of higher ionic strength have fallen below a certain amount. This protecting influence of rigor links did not require tropomyosin-troponin. 2. For complete activation of ATPase activity by actin less actin was needed when HMM was incompletely saturated with ATP than when it was completely saturated with ATP. 3. The apparent affinity of ATP to regulated acto-HMM (which contained tropomyosin-troponin) was lower than to unregulated acto-HMM (which was devoid of tropomyosin-troponin). In the presence of rigor complexes (indicated by an incomplete decrease of light scattering) the ATPase activity of regulated acto-HMM was higher than that of unregulated acto-HMM. At increasing ATP concentrations the ATPase activity of regulated acto-HMM stopped rising at a similar degree of saturation with ATP as the ATPase activity of unregulated acto-HMM at the same ionic strength.


Inhibition of excision repair without influence upon UV-sensitivity and UV-mutability in Escherichia coli B/r Hcr+. Pre-irradiation starvation of exponentially growing Escherichia coli B/r Hcr+ thy-try- strain for thymine and tryptophan causes inhibition of pyrimidine dimer excision from ultraviolet damaged cells DNA. This inhibition of excision repair has not resulted in increasing ultraviolet sensitivity nor in increasing frequency of ultraviolet induced tryptophan revertants. The possible mechanisms of the non-excision repair in the prestarved cells, which is at least as accurate and effective as the whole dark repair in exponentially growing cells, are discussed.


Activation and inhibition of the Mg-Ca-ATPase from E. coli by Mg2+ and Ca2+. MgATP of CaATP is the substrate of the Mg-Ca-ATPase. At low Mg2+- or Ca2+-concentrations the ATPase is activated by Mg2+ or Ca2+, the activator being essential for activity. At higher Mg2+- or Ca2+-concentrations the Mg-Ca-ATPase is inhibited competitively. Thus the real Km is smaller than reported in the literature. H+ competes with Mg2+ or Ca2+ for the metal binding sites.


Poncimarin, a new coumarin from Poncirus trifoliata L. A new coumarinic derivative (Poncimarin; C19H22O5; m.p. 140 C degrees) was isolated from unripe fruits of Poncircus trifoliata L. which from spectroscopic evidences was formulated as 7-(2,3-epoxy-3-methyl butyloxy)-8-(2,3-epoxy-3-methyl butyl) coumarin.


Over-production of porphyrins and heme in heterotrophic bacteria. In Escherichia coli, Pseudomonas aeruginosa, and Achromobacter metalcaligenes gamma-aminolevulinic acid synthase and gamma-aminolevulinic acid dehydratase appear to be the rate limiting enzymes of porphyrin and heme biosynthesis. Bypassing of these enzymes by addition of gamma-aminolevulinic acid or porphobilinogen results in overproduction of tetrapyrroles.


Effect of zinc and cadmium on delta-aminolevulinate dehydratase of red blood cells in protecting against enzyme losses during storage. The effect of zinc and cadmium on delta-aminolevulinate dehydratase of bovine erythrocytes stored at -30 degrees C for different times was determined. The results show a. storage of erythrocytes leads to an enhancement of the enzyme activity, which after six weeks is 165% (500 muM ZnCl2) respectively 220% (100 muM CdCl2) for red blood cells of calves, and after four weeks in 420% respectively 450% (same concentrations) for red blood cells of adult cattle, b. the older the samples are, the higher is the metal concentrations, needed for activation.


[Partial synthesis of adenosylcorrinoids and of 1,N6-etheno-adenosylcobalamin with a fluorescent nucleoside (author's transl]. Adenosylcobalamin and several analogs of this corrinoid are prepared by an improved method and separated by an one-step-chromatography on columns of carboxymethylcellulose. Pure water is the eluting agent. The same method is suitable for the preparation of 1,N6-ethenoadenosylcobalamin, a corrinoid with fluorescent nucleoside.


Gamma-radiolysis of aqueous solution of proline. Gamma-radiolysis of 0.05 M neutral proline solution was investigated. As the main reaction of proline radiolysis in oxygenated solutions, the hydroxylation reactions leading to hydroxyproline formation and the reactions leading to the total destruction of the pyrrolidine ring were shown. In oxygen-free solutions the yield of these reactions decreased; in this medium combination reactions giving rise to higher molecular weight combination products (up to approx. 1500) belong to the most important ones. In addition to radiation-decarboxylation reactions the products of radiation-carboxylation reactions were also detected in the irradiated solution of proline free of oxygen.


