A fluorescence staining method for the examination of microorganisms on natural substrates. A fluorescence staining technique is described which uses the magnesium salt of 1-anilino-8-naphthalene sulfonic acid as a protein stain. At a concentration of 2.5 or 3.0 mg ml-1 in distilled water or phosphate buffer the compound does not fluoresce under ultra-violet or far blue illumination until it is bound to a protein or similar compound. It can be applied to natural substrates such as soil, food materials, organic material in water, etc., and can be examined immediately without any washing procedures. The application of this staining method to detect microorganisms on dead green algae (detritus) is described.


Techniques and applications of autoradiography in the light and electron microscope. Principle, examples of application and technical details of autoradiographic procedures are described. Tested recipes for light microscopic autoradiography of biological material and for qualitative electron microscopic autoradiography are given. Problems of quantitative autoradiography are discussed.


Orseillin BB and crystal violet, a differential staining combination for paraffin sections and water mounts of fungi. A saturated solution of orseillin BB in 3% acetic acid followed by a 1% aqueous solution of crystal violet provides an excellent differential staining for sections of ascomycetous fructifications. The technique stains a wide variety of fungus and host cells, revealing considerable morphological and cytological detail. It is appropriate for microscope slides both of unfixed material mounted in water and of picric acid-fixed paraffin sections.


Combination chemotherapy of advanced previously treated Hodgkin's disease with streptozotocin, CCNU, adriamycin and bleomycin. Seventeen patients with advanced, previously treated Hodgkin's disease received therapy with a combination of streptozotocin 500 mg/m2/day i.v. days 1--5, CCNU 100 mg/m2 orally day 1, adriamycin 45 mg/m2 i.v. day 1, and bleomycin 15 mg/m2 i.m. days 1 and 8 at 28-day intervals (SCAB). The overall response rate was 59% with six patients (35%) achieving complete remission and four patients (24%) entering partial remissions. No maintenance therapy was given and the median duration of complete remission was 8+ months (range 2+-18+ months), while the median duration of partial remission was only 2 months (range 2-3 months). The median duration of survival from the start of therapy for the complete responders was 16+ months (range 5+-25+ months) while the median survival for the partial and nonresponders was only 5 months (range 2-13 and 3-11+ months, respectively). Toxicity was a major problem with this drug combination. Myelosuppression occurred regularly and was severe after 25% of courses. There were two deaths directly related to drug-induced myelosuppression. Other serious toxicities included bleomycin-induced pulmonary toxicity in three patients, with one death; renal tubular dysfunction secondary to streptozotocin in three patients; hepatic dysfunction in three patients and severe weight loss in three. SCAB has proven to be an active although toxic combination which is not cross-resistant to MOPP-type regimens. Alterations in drug dosages and scheduling are being evaluated in an effect to ameliorate toxicity and preserve efficacy.


Cyclophosphamide, vincristine, and prednisone (CVP) versus adriamycin, bleomycin, and prednisone (ABP) in stage IV non-Hodgkin's lymphomas. In stage IV non-Hodgkin's lymphomas, CVP (cyclophosphamide, vincristine, and prednisone) was randomly compared to ABP (adriamycin, bleomycin, and prednisone). Of 62 patients entered into the study, 57 (CVP 27, ABP 30) were considered evaluable for comparison. In patients with liver and/or marrow involvement a second biopsy was performed to define complete remission (CR). CR occurred in 48% of patients treated with CVP and in 50% of those given ABP. The median duration of CR was 10.5 and 20.5 months, respectively. The difference is not statistically significant. Also, the survival of complete responders was not significantly different between the two treatment groups. After cross-over, secondary treatment with CVP produced an overall response rate of 40% (six of 15), compared to 50% (six of 12) obtained with ABP. In the ABP group, four patients developed a reversible interstitial penumonia. In two other patients, cardiomyopathy (fatal in one) was observed. In conclusion, although complete remission was similar in both groups, cumulative toxicity occurred in few patients given ABP. However, this combination could represent an effective alternative treatment to be used either in CVP failures or sequence with CVP.


Epidemic measles in a highly vaccinated population. During November, 1975, to May, 1976, measles occurred at a rate of 20.3 cases per 1000 in a purported immunized population, of whom historical and serologic survey revealed that 9 per cent had no history of either measles illness or vaccination and 18 per cent did not have detectable measles antibody. Antibody was detectable in 92 per cent of those vaccinated at greater than or equal to 13 months, 80 per cent at 12 months and 67 per cent of those vaccinated when less than one year old (P less than 0.001), but no significant differences existed with increasing years since vaccination (P greater than 0.1). A second vaccination increased detectable antibody prevalence only in those originally vaccinated when less than nine months old (42 to 80 per cent, P less than 0.02). During a measles outbreak, more cases occurred in those receiving vaccine when less than 12 months old than in those vaccinated at greater than or equal to 12 months (37 per cent vs. 9 per cent, P less than 0.001). A second vaccination protected those originally vaccinated at less than 12 months (35 per cent ill without a second vaccination vs. 2 per cent with, P less than 0.001). Thus, a single measles vaccination of children less than 12 months old does not protect; a second vaccination will protect this group.


Chemical modification of DNA polymerase phosphoprotein from avian myeloblastosis virus. Fractionation of purified avian myeloblastosis virus DNA polymerase, after phosphorylation in vitro, revealed the presence of a small acidic proten, a phosphate acceptor polypeptide with high specific activity. Its presence in the phosphorylated form with the polymerase resulted in as much as a 10-fold increase in the rate of DNA synthesis. Its presence in the dephosphorylated form with the polymerase had no effect in the rate of DNA synthesis.


[Diagnosis of primary and secondary tumors of the liver]. A brief epidemiological and clinical introduction is followed by an examination of the most useful methods for the diagnosis of primary and secondary tumours of the liver: alpha protein values, scintiscanning, arteriography of the coeliac artery, splenoportography, and laparotomy with directed biopsy. The diagnostic specificity and percent positivity of each method are illustrated.