Interactions between retinal and phospholipids in monomolecular films at acid pH. The surface properties of mixed monomolecular films of retinal and phospholipids (p. lipids) are measured as a function of mole fraction at a nitrogen-water interface. An acid pH of 6.0 is maintained in the aqueous phase. Before irradiation the surface potential deltaV for 9-cis retinal, 11-cis retinal, phosphatidyl serine (PS) and phosphatidyl ethanolamine (PE), at pi=12 dyn/cm, are 490 mV, 645 mV, respectively. Before irradiation, A0 for 9-cis and 11-cis are 58 A2 and 48 A2, respectively. Experimentally measured isotherms are compared with theoretically calculated isotherms. In case of mixed films of retinal and PS the experimental isotherms are greater than theoretical, while mixed films of retinal and PE are smaller than theoretical. A maximum value for the difference between theoretical and experimental areas are obtained at (retinal)/(p. lipid)=0.1. Retinal and p. lipid do not appear to form a Schiff base, charge transfer or any other type of complex at pH 6. A eutectic type mixture between retinal and p. lipid may occur on the surface. A light induced change in deltaV of -130 mV is observed in the case of 11-cis and PE. The significance of these findings with respect to visual excitation is considered.


Binding of antibodies onto the thylakoid membrane. I. Maximal antibody binding and adsorption of antibodies to lipids. The binding of antibodies onto the lamellar system of Antirrhinum majus was determined in dependence on the serum addition. The unspecific adsorption of serum proteins was taken into account or eliminated. The binding of antibodies as a function of the amount of serum added is seem from a saturation curve. From an antiserum obtained by hyperimmunization with stroma-freed chloroplasts, the chloroplasts bind maximally 1 gram antibodies per gram stroma-freed chloroplasts. From an antiserum to the proteins of the thylakoid membrane prepared in the same way an equal amount of antibodies is adsorbed. It is assumed that with this amount the surfaces of the lamellar system accessible to antibodies is completely covered by antibodies. For an antiserum to monogalactosyl diglyceride a maximal antibody binding of 0.16 g, for sulphoquinovosyl diglyceride 0.12 g and for phosphatidyl glycerol 0.13 of antibodies per gram stroma-freed chloroplasts are obtained. The significance of these results with respect to the molecular surface structure of the thylakoid membrane is discussed.


[Mode of action and structure-acitivity-relationships of the aminotriazinone herbicide Metribuzin. Inhibition of photosynthetic electron transport in chloroplasts by Metribuzin (author's transl)]. The influence of the aminotriazinone herbicide Metribuzin on photosynthetic reactions of isolated chloroplasts is investigated. Metribuzin inhibits all Hill-reactions when water is the electron donor, but not photoreductions by photosystem I at the expense of an artifical electron donor. The PI59-value is 6.7. Cyclic photophosphorylation is not affected by Metribuzin. Measurements of the prompt and delayed fluorescence of the photosynthetically active chlorophyll support the notion, that Metribuzin inhibits photosynthetic electron flow between the primary and seondary electron acceptor of photosystem II (Q and plastoquinone). The relationship of inhibitory potency to chemical structure is investigated by comparing a number of related aminotriazinones. The effect of various substituents is discussed.


[Characterization of messenger ribonucleoproteins from yeasts (author's transl)]. A method to produce in 15 min stable sphaeroplasts from yeast of the early log-phase is described. This method was used to isolate polysomes, after specific radioactive labelling of mRNA. The mRNP *-particles, isolated from polysomes, have sedimentation-coefficients from 10 to 80S and a density of 1.4 g/cm3, the polysomes a density of 1.58 g/cm3 and the ribosomes a density of 1.6. g/cm3.


Sensitivity of Escherichia coli to viral nucleic acid, X. Ba2+-induced competence for transfecting DNA. Effect of alkaline earth metal ions on induction of the competence for DNA transfection was investigated. Unlike spheroplasts, the bulk of the bacteria treated with these ions retains colony-forming ability. The order of effectiveness for transfection of phiA replicative-form DNA has been found to be Ba2+ greater than Ca2+ greater than Sr2+ greater than Mg2+. The competence of Ba2+-treated cells is 3 to 5 times higher that that of Ca2+-treated bacteria and about 40 times higher than that of lysozyme-EDTA spheroplasts. The Ba2+-dependent transfection is cryophilic and formation of the infective complex occurs very rapidly at 0 degrees C, But not at 37 degrees C.


[Sequence variation on an antibiotically active tripeptide (author's transl)]. Seven tripeptides with the sequence L-Arg-D,L-X-L-Phe were synthesised. Three of these peptides showed antibiotical activity on fungi. The amino- and the carboxylic terminus of the peptide L-Arg-D,L-Phe-L-Phe, which showed antibiotical activity, were changed to give the sequences L-X-D,L-Phe-L-Phe or L-Arg-D,L-Phe-L-X respectively. The resulting tripeptides showed no antibiotical activity.


[Ultrastructural localization of alkaline phosphatase and ATP-ase in cyst stages of Sarcocystis tenella (Sporozoa, Coccidia) parasitic in the esophagus of sheep (author's transl)]. The activity of adenosine triphosphatase and alkaline phosphatase was investigated at the fine structural level in the cyst stages of Sarcocystis tenella parasitic in the esophagus of sheep. Alkaline phosphatase reaction was observed along the outer membrane of the parasite's pellicle. The enzymatic activity was much higher on the surface of metrocytes than that of zoites, which proved to be infectious. No reaction was noted in the interior of the parasites. However, a significant amount of alkaline phosphatase activity occurred along the inner surface of the 25 nm thick primary layer of the cyst wall. No evidence of the reaction of this enzyme was seen in the secondary cyst wall, which consisted of degenerated host cells. ATP-ase activity was found in a considerable degree along the primary cyst wall (=directly limiting the cyst's interior), whereas the ground-substance of the cyst, surrounding the parasites, is free of deposits. In the parasites ATP-ase was localized in the endoplasmic reticulum, in the perinuclear space, and between the two inner membranes of the three-layered pellicle. Only rarely a slight reaction was seen in the mitochondria of the metrocytes, which are the reproductive cells. The other organelles typical for S. tenella were free of ATP-ase. The results indicate that the enzymes studied participate in the growing process of the cysts, in which finally the infectious zoites remain in a more or less inactive state. The localizations of the enzymes corresponded with the results known from metazoa.


[Resin linked puromycin (author's transl)]. A biospecific resin for the retardation of ribosomal peptodyltransferase by affinity chromatography was prepared by condensing the alpha-amino group of puromycin with the N-hydroxysuccinimide ester of CH-Sepharose 4B to yield the resin-linked puromycin derivative 4.


[Affinity chromatography with 125I-labeled RNA]. Iodination of RNA with carrier-free 125J changes the affinity to cellulose and polyuridylic acid. Because part of the radioactive RNA is tenaciously bound to the respective matrix [125J[RNA cannot be used for affinity chromatography.


Neuraminidase treatment enhances the lysolecithin induced intercellular adhesion of amoeba proteus. Egg lysolecithin induces intercellular adhesion of Amoeba proteus. Pre-treatment of cells with Vibrio cholerae neuraminidase (VCN) increases the adhesive property of cells as was evidenced in their formation of larger cell aggregates than controls. A possible role of VCN exposed receptor sites in cellular adhesion is suggested.


[The pCO2-/pO2-optode: a new probe for measurement of pCO2 or pO in fluids and gases (authors transl)]. To measure pO2 or pCO2 in gases or fluids the fluorescence indicators pyrene butyric acid or beta-methylumbelliferon were used, which were separated from a measuring medium by a gas-permeable membrane. For pCO2 the pH changes in NaHCO3 solution were monitored by methylumbelliferon. Changes in its fluorescence signal, S, were measured at lambda (emmission) = 445 nm. Excitation was brought about by light of wavelength lambda1 = 318 nm (fluorescence signal S1), and lambda2 - 357 nm fluorescence signal S2). Drawing lg(S2/S1) against lg(pCO2-1), one obtains a straight line between a pCO2 of 10 and 70 Torr. The response time is about 3 to 4s (90%). For pO2, pyrene butyric acid is used (lambda(excitation) = 342 nm; lambda(emission) =395 nm). A linear increase in pO2 between 0 and 300 Torr yields a linear increase in the reciprocal fluorescence signal. The response time is about 3 s (90%).