Chemotherapeutic retroconversion of immature teratoma of the ovary. There has been a growing interest in treating malignant germ cell tumors of the ovary with adjuvant chemotherapy following appropriate surgery. This is a report of 3 patients treated in this manner. Of interest is the finding in all 3 patients of residual mature metastatic teratomatous tissue representing many tissue types. The concept of possible chemotherapeutic "retroconversion" or "in situ destruction" of immature tissue may be the mechanism for this result. All patients are alive and presumably well at the time of this report.


Cell-mediated immunity to herpes simplex virus types 1 and 2 antigens in leukoplakia and carcinoma in man. Cell-mediated immune responses to herpes simplex virus 1 and type 2 virion and non-virion antigens were assessed in patients and in controls with oral leukoplakia, carcinoma and recurrent herpes labialis. Enhanced proliferation was found in lymphocytes from patients with recurrent herpes labialis or with leukoplakia showing epithelial atypia, and depressed responses were found in carcinoma. Very significantly positive correlations were shown between the responses to each of the herpes virus antigens. A specific increase in cell-mediated immunity to herpes virus in epithelial atypia was confirmed, but separation of the nonvirion from virion antigen is necessary before specific cell-mediated immune responses to the non-virion antigen can be assessed.


The synthesis of a variant-specific antigen by Trypanosoma brucei in vitro. A variant-specific surface antigen from a cloned population of Trypanosoma brucei S42 has been isolated and partially characterized. [35S]L-methionine was found to be incorporated into this material by cells incubated in vitro in a chemically defined medium. Incorporation of [35S]L-methionine was inhibited by cycloheximide and puromycin at concentrations which are known to specifically inhibit protein synthesis in other systems. The rate of synthesis of the variant-specific antigen in vitro has been estimated to be about 8% of the rate in vivo. Newly synthesized [35S]L-methionine-labelled variant-specific antigen was incorporated into the surface coat.


Identification of squamous cell carcinoma of the head and neck by tissue culture and immunological testing. We outline the techniques used to successfully grow squamous cell carcinoma in tissue culture, and to test the cellular immunity of the patient by lymphocyte cytotoxicity studies. Lymphocytes cultured with malignant squamous cells killed from 40 to 60 percent of the tumor cells during 48 hours of incubation. These same lymphocytes did not show any killing potential against cultured melanoma cells or against cultured fibroblasts. This demonstrates an immune response that is tumor-antigen specific. There was no evidence of any serum-blocking factor, because the killing potential of these lymphocytes was not significantly altered by the addition of the patients' sera. The implications and potential for early diagnosis made possible by these techniques are discussed.


Tissue alpha-globulins in keloid formation. The deposition of alpha-1-antitrypsin and alpha-2-macroglobulin, both known to be inhibitors of human skin collagenase, is significantly increased in keloids and in hypertrophic scars (as compared to normal skin). However, following intralesional triamcinolone treatment, a marked resorption of these abnormal scars occurs along with a significant reduction of the alpha-1-antitrypsin deposits. These findings suggest that alpha-globulins are involved in abnormal scar formation, and that triamcinolone may remove collagenase and/or protease inhibitors--thereby allowing activation of the collagenase with subsequent breakdown and resorption of the excessive collagen.


[Cerebrospinal fluid diagnosis of multiple sclerosis]. Starting from the knowledge accumulated with respect to the etiopathogenesis and the components of immunoreaction a minimal to maximal program by steps has been developed for the cerebrospinal fluid (CSF) in multiple sclerosis (MS). It is based on fundamental methods (I), special supplementary methods (II), and relatively specific immunological methods (III). For MS five possible conditions of the CSF could be determined from alterations of cells and variations in protein: a (1) typically complete and (2.) typically incomplete immunoreactive encephalomyelitic (encephalitic) syndrome, a (3.) nonspecific CSF-syndrome of low degree and less typical character (in the sense of an acute or subacute irritation syndrome) an (4.) atypical syndrome of a considerable degree, and a (5.) normal condition of the cerebrospinal fluid. The significance of immunoreactive cerebrospinal fluid syndromes to the diagnostic criteria of multiple sclerosis as well as further relatively disease-specific methods (such as the MEM test and MSF assay) of determining cellular immunity, are discussed.


[Evaluation of the relations between serum proteins electrophoresis and other laboratory tests in monoclonal gammopathies (author's transl)]. We have considered interesting to determine monoclonal gammopathies incidence, in 2191 serum proteins electrophoresis performed in our laboratory from January to December 1974. We have found 15 cases of monoclonal gammopathies, some cases combined with Mieloma (3 cases), some other with other with non specific diseases. We have considered the relations between type of gammopathy and other laboratory tests useful for any other diagnose: they are: immunochemical analysis, E.S.R., red and white count, total proteins, Bence Jones protein.


Increased alpha-fetoprotein biosynthesis in rats following DL-ethionine and carbon tetrachloride injuries. The incorporation of 14C-leucine into alpha-fetoprotein (AFP) in serum and liver extracts from DL-ethionine and carbon tetrachloride (CCl4)-injured rats was measured directly by using an immunochemical technique. The maximum value with nearly 10-fold increases of serum AFP was obtained in 3 days of both treatments. However, no significant increase in the immunoprecipitable AFP in liver extracts was detected under these conditions.


[Transvesical prostatic adenomectomy without urethral catheter]. The results are presented, of trans-vesical prosthatic adenomectomy without urethral probe in 40 patients. The drainage of the bladder was achieved by vesical counter-incision, according to the technique suggested by Rose and Novak. A decrease by approximately 70% of the post-operative urinary infections, was obtained.


Hematoporphyrin-sensitized photodynamic inactivation of viral RNA-dependent DNA polymerase. Visible light (6328A) was shown to inactivate RNA-dependent DNA polymerase activity of Moloney mouse leukemia virus, in the presence of hematoporphyrin. The same treatment on exogenous, template-initiation complex had no effect on enzyme activity. It was suggested that this enzyme may be the target of photodynamic inactivation of this and related viruses.


A modified technique for combination of cytochemistry and radioautography of haemic cells. Cultured mouse bone marrow and human blood cells were labelled with 3H-thymidine, smeared, and stained, using peroxydase reagents or the May-Grünwald Giemsa procedure. The slides were dipped in a approximately 12% solution of polyvinylchloride, bleached by a photographic fixer and processed for liquid emulsion radioautography. This method yielded technically very satisfactory radioautographs of pre-stained smears.


[Is there any bimodality of the age distribution of male mammary cancer (author's transl)?]. The authors study the pattern of the curve concerning breast cancer incidence in males as a function of age. The demonstration is made that such a curve is bimodal and, more precisely, could be satisfactorily considered as being made up of the juxtaposition of two normal distribution curves. The interval thus individualized between the mean age of both groups is remarkably steady between 17 and 20 yrs. The phenomenon is not blurred by regrouping the data and the patients population could be divided into two subgroups: one with a mean age of 147 yrs, with a standard deviation of 9 yrs, and bearing on 246 patients, the other one including 1.100 patients averaging 66.5 yrs old with a standard deviation of 9.9 yrs.


Light green crystals in May-Grünwald and Giemsa-stained bone marrow macrophages in patients with myeloid leukaemia. A patient in a terminal stage of myelomocytic leukaemia had a number of elongated crystals in many bone marrow macrophages. The crystals were up to 20 micron long, showed a characteristic light green colour with May-Grünwald & Giemsa, but did not stain with Sudan Black B or PAS. Electron microscopy showed that many of the crystals had a rhomboid form, but failed to show periodicity with the magnification used. A search was made for similar light green crystalline inclusions in patients with different types of leukaemia. Identical inclusions were found in 2 out of 5 other cases of myelomonocytic leukaemia, and in 3 out of 5 cases of chronic myeloid leukaemia. In contrast, such inclusions were not found in any of 6 patients with acute lymphoblastic leukaemia, nor in 4 patients with chronic lymphocytic leukaemia.


In vitro platelet stain preventing (degranulating) effect of various substances, Study of mechanism of action. A few years ago a patients, T.L., with metastatic leiomyosarcoma found to have a serum and tumor factor which caused the granulomere of normal platelets to disappear as seen using light microscopy. The present report describes the effect on platelet granules of several different substances using various anticoagulants. Ultramicroscopy showed in all instances that the platelet stain preventing effect as seen in May-Grünwald & Giemsa stained films reflected a degranulation. Ultramicroscopy was little better than light microscopy in the differentiation between the mechanisms of the various degranulating substances. Different conditions such as variation in anticoagulant or combination of anticoagulants, storage at room temperature for 24h, heating to 56 degrees C for 1 h on the other hand were very useful methods for differentiating between the various degranulating substances. These methods failed, however, to differentiate between patient T.L.-serum and patient T.L.-tumour fraction SSS, probably because the active factor was the same in both.


Neurophysin biosynthesis: conversion of a putative precursor during axonal transport. [35S]Cysteine injected adjacent to the supraoptic nucleus of the rat is rapidly incorporated into a 20,000-dalton protein that, in time, is converted to a 12,000-dalton labeled protein, neurophysin. This putative precursor of neurophysin appears to be synthesized in the supraoptic nucleus and transformed to neurophysin and related peptides during axonal transport to the neurohypophysis.


Severe antithrombin III deficiency in an infant associated with multiple arterial and venous thromboses. Inherited antithrombin III (AT-III, heparin cofactor) deficiency is a rare condition, presenting with thrombotic disease in adult life. This paper reports an 8 months old South African Black male infant with multiple large vessel venous and arterial thromboses, and E. coli septicaemia. This was associated with an extremely low plasma AT-III level. Micronodular cirrhosis and intracytoplasmic hyaline globules in the liver cells were present. These globules were eosinophilic, and PAS-positive after diastase. They measured approximately 5 mu to 30 muin diameter, occurred singly in the liver cells and were located mainly in the periportal areas. The histological findings in the liver are similar to those observed in alpha 1-antitrypsin (AAT) deficiency in which the intracytoplasmic globules represent accumulation of altered AAT. Immunochemical studies carried out on formalin fixed tissue failed to detect cross reaction material with anti-alpha 1 antitrypsin or anti-AT III antiserum. This is the first case report of AT-III deficiency presenting in infancy. It is also the first case associated with distinctive liver pathology. The available data presented are insufficient to distinguish between an inborn defect and acquired caused of the severely depressed AT-III plasma level and the distinctive liver pathology.


Inactivation of alpha- and beta- thrombin by antithrombin-III and heparin. Inactivation of alpha- and beta-thrombin by antithrombin-III and heparin was studied, since it had been suggested that two forms of thrombin exist with respect to heparin sensitivity (Machovich 1975b). It was found that the inactivation rates of alpha- and beta-thrombin by antithrombin were different, namely alpha-thrombin was more sensitive to antithrombin than beta-thrombin. Heparin facilitated the complex formation between alpha-thrombin and antithrombin-III, whereas beta-thrombin inactivation was only slightly affected. Furthermore, heparin protected alpha-thrombin against the inactivating effect of heat, while beta-thrombin lost its activity during the heat treatment. These findings suggest that the formation of beta-thrombin in blood circulation may have an important role in thrombosis predisposition.


Prothrombin determination by means of a chromogenic peptide substrate. A two stage method to determine prothrombin with the chromogenic peptide substrate benzoyl-phe-val-arg-p-nitroanilide has been worked out. Citrated plasma (10 mul) was diluted in 600 mul tris buffer, pH 8.2, ionic strength 0.18 and activated with 250 mul of a commercial rabbit brain-lung thromboplastin. After 325 s incubation at 37 degrees C 200 mul of a 1 mM solution of the chromogenic substrate was added and the increase in absorbance was recorded in a LKB-Beckman 8600 enzyme analyzer. A reading time of 1 minute (including a delay of 20 s) was used which permitted 55 analyses per hour to be carried out. An approximate linear relationship was found between delta A/min and dilutions of normal plasma in prothrombin deficient plasma. The method is insensitive to variations of factors V, VII and X. Less than 10% or normal plasma was needed to "normalize" plasmas deficient in factor V or VII or X. A group of 99 dicoumarol treated patients and 23 normal subjects has been investigated using the present method and compared with a factor II-VII-X determination method. A correlation coefficient of 0.95 was found.