[The effect of dimethyl sulfoxide (DMSO) on the blood circulation in the pulp]. The physiological and pathological regulatory mechanism of the endodontal blood circulation are demonstrated by means of vital microscopy with special respect to the specific vasoactivity of the DMSO. The authors' evaluation of the results includes discussion of the potentially utilizable effects of DMSO, but also the negative sequelae of the carrier effect.


[The enamel-dentin membrane in the scanning electron microscope]. In the scanning electron microscope in detailed structure of the dentino-enamel membrane can be investigated. This membrane is mineralized. After partial demineralization it covers the bottom of prism margins and enables their firm anchoring on the surface of dentin. Here the prism bundles are anchored in dishlike formations, on the bottom of which a network of fibers is found, covered by the dentin-enamel membrane. Through artificial fissures in the membrane the above mentioned network of fibers can be seen.


[The simultaneous surgical correction of lip and nose in unilateral cleft lip and cleft palate]. The advantages of correction of the lip with the method by MILLARD and the own mode of procedure by correction of the nose at patients with unilateral cleft lip is sketched. The possibility of combination of these both operations in one session is demonstrated. It is following the discussion of the superiority of the combined correction of cleft lip and nose.


[Chemical and radioisotope studies of the metabolism in mineralized tissues following use of calcium-, phosphate-, and fluoride-containing toothpastes]. During experimental research work carried out on animals with a view to clearing up the caries-forming processes, metabolic disturbances could be detected in the hard tissues of teeth after administering a cariogenic diet. The aim of the present work was to investigate the effects of tooth-pastes with calcium, phosphate and fluoride content upon changes in metabolic processes, and thus to clarify the mechanism of the anti-cariogenic effect exerted by fluoride tooth-pastes.


[Microbiological characteristics in rats given cariogenic diet following the application of fluoride containing toothpastes]. The paper deals with experimental research work carried out on Wistar rats, regarding the effect of fluoride tooth-pastes in varying compounds on microflora in the oral cavity. The findings show that the most favourable effect on the microbial situation in the Cavum oris is exerted by fluoride tooth-pastes prepared on a calcium-free basis.


[The fluoride content in the crowns of molars following application of fluoride-containing toothpastes]. The paper describes chemical-analytical research into the fluorine content in the coronal parts of molars of rats after fluoride tooth-pastes made up of differing ingredients were used. On the basis of the findings, recommendations are made as to ways of clinically testing the tooth-pastes.


[Sulphated glycosaminoglycans as virus inhibitors. 3rd communication: therapy of viral diseases by means of glycoasaminoglycanpolysulphates. Establishment of fundamentals in experiments with laboratory animals (author's transl)]. Following the in vitro and in vivo demonstration of their inhibitory effect upon 17 D yellow fever virus (Comm. I and II) it has been tried to demonstrate the therapeutic effect of three GAGPS (L1, L5, L8)1 in experimental animals. It had been found that L1 possessed the strongest inhibitory action and L5 the lowest toxicity. L8 served as control substance with different chemical structure. Mice that had been intracerebrally infected with 50 to 100 LD50 yellow fever virus were subsequently treated with L1, L5 and L8 by i.v., i.p., i.m., and oral routes. At first it was found by cytophotometric measurements that the i.c. applicated substances accumulated in the nerve cells of the hippocampus major, the cerebellum (Purkinje cells) and the cortex; the uptake was nearly doubled if a mixture with virus was used (Table 1). Following preliminary experiments to determine the adequate quantity of virus, five experiments were performed in the order mentioned. In the first series were treated groups of 30 animals after intracerebral infection with 100 mug/0.02 ml L1 by the i.m. and i.p. routes respectively, beginning from the first day p.i. for a period of seven days (Table 2). A certain difference of the rate of deaths and surfivals was seen between the treated and untreated groups. Among the treated mice delayed death was a prominent occurrence (Fig. 1). A second experiment involving a double dose of L1a (200 mug/0.02 ml) from another batch of GAGPS showed no better effect (Table 3). An explanation was given by the fact that L1a demonstrated a moderate toxicity with high doses about 5000 mug/ml in the i.c. control (Table 4 and 5). A graphic representation of both experiments can be found in Figs. 2a and 2b. The relative low virus input in the third series as shown in the virus control impedes additionly clearcut results. In the fourth experiment the infected mice were treated with GAGPS doses between 250 and 2500 mug/ml; L1 was administered by the oral, L5 and L8 by the intraveneous route. The death rate of the animals treated with low doses of L1 (250-1000 mug/ml) is diminished clearly and there was a significant difference between treated and untreated mice when L5 and L8 were applied (Table 6). Fig. 3 shows the graphic representation of experiment four. The good results of treatment were confirmed by histopathological findings (Table 7). There was a clear difference in the kind and quantal distribution of cerebral lesions in treated and untreated mice. In the last series L1 was administered by the i.v., L5 and L8 by the oral route (Table 8). Although the virus dose given in this series was rather low a protective effect was seen with low doses of L1 (312 mug/ml) and L5 )500 and 1000 mug/ml). Also these results were confirmed by histopathological examination. In summary, the GAGPS L1, L5 and L8 were found to have a clear therapeutic effect upon the experimental encephalitis of mice caused by infection with 17 D yellow fever virus, in the case of experiment four with statistical significance.