The reversible antithrombin activity of incubated plasma. The thrombin time of normal citrated plasma is progressively prolonged on incubation in open glass tubes at 37 degrees C. The phenomenon is dependent on the temperature and duration of incubation, on the pH, and on the concentration of calcium ions present. Platelet-rich citrated plasma fails to exhibit augmented antithrombin activity when similarly incubated, and the addition of washed platelets to platelet-poor plasma inhibits this activity. The clot retarding action of incubated plasma against thrombin is also manifested against Arvin (Ancrod), but not against Reptilase-R. This thrombin time lengthening may be inhibited by incubation with anti-antithrombin III antiserum thus indicating that the phenomenon of thrombin time lengthening is consistent with enhanced activity of antithrombin III. It is unlikely that alterations in the activity of alpha2-macroglobulin, is important in the reduced thrombin-coagulability of incubated plasma. Interference with the polymerisation of fibrin monomers by the physico-chemical changes may contribute to the observed phenomenon.


Comparison between British Comparative Thromboplastin (BCT) and a factor II-VII-X determination method (simplastin A) based on fresh plasma samples from dicoumarol-treated patients. Comparisons between British Comparative Thromboplastic (BCT) and a factor II-VII-X determination method--Simplastic A--have been carried out monthly during the time from December 1973 to March 1976. On each occasion fresh plasma from 20 dicoumarol-treated patients was analyzed with the two methods. It was found that the linear regression model is well suited for the comparison, that the BCT thromboplastin did not change significantly during the study, and that the method of comparing has a coefficient of variation of approximately 6-8%. This means, for instance, that a ratio of 3.8 with BCT corresponds to a ratio of 3.5 +/- 0.26 with Simplastin A. Thus in one period you may have ratio 3 and in another ratio 4 as the lower limit of the therapeutic range although you use the same thromboplastin and the same method. The importance of having a stable reference thromboplastin and an established method of comparing thromboplastins is stressed.


HLA phenotype and haplotype frequencies in a sample of the Spanish population. The frequencies of HLA antigens and genes were determined in 491 unrelated individuals from the area of Madrid. In order to obtain a fairly good idea of the HLA antigen frequencies in the Spanish population, our results have been compared with those reported from Barcelona. Haplotype frequencies were observed in 212 parents and compared with the frequencies estimated from the 491 individuals. The present data show certain characteristics of the Spanish population and are in good agreement with other studies in the demonstration of linkage disequilibrium between genes of the two series.


Reactivity of anti-HLA sera against mouse lymphocytes. The reactivity of anti-HLA hyper-immune sera was tested by the microlymphocytotoxicity technique (LCT) against a panel of mouse lymphocytes. Three levels of reactivity were observed: negative (immune sera titre 1:20 or less), weak (titre 1:40 to 1:80 with a percentage of dead cells less than 50%) or strong (titre 1:60 or more with 100% killing). Thirteen normal human sera were non-reactive. Eleven out of 60 hyper-immune sera were strongly reactive. Tests using congenic lines showed that the reactivity was controlled at the H-2 complex. One serum (CODRON) was studied in detail. When tested against a panel of strains carrying 10 different H-2 haplotypes, it reacted strongly against lymphocytes H-2d, ja, k, p and r; and did not react, or at least only weakly, against lymphocytes H-2b, f, q, s and v. Tests using mouse strains carrying the recombinant H-2 haplotypes h4, i5, i, y2, g, g2 and t1, suggested that the observed reactivity was directed against structures controlled at the K end of the H-2 complex (H-2.47?). Absorption-elution experiments with human and murine lymphocytes and platelets confirmed that the structures recognised by serum CODRON were determined at the major histocompatibility complex.


Distribution of HLA antigens in a Cuban population. The frequencies of the HLA antigens were determined in a population of 160 unrelated Cubans from three different ethnic groups (white, mixed-blood and negro). We typed for antigenic specificities of the A and B loci, and also for the Cw1 specificity of the C locus. Phenotype and gene frequencies for the different ethnic groups are presented for comparison. The phenotype distribution, the haplotype frequencies for the white group and the genetic distance between this group and a Spanish population were calculated.


HLA-B27 in rheumatoid arthritis and amyloidosis. To study the role of genetically determined immune responsiveness in the pathogenesis of systemic amyloidosis complicating rheumatoid arthritis the HLA antigens were identified in 26 patients with rheumatoid arthritis complicated by secondary amyloidosis, in 44 patients with rheumatoid arthritis, and in 11 patients with secondary amyloidosis of non-rheumatoid origin. Subjects with ankylosing spondylitis, sacroiliitis without peripheral polyarthritis, Reiter's disease, reactive arthritis, erosive osteoarthritis, psoriatic arthropathy, systemic lupus erythematosus or arthritis associated with a gastrointestinal involvement were excluded from the study. Patients with amyloidosis secondary to rheumatoid arthritis had a high frequency of the HLA specificity B27 and of the haplotype likely to bear A2, B27. The association with B27 was closest in the group of male patients with amyloidosis whose rheumatoid arthritis had begun at an early age and who lacked demonstrable rheumatoid factor in serum. These patients may represent a genetically determined subentity of rheumatoid arthritis.


Vertebrate Golgi complexes have beads in a similar position to those found in arthropods. Insects and other arthropods have bead-like structures in Golgi complexes from all cell types. They are arranged in rings at the base of transition vesicles located near the smooth surface of the rough endoplasmic reticulum making the forming face of the Golgi complex and are only seen easily after staining in bismuth salts. Procedures used to demonstrate the beads in arthropod Golgi complexes do not selectively stain any structures where they would be expected to occur in several mouse and tadpole tissues. However, a faint pattern similar to the arthropod GC beads can be made out in the large GCs concerned in the formation of acrosomes during mouse spermatogenesis. Uranyl staining shows particles of about the same size and spacing as the beads of arthropod GCs. We conclude that vertebrate GCs may have beads that differ from arthropods in their staining properties.