[Use of euphyllin and adrenaline electrophoresis in the sanatorium-health resort treatment of hypothalamic syndromes]. The authors propose a comprehensive treatment of patients with hypothalamic syndromes (vegetative-visceral vascular forms) by a differentiated use of electrophoresis of euphellin. This method was efficacious in expressed headaches with increased blood pressure and cerebral vessels tone, crises of a sypmathico-adrenal character. Electrophoresis of adrenalin was mainly indicated in symptoms of sympatho-adrenal insufficiency and crises of a vago-insular structure.


[Computer analysis of ventricular pressure recordings: evaluation of myocardial contractility on the dog heart in situ]. The contractile indices Vmax (maximum shortening velocity of the contractile element) and ARPD (power averaged rate of power  density generation) which have been shown to be unaffected by alterations in preload and afterload were computed from isovolumic left ventricular pressure data of dogs. The two indices were tested for their ability to detect changes in contractility induced by a positive inotropic drug (Isoprenalin). Whereas a good correlation was found between ARPD and Vmax (coefficient of correlation 0,895) the index ARPD was more sensitive to augmentation of myocardial contractility; also because it is simpler to obtain computationally and more appropriate for the intact heart from a theoretical point of view. ARPD should be useful especially for quantification of acute changes in myocardial contractility.


[Mechanism of synaptosomal degradation of ATP in connection with involvement of adenosine in the transmission process]. In the present paper the mechanism of the adenosine formation by a mixture of nerve ending and transmitter granula fractions was invesitgated. The adenosine formation in vivo is only possible via the whole degradation chain ATP - ADP - AMP - adenosine. The enzymes involved are ATPases, adenylate kinase and 5'-nucleotidase. The ATPase and adenylate kinase effectors Ca++ and Mg++ can be regarded as trigger ions switching on and off the degradation chain. The adenylate kinase represents a key enzyme within the whole chain. In the ion-activated state a non-inhibited adenosine formation was observed, when the initial ATP concentration amounted to less than 0,1 muMol per mg synaptosomal membrane protein. Under these conditions the whole chain velocity is mainly dependent on the 5'-nucleotidase concentration, because ATPases and adenylate kinase remove the nucleotidase inhibitors ATP and ADP spontanously. The conditions for the optimal velocity of the adenosine formation at the synaptic membrane in vivo in all probability are present. A hypothesis for the mechanism of the synaptic adenosine formation in vivo was developed. The importance of this process in respect to the synaptic transmission was discussed.