Rat iso-alpha1-fetoproteins. Purification and interaction with estradiol-17beta. The combination of polyacrylamide gel electrophoresis and Concanavalin-A-Sepharose affinity chromatography has permitted the isolation on a preparative scale, of four molecular forms of rat alpha 1-fetoprotein: a "slow" and a "fast" fraction, each separable into Concanavalin-A-adsorbed ("high carbohydrate", i.e. rich in accessible alphaD-Mannosyl and alphaD-Glu-cosyl residues) and a Concanavalin-A-non adsorbed ("low carbohydrate") fractions. These four iso-alpha 1-fetoproteins (iso-AFP) bind estradiol-17beta. However, they disclose differences in both their association constants and number of binding sites for this hormone. Very high affinity sites (10(9) are mainly located on the "slow-low carbohydrate" form. Low affinity, high capacity sites are preferentially located on the "high carbohydrate" form. These results confirm the molecular and functional heterogeneity of rat AFT and suggest that the carbohydrate moiety of the protein may have a role in estrogen-AFP interactions.


Human schistosomiasis: Schistosoma mansoni antigen detection in renal glomeruli. Twelve kidney, five biopsy and seven necropsy specimens, all from schistosomiasis mansoni patients were studied by light and immunoflurescent microscopy in an attempt to detect antigen in the glomerular walls. Deposits of IgM, IgG,I gA, IgE, complement C3 and fibrinogen were observered in most cases. Antigen was successfully detected in two cases(one biopsy and one necropsy specimen), both exhibiting proliferative glomerulonephritis. The only clinical manifestation was a slight proteinuria. IgG antibodies eluted from the sutopsy kidney homogenates showed specific binding mostly to Schistosoma mansoni gut, thus spggesting that the fixed antibodies (eluates) are, at least partially, consituted by antibodies similar to the anti-circulating antigen. These data reinfroce the hypothesis that renal injury in schistosomiasis is mediated through an immune complex disease.


Phosphorus-32 for intractable pain in carcinoma of prostate. Analysis of androgen priming, parathormone rebound, and combination therapy. Thirty-three patients with intractable pain caused by diffuse osteoblastic metastases from carcinoma of the prostate were treated with phosphorus-32 (32P) therapy either androgen priming, parathormone rebound, or a combination of both priming methods. Significant response to pain was achieved in 12 of 19 patients receiving testosterone-potentiated therapy, 0 of 5 patients treated with parathormone alone, and 6 of 9 patients receiving a combination of both priming modalities. It is concluded that androgen priming alone is the simplest and most effective method to be used when 32P therapy is being considered for palliative control of pain in patients with carcinoma of prostate.


Microscopic foci of cancer in prostatectomy for benign disease: diagnostic and surgical considerations. A study of 66 patients in whom microscopic foci of prostatic cancer was found in prostatectomies done for benign prostatic hyperplasia revealed in 32 of the 66 cases (48%) that there were in retrospect abnormal rectal findings that should have suggested the possibility of neoplasia. Other abnormalities were elevated serum acid phosphatase (5 patients), asymmetrical enlargement of one lateral prostatic lobe at cystoscopy (7 patients), and unilateral hydroureteronephrosis (7 patients). Urinary retention was the presenting symptoms in 56% of the patients. Difficult enucleation was noted in 41% of the open prostatectomies. The importance of performing careful prostatic biopsy in any suspicious prostatic enlargement is again stressed.


Treatment of benign prostatic hyperplasia with hydroxyprogesterone-caproate: placebo-controlled study. A placebo-controlled study with progesterone compound, 17-alpha-hydroxyprogesterone 17-n-caproate (Primostat), in 39 patients with benign enlargement of the prostate is reported. Statistical analysis of the results showed no evidence of significant improvement in patients receiving hydroxyprogesterone-caproate. No evidence of an effect as compared with the placebo was found when the residual urine, prostatic size, and histologic and ultrastructural changes of the removed prostatic gland in 6 of the patients, and in the luteinizing hormone, follicle-stimulating hormone, and estrogen urine levels in 21 patients were examined. Subjective effects, when carefully analyzed, provided some beneficial evidence, however not substantiated, when the patients' mode of voiding was carefully watched. The reported beneficial subjective improvement might be attributed to the enhancement of the beta-adrenergic response by the progesterone compound of the adrenergic receptors in the posterior urethra and bladder, presumably causing relaxation of its smooth muscle. The problems associated with the choice and measurement of parameters to be used in this type of investigation are discussed, and the absolute necessity of proper controls, statistical analysis, and close follow-up of the patients is pointed out.


Serum alpha fetal protein in a three year old child with hepatoma. A 3 year old child with primary hepatocellular carcinoma and high AFP concentrations is described. Following hemihepatectomy, a sharp decrease and return to normal of serum AFP concentrations indicated the completeness of the surgical procedure. Repeat-normal serum AFP concentrations (less than 19 ng/ml), found during a three year follow-up, correlated well with the absence of clinical, laboratory and x-ray evidence of tumor recurrence. The differential diagnosis of abnormal AFP concentrations in childhood is discussed, and the importance of the AFP assay in the follow-up of post-hemihepatectomy patients for the assessment of the completeness of the surgical procedure, the prognosis, and the early detection of tumor recurrence is stressed.


Treatment of multiple superficial tumors of bladder with intravesical bleomycin. Twenty-six patients with superficial Stage O or A transitional cell carcinoma of the urinary bladder, whose lesions were not amenable to transurethral resection, received bleomycin sulfate intravesically at weekly intervals for eight treatments. Five different drug regimens were tried, and the optimal concentration appeared to be 60 units dissolved in 30 cc. of sterile water. Serum determinations failed to reveal any significant absorption. There was a 27% complete response rate in patients with small tumor burdens. An additional 9% had partial responses which allowed the tumors to be readily managed transurethrally. However, no patients with extensive superficial tumor showed complete response to therapy. Although belomycin used intravesically is active against transitional cell carcinoma, the current cost of the drug precludes its routine use and restricts it to situations in which other agents are contraindicated.


Light and electron-microscopic studies on guinea pig hearts after perfusion with nitro-blue tetrazolium (NBT). Guinea pig hearts isolated by the Langendorff technique were perfused with a nitro-blue tetrazolium salt solution. In spite of a homogeneous dark-blue staining at the macroscopic examination, light-microscopic studies showed a spotty staining of the myocardial fibers. Since no microscopic alterations were found which could explain the unstained areas, ultramicroscopic studies were undertaken. At ultrastructural level, striking alterations of myocardial cells were observed. They consisted of mitochondrial lesions, alterations of the transversal tubuli, and intercalated discs. Arteriolar narrowing and capillary edema were found, suggesting that pathological changes in the microcirculation could be a possible reason for the spotty staining of the myocardial fibers. Some speculations on the nature and localization of the formazan granules are also reported.


[Alpha-2-macroglobulin in children with glomerular diseases (author's transl)]. The serum and urine levels of alpha-2-macroglobulin (alpha2-MG) was determined in 33 children with glomerular diseases and in 26 healthy control children. Healthy children showed a minimum level of 275 mg% and maximum level of 337 mg%, with a mean concentration of 301 mg% and a standard deviation of 13 mg%. No alpha2-MG was detected in the urine. Steroid-treated patients with idiopathic nephrotic syndrome displayed elevated inhibitor levels of up to 490 mg%. This might be a direct result of steroid therapy or a consequence of reactively-increased protein synthesis in response to the renal protein loss. In all these patients the urine was found to be alpha2-MG-negative, irrespective of the presence or absence of proteinuria. In the miscellaneous group of glomerulopathies without the nephrotic syndrome, serum levels of alpha2-MG were shown to be normal. The urinary concentrations of alpha2-MG were related to the activity of the disease. alpha2-MG determination in serum and urine seems to be a tool for differential diagnosis and prognosis in some cases of glomerular disease.


[Dissociation of maturity: a typical variant of motor development (author's transl)]. 71 cases are described of a typical variant of motor development, characterized by dissociation of maturity between the upper and the lower extremities. The affected children develop head and trunk control at the normal time, as well as skill in the use of their arms and hands, but they refuse to stand and walk well into their second year of life. Further characteristic features of this developmental type are: slender limbs generalized reduction of muscular tonus with well-preserved strength and lively spontaneous movements, mental irritation with temper tantrums and stereotype movements. Intellectual development is normal or moderately retarded. The following causes are discussed: constitutional and genetic factors (girls predominate), cerebral damage and treatment with hip abduction splints. The developmental type simulates cerebral palsy, chiefly spastic diplegia. There are only rare descriptions of this condition in the literature. Precise diagnosis is of practical importance, since despite similarities to spastic diplegia, the theurapeutic consequences are quite different. The prognosis of "dissociation of motor development" is good, at least with regard to the neurological symptoms. Normal standing and walking are achieved at the latest by the end of the second year of life. the prognosis of intellectual development depends on the cause and should be individually assessed.


[Investigations of the effect of midodrine on carbohydrate and fat metabolism with particular reference to the diabetic metabolic state (author's transl)]. Midodrine, which is used in the treatment of hypotensive circulatory distrubances was investigated with respect to possible effects on carbohydrate and fat metabolism in 5 healthy subjects and 7 patients with disturbed glocuse tolerance. An i.v. glucose tolerance test was carried out on both groups and was repeated a few days subsequently with simultaneous administration of midodrine (5mg i.v.). Midodrine had no significant effect on glucose tolerance in either group, nor was there any significant effect of midodrine on FFA, serum insulin, triglyceride or cholesterol levels. 15 diabetic patients controlled by different therapeutic regimens (5 by diet only, 5 by oral preparations and 5 by insulin treatment) were given 3x5mg midodrine  orally over a 5-day period and the effects on diabetic control and metabolic parameters compared with a 5-day pretreatment period without midodrine. Midodrine did not cause any change in the quality of diabetic control nor any significant alteration in serum lipid or uric acid levels.


[Trypsin-inhibitory - capacity in humans to respirable allergens or irritants (author's transl)]. The trypsin-inhibitory-capacity was measured with a gelatine-film-technique after JAMES et al, in 520 persons suffering from complaints of upper and/or lower airways in connection with exposure to respirable allergens and/or irritants. As a control group 214 blood donors were examined. After determination of normal range, relations to age, sex, smoking habits, ventilatory parameters, bronchial reactivity, sedimentation rate of blood cells, alpha1-globuline and IgE were examined. In workers exposed to enzymes mean values to trypsin-inhibitory-capacity were found to be in normal range. Determination of trypsin-inhibitory-capacity is proposed to be used in examinations of workers who are to be exposed to enzymes.


[The immunologic tumor diagnosis].

[The immunologic tumor diagnosis]. Issuing from a possibility of classification of tumour-associated antigens (TAA), corresponding to themodern state of recognition, the present situation of the immunological tumour diagnostics is discussed. The direct proof of tumour-associated antigens in the blood or in other body fluids, as one of the fundamental possibilities, is hitherto practically matured only for the alpha-fetoprotein and in this case only contribute to the diagnostics of liver carcinomas and malignant teratoblastomas. The determination of the carcinoembryonal antigen (CEA) in the serum or plasm has at present no essential diagnostic significance, may however, be used for the control of the therapeutic effect of cancer of the colon, rectum as well as the stomach. It is reserved for prospective studies to estimate the valency of the determination of CEA or fetal sulphoglycoproteid antigen in the gastric juice for the recognition of early carcinomas of the stomach. At present the proof of an immune response of the tumor carrier to TAA in vitro seems to be more favourable as a second way of the immunological tumour diagnostics. Sensitivity as well as specificity might here at first higher be estimated, when they are theoretically established. Whereas the inhibition test of leucocyte migration and technically similar variants in apparently sufficient specifity have an insufficient sensitivity and thus give too many falsely negative results the mobility test of the electrophoresis of macrophages in the modification of the immunological tumour profile distinguishes itself by a high certainty and by evidence on the probable localisation of the organ and the primary tumour (up to now verified for the cancer of colon and rectum, carcinomas of the stomach). Apart from the research of further principles, however, even for this test technological simplifications are to be found, before a broader use in routine work becomes practicable.