Demonstration of a Mg-ATPase and a Na,K-ATPase from the arteria carotis communis of the sheep. Besides the Mg-ATPase, a Na,K-ATPase can be demonstrated in different fractions of smooth muscles of the A. carotis communis of the sheep. The highest activity of Mg-ATPase is observed in the heavy microsomal fraction. The Ca-ion may act as a complete substitute for the Mg-ion in the Mg-ATPase. The proportion of Na,K-ATPase is between 10 and 40%, depending on the preparative conditions used in the individual fractions. Fractionated salt treatment (LiBr, KC1, KBr) improved the assay of Na,K-ATPase but increased strength of the Tris-HC1-buffer considerably reduced its activity.


Changes in rat adrenal medulla following delta9-tetrahydrocannabinol treatment. A histochemical study. The effects of acute (10 mg/kg) and chronic 10 mg/kg for 30 days) administration of delta-9-tetrahydrocannabinol (delta9-THC) have been studied histochemically in the rat adrenal medulla, which include total catecholamines, noradrenaline, histometric measurements of adrenal medullary areas, calcium content of the medullary cells along with adenosine triphosphatase (ATPase), acetyl cholinesterase (AChE) and butyryl cholinesterase (BChE) activities. Acute delta9-THC treatment reduced the total catecholamine content (including noradrenaline) of the gland, was accompanied by increased ATP-ase, AChE, BChE activities and increased calcium distribution in the gland. Chronic delta9-THC treatment caused significant hypertrophy of the chromaffin tissue, with decreased total catecholamine content, although noradrenaline containing areas exhibited no notable change. The calcium content and ATPase activity were increased along with a concomitant increase in AChE and BChE activities. Although the changes in adrenal medullary enzyme activities following both acute and chronic delta9-THC treatment are qualitatively similar, marked quantitative increase is noted in the chronically treated groups. The results indicate an increased total catecholamine releasing activity of the adrenal medulla following acute delta9-THC treatment, while chronic delta9-THC administration produces a preferential release of adrenaline.


[Histochemical studies of mucopolysaccharides in the urethral glands of newborns (glands of Littre) (author's transl)]. Histochemical study of urethral glands (Littre's glands) at birth shows, from that time, a notable secretion of mucopolysaccharides. It appears that the properties of the prosecretion are, in part, different from those described in the adult. In fact, in addition to the acid mucins which loose their affinity for Alcian dyes after digestion by neuraminidase and acid hydrolysis, there is an important elaboration of neutral mucosubstances which reacts positively with PAS even if the latter is combined with Alican Blue staining.


Electronmicroscopic study of the effect of 6-an on the sciatic nerve in newborn rats. 12 hrs, 1, 2, and 4 days after i.p. administration of 2, 3 or 5 mg/kg 6-AN (6-Aminonicotinamide) on postnatal day 5, the following findings were made in the sciatic nerve of the newborn rat. In the myelin-forming fibres a swelling between the axolemm and the first inner intraperiod-line of the Schwann cell develops with displacement of the otherwise intact axon. The myelin formation is not impeded. During the 4 days of control after administration of 6-AN there is continual growth of the myelin sheath with an orderly lamellar structure. Using any dose there is no demonstrable alteration in the Schwann cells--with the exception of the inner cytoplasmic tongue--either in the foetal promyelin and mature nonmyelinated fibres. These findings are discussed and compared with the known glia alterations of the spinal ganglia and dorsal radix after the application of 6-AN.


Relationship between heart weight and the cross sectional area of the coronary ostia. Sex variation in the area of the coronary ostia was studied in hearts with a normal weight (heart weight less than 400 g) and hypertrophic hearts (heart weight greater than or equal to 400 g). Also studied was a possible sex difference in the correlation between heart weight and the area of the coronary ostia. The study was performed on 379 post mortem hearts. In the group of hearts with a normal weight the area of the left coronary ostium in both sexes was greater than the right but the difference was statistically insignificant. There was no sex difference in the area of the left coronary ostium, whereas the area of the right coronary ostium was statistically significantly less in women. In the group of hyperthopic hearts the left coronary ostium increased in area with increasing heart weight. The difference in the area of the left coronary ostium between hearts with normal weight and hypertrophic hearts was stastically significant in men, but not in women. The right coronary ostium showed minimal increase in size with increasing heart weight and the difference in both sexes between the two groups was statistically insignificant. In the group of hypertrophic hearts the heart weight was best correlated to the area of the left coronary ostium in both sexes, but the values did not reach statistically significance. On the basis of this study the area of the right coronary ostium appears to be a bottleneck with regard to an adequate blood supply to a hypertrophic myocardium.