Dependence of the antibody response of guinea-pigs to collagen on the type of adjuvant and on the conformation of the antigen. The immunological response to collagen of guinea-pigs is strongly dependent on the conformation of the antigen and on the type of adjuvant. Freund's complete adjuvant facilitated excellent delayed hypersensitivity skin reactions to native (triple helical conformation) as well as denatured (random coil conformation) collagen. Immunization of guinea-pigs with collagen in this adjuvant gave rise to very low levels of antibody to native collagen and failed to induce antibodies to denatured collagen. Use of Freund's incomplete adjuvant resulted in excellent antibody responses to native collagen, but it did not induce antibodies to denatured collagen. Animals injected with collagen and Freund's incomplete adjuvant were not sensitized for cell-mediated immunological reactions. The antibodies to collagen were specific with regard to collagen from various species but displayed different degrees of cross-reactivities depending on the species of collagen used for immunization. They were specific for the triple helical conformation of the collagen molecule.


[A vertebral body implant for use in palliative surgery of spinal tumors (author's transl)]. A newly developed vertebral body implant is discussed which is used to support and stabilize the spine after surgical removal of a vertebra in spinal tumors.


[Alpha 1 fetoproteins determination in the amniotic fluid -- a method for prenatal diagnosis]. It is reported on the evaluation of alpha1-fetoprotein in amniotic fluid by means of single radial immundiffusion method. Our results are compared with those in the literature. The importance of this method is discussed for antenatal diagnosis of spina bifida and anencephaly.


[Fetography and amniography in prenatal diagnosis of genetic disorders]. 68 patients were transfered to the prenatal diagnosis. Both the alpa-fetorotein-estimation and the fetography or amniography were performed in 13 patients. The indication for fetography is the exclusion of neural tube defects and other serious visible defects. The amniography can especially carried out in the prenatal diagnosis of defects of intestinal tract. A program is proposed to the prenatal detection of neural tube defects: 1. Examination of all of the pregnacies with ultra sonic B scan. 2. At suspicions ultra sonic B scan and after birth of a child with neural tube defect alpha-fetoproteinestimation at the 16th week of pregnancy and fetography at the 24th week of pregnancy.


Ultrastructural and cytochemical studies on anionic surface sites of Mycoplasma membranes. Negatively charged, chemical groups on the surfaces of mycoplasma membranes were studied in the electron microscope by staining with positively charged ferric oxide hydrosols in the acetic acid (AI-reagent) or propanoic acid (PI-reagent), respectively. The technique of thin sectioning was used. With the AI-reagent, no significant binding of the ferric oxide particles was observed on the mycoplasma membranes. With the lipophilic PI-reagent, the membrane surfaces of M. mycoides subsp. capri, M. mycoides subsp. mycoides, M. gallisepticum, and Acholeplasma laidlawii were heavily stained with focal aggregates of iron granules. M. hominis did not bind the label. Prior chemical and enzymatic treatments indicate that the cationic ferric oxide particles in propanoic acid interact with negatively charged, presumably lipid phosphate groups exposed on the surfaces of mycoplasma membranes.


[The influence of zink and lead on the formation of antibodies (author's transl)]. Female NMRI-mice were injected intraperitoneally and subcutaneously with zinc chloride and lead chloride as well as with a mixture of both. The mice were also immunized with foreign-proteins. To demonstrate the interconections, the heavy metals and the proteins were injected separately, as was the heavy-metal-protein-suspension. The results of the studies are as follows: 1. Intraperitoneal application of lead leads to a significant elevation of the antibody titer, using bovine immunglobuline while zinc applications showed no rise or slightly raised antibody formation. 2. It could be demonstrated that the admixtion of zinc chloride depresses the stimulatory effect of lead chloride on the immunization. 3. With the heavy metal-protein-mixture is proved that lead chloride stimulates the antibody production more than zinc chloride. To explain these results, it is discussed whether the adjuvant effect of lead is inhibited by zinc whereas zinc itself stimulates the immunsystem through another working mechanism.


Investigations on the carcinogenic burden by air pollution in man. XIV. Effects of automobile exhaust condensate on the Syrian golden hamster lung. Syrian golden hamsters were intratracheally instilled with 5 or 2.5 mg/animal of automobile exhaust condensate at two weekly intervals. Moribund animals were fixed by intravascular perfusion. Samples of lobar and segmental bronchi, as well as of peripheral lung tissue, were taken for electron microscopical examination. In addition, all organs were examined histologically. After a survival time of 30 to 60 weeks all animals developed multiple pulmonary adenomas, thus indicating a marked carcinogenic effect of automobile exhaust condensate.


[Possibilities of cervical fusion surgery in the spinal canal with intervertebral disk degeneration, Therapeutic and case-report contribution]. This is a report of a patient with pains in the shoulders and arms, atrophies of the small muscles of the hands, and paraspasms of the legs. After having suffered from the disease for eight to nine years, the patient was diagnosed as having cervical myelopathy. Measurements of the bony cervical vertebral column, which were made on a lateral roentgenogram, showed extreme narrowness of the spinal canal, the measured values obtained being below the lower limit. In addition, the cervical canal was narrowed at three levels by the formation of projections on degenerate disks. This suggested the most effective surgical treatment: Vetral clearance of disks, removal of prominences, and fusion of the three levels through the use of four bone dowels. Subsidence of pains in the shoulders and arms, improvements of paresis of the hands, and considerable improvement of paraspams of the legs were observed postoperatively. This particular case shows that surgical intervention may be considered a possible method of therapy even in the case of narrowness of the cervical canal, a condition that was aggravated here by a number of degenerate disks. In such situations it is possible for the cervical canal to be considerably relieved by clearing the disks, removing the projections and crests, and making use of the special technique of fusion or joining together so that improvement will be possible even in those cases where the disease has already arrived at a progressive state.


[Use of the indirect rat mast cell degranulation reaction in vitro for characterization of the specific activity of non-infectious allergens]. The test of indirect degranulation of rat mast cells was applied to the study of the specific activity of 16 series of noninfectious allergens from domestic dust (3 series) from hotel dust (7 series) and from the pollen of fescue (6 series). The concentration of protein nitrogen in the allergens from the domestic and hotel dust constituted to 10 000 PNU, and in the allergen from the pollen of fescue--125 000, 50 000, 15 000 and 7 000 PNU. In studying the specific activity of domestic allergens use was made of the sera of patients suffering from bronchial asthma with a marked allergy to domestic dust sera of patients with pollinoses with a marked allergy to fescue pollen was applied to the study of the specific activity of pollen allergen. For the assessment of the indirect degranulation test in mast cells of rats an index of mast cell degranulation of rats (IMCDR) was employed. It was shown that the IMCDR values were equal to 0.46, 0.58, 0.62 in using the domestic dust and in the case of the hotel dust - from 0.17 to 0.28. The mean values for the group of preparations from the domestic dust were 2.5 times greater than for the group of preparations for the hotel dust and were 0.55 and 0.22, respectively. For the allergen from the fescue pollen the IMCDR values varied from 1.23 to 0.16, and increased with the rise of the protein nitrogen concentration (PNU). Additional studies demonstrated that the specific activity of the domestic dust allergen with the crude sera and with those preserved in frozen condition for 3 to 6 months was almost the same. The results obtained permit to draw a conclusion that the method of indirect degranulation of rat mast cells with trizing of commercial batches of infectious allergens.


[Several allergic reactions under conditions of functioning of an inhibitor of antibody activity in the body]. Allergization under conditions of functioning an inhibitory factor in the organism was studied by the reaction of leukocytolysis with Tsuverkalov's dysenterin and in the reproduction of hypersensitivity of the immediate type on the Schultz-Dale's model. It was shown that in functioning an inhibitory factor in the organism the degree of leukocytolysis reaction with the dysentery allergen decreased in the immunized animals in parallel with the fall in the serum antibody activity. A depression of the activity of macromolecular sensitizing antibodies was also observed.


[Cerebral circulatory disorders in patients with cardiovascular diseases accompanied by cardiac rhythm disorders]. The authors examined 180 patients where disorders of cerebral circulation developed on the background of different disturbance of the heart rhythm. The patients suffered from rheumatic heart disease, hypertensive disease or atherosclerosis in concomitance with ischemic heart disease. For comparison, 100 patients with similar disorders were studied, but without disturbances of heart rhythm. It was established that disorders of cerebral circulation were more frequently seen in permanent forms of cardiac fibrillation and less so in paroxysmal cardiac fibrillation and extrasystoles. The most severe disorders of cerebral circulation were seen in combinations of permanent forms of cardiac fibrillation with single and frequent ventricular extrasystoles in patients with rheumatic heart disease.


[Experience with the etiotropic treatment of children with tick-bone encephalitis]. The authors conducted a comparative study of the therapeutical effectivity of ribonuclease, hetero- and homologic specific gamma-globulin used in 189 children with tick-borne encephalitis in the acute period of the disease. It was established that there was a significant advantage of ribonuclease before serotherapy. During ribonuclease treatment the febrile period of the disease was much less, while the meningeal symptoms disappeared more rapidly with normalization of the CSF.


Sideroblast score: A sensitive indicator of iron deficiency and hypoproliferative anemia. The detection and enumeration of sideroblasts depend critically on the method used for iron staining of bone marrow smears. Several methods proposed for semiquantitative evaluation of bone marrow hemosiderin (iron stores) were compared with respect to their suitability for detection of normal and abnormal sideroblasts. Instead of the customary percentage of sideroblasts, the introduction of a sideroblast score is proposed and its diagnostic relevance was prospectively studied. Low sideroblast scores are associated with iron deficiency and hypoproliferative anemia. A normal sideroblast score, despite the absence of stainable hemosiderin, exclused the diagnosis of severe iron depletion. Elevated sideroblast scores may be correlated either with iron overload and/or sideroblastic (sideroachrestic) anemias.


Familial thrombosis due to antithrombin III deficiency in a Greek family. A Greek family with hereditary antithrombin III (AT III) deficiency associated with venous thrombosis is reported. 5 members of the family were affected. In these patients, AT III and heparin cofactor activities were decreased. Immunoreactive AT III showed a positive correlation to both AT III and heparin cofactor activities. alpha2-Macroglobulin and alpha1-antitrypsin were normal. The pattern of inheritance of the defect is autosomal dominant.


[Development of tumors as the 1st manifestation of acute leukemia]. The acute Myeloblastosis in adults which forms tumours, is one of the very rare forms of this disease. The unusual development of the disease, the last appearance of characteristic symptoms and difficulties in forming a histological diagnosis usually makes it very difficult to diagnose this disease in sufficient time. On the basis of a 44 year old patient's course of disease, the clinical difficulties and the pathological-anatomical diagnosis of an acute Myeloblastosis that formed tumours, are discussed.


Effect of encephalitogenic protein on migration in agarose of leukocytes from patients with multiple sclerosis. Variable effect of the antigen in a large dose range, with a literature review. The literature on in vitro test of sensitivity to brain antigen in multiple sclerosis is summarized in the tables. A study is presented using leukocyte migration in agarose as test system and a wide range of concentration of bovine encephalitogenic protein as antigen. Definite reactivity was seen in 12 out of 28 patients with MS. Some occurred in patients who had had no signs of disease activity for at least 5 years. Reactivity occurred more often in patients with active disease studied within 5 days after a relapse. The significance of the in vitro reactivity is briefly discussed.


Effect of encephalitogenic protein on migration in agarose of leukoytes from patients with multiple sclerosis. A longitudinal study of patients with relapsing multiple sclerosis or with cerebral infarction. Ten patients with MS