Studies elucidating the importance of thymus on the degree of increased blood pressure and vascular disease in renal hypertensive mice. A comparison of the disease in nude and haired littermates. The degree of round cell infiltration around hypertensively damaged heart arteries in one kidney Goldblatt hypertensive mice is more pronounced in haired mice with normal thymus function than in their nude littermates with genetic aplasia of the thymus. The level of hypertension and the prognoses for the hypertensive mice are, however, not influenced by the presence of thymus and thymus derived T cells. The results give evidence that delayed type immune reactions are involved in the hypertensive vascular disease in mice, but fail to support the assumption that they have pathogenic importance for either the level of hypertension or the prognoses of the one kidney Goldblatt hypertensive mice.


Ethanol-induced changes in cation-stimulated adenosine triphosphatase activity and lipid-proteolipid labeling of brain microsomes. The experiments described are based on the hypothesis that prolongation of the depressant action of ethanol leads to compensatory changes in neuronal membrane structures involved in impulse conduction and transmission, and that these become manifest as increased tolerance and withdrawal hyperexcitability. Behavioral tolerance was tested by means of the tilted plane test in rats consuming 9-10 g ethanol/kg/day in a liquid diet fed ad lib., or given 5 g/kg every other day by stomach tube, or doses rising from 6 to 9 g/kg/day maintaining continuous intoxication. All treatments were continued for about three or four weeks before testing. Rats consuming ethanol at a self-regulated rate did not develop tolerance, evidently because sufficient alcohol levels were not built up. Prolonged intoxication induced a high degree of tolerance and withdrawal symptoms, whereas intoxication every other day induced an intermediate degree of tolerance. When no definite abstinence symptoms were associated with the behavioral tolerance, cation stimulated ATPase activity of the brain microsomal fraction was not changed. With increasing withdrawal excitability, there was a relative increase in Na+, K+-stimulated ATPase and an decrease in Mg2+-stimulated ATPase whereas total activity of the enzyme system was not altered. 14C-serine was used as a precursor in order to detect changes in the metabolism of membrane components. So far, only acute experiments have been carried out in vivo. Heavy intoxication (6 g ethanol/kg by stomach tube) inhibited labeling of brain microsomal lipids and proteolipids. In "hangover", proteolipid labeling had returned to the control level whereas lipid labeling was still depressed. Cerebral cortex slices from rats in a withdrawal state after prolonged intoxication, and from control rats, were incubated in vitro with 14C-serine. Unstimulated tissue showed no effect of the prior treatment. When electrical stimulation was applied, much more activity was recovered in microsomal lipids of slices from withdrawal animals than from controls.


Use of the reversed passive Arthus reaction as a test for anti-inflammatory agents. The immune complex-induced reversed passive Arthus (RPA) reaction in the rabbit has been investigated as a screening test for detecting anti-inflammatory agents potentially more effective in the treatment of rheumatoid arthritis than those presently available. RPA lesions, characterized by edema, erythema, and hemorrhage, were elicited by intravenous injections of bovine serum albumin (BSA) followed by intradermal injections of rabbit anti-BSA antiserum. The anti-edema activities of compounds (mg quantities required for testing) were evaluated after their administration by the intradermal route (compounds admixed with antiserum) as well as by the intraperitoneal route. Of 14 reference anti-inflammatory agents tested by the intradermal screening procedure, only aurothioglucose and chloroquine were inactive. Other pharmacologically active compounds (e.g. antihistamines, anti-complement agents, cytotoxic-immunosuppressives) were also evaluated after their intradermal administration. Protoporphyrin, phloretin, and hexadimethrine bromide (Polybrene) were active. When whole antiserum, or the antibody fraction of the serum, was used to eliminate nonspecific edema, intraperitoneally administered reference agents were found to be effective in the RPA test.


Urinary excretion of glycosaminoglycans in malignant diseases of the haemopoietic and lymphatic tissues. A study has been made of the urinary excretion of glycosaminoglycans (GAG) in 50 patients with malignancies, including 6 patients with acute myeloid leukaemia (AML), 11 with chronic myeloid leukaemia (CML), 10 with chronic lymphatic leukaemia (CLL), 10 with multiple myeloma (MM), 7 with Hodgkin's disease and 6 with mycosis fungoides (MF). The total urinary GAG were isolated by precipitation with cetyltrimethyl-ammoniumbromide (CTAB), and assayed in terms of their hexuronic acid content. A statistically highly significant increase in the excretion of total GAG was observed in all the disorders studied, except Hodgkin's disease, the highest value being seen in myeloid leukaemia (ML). Constant amounts of non-dialysable urinary GAG were electrophoresed in 0.5 M lithium acetate on cellulose acetate strips, and stained with alcian blue. The densitometric tracing derived from the electrophoresis strips were analysed with a Du Pont Curve Resolver. The electrophoretic data suggested the existence of a qualitative deviation in GAG excretion in CLL and in MF, in that patients with these diseases excreted on an average larger than normal amounts of slowly migrating GAG fractions. Pooled crude urinary GAG material from patients with CLL, MF, AML and CML and from control subjects was further purified and subjected to analytical studies. These indicated that a similar qualitative urinary GAG distribution exists in ML and in controls, whereas the urinary GAG in CLL and MF patients contained relatively more dermatan sulphate (DS, in terms of iduronate) than those of the controls.


The coronary arteries in Marfan's syndrome. A morphologic study. The coronary arteries of five hearts of patients known to have had Marfan's syndrome were studied microscopically. In four of the five cases the extramural coronary arteries showed structural changes consisting of accumulations of acid mucopolysaccharides and a concomitant disruption of the connective tissue elements. These changes closely resembled the changes that frequently occur in the aorta and that are generally described as "cystic medial necrosis". They are considered to be an important expression of wall weakening. In one case a laceration, probably traumatic, of a coronary artery led to formation of an "incomplete dissecting aneurysm." "Cystic medial necrosis" was found once in the proximal segment of the sinus nodal artery and once in the musculoelastic layer of a coronary artery that reached the atrioventricular node from the ventricular septum by perforating the anulus fibrosus. However, in contrast to findings in a previous report, the intramural coronary arteries were otherwise unaffected. The clinical significance of these data is as yet unknown particularly since these alterations are intramural and initially do not cause narrowing of the lumen. Nevertheless, the possibility of coronary arterial involvement by the metabolic disorder of Marfan's syndrome should always be considered during clinical assessment of this lesion.


Labelling of murine epidermal Langerhans cells with H3-thymidine. Epidermal Langerhans cells may be identified by light microscopy by their strongly positive reaction following incubation for ATPase activity. Intact sheets of epidermis from mice killed at various time intervals following a single pulse label of H3-thymidine were incubated to demonstrate ATPase activity and subsequently processed for autoradiography. In specimens taken one hour after labelling, many basal keratinocytes were labelled but very few ATPase-positive dendritic cells. At subsequent time periods a few pairs of labelled ATPase-positive cells were found but individually labelled cells were not observed. The findings suggest that epidermal Langerhans cells form a very stable (labelling index less than 0.01%) self-replicating population which divides to maintain cell spacing during growth. No evidence was found for migration and interchange of Langerhans cells with the connective tissue, or for an origin of Langerhans cells by transformation of another cell type.


[Contact dermatitis in children].

[Contact dermatitis in children]. After reviewing the frequency differences between the two great eczema varieties in child, in which atopic eczema develops in early childhood and contact dermatitis at age six, this work includes a total of 37 cases of atopic dermatitis observed within a period of three years in a "Pediatric Dermatology Service". Orto-ergic dermatitis (10 cases): Predominance in small children. Provoked by irritating drugs, hygiene products and synthetic clothing. Allergic dermatitis (27 cases): Most frequent at age six. Due to various sensitizing agents, drugs, sulphamids, antibiotics, phenotiazins, clothing, detergent and wild plants. There was a total of 27 cases of atopic dermatitis, divided in two groups in order to facilitate the diagnosis with patch tests. The first group included a total of 12 positives; the most common allergens were plastic materials, drugs, paraphenilendiamine and wild plants. Most of the group corresponded to drug induced dermatitis. A total of 15 cases were considered in the second group yielding doubtful results. In this group and causative agents most commonly involved were drugs and clothing (mainly shoewear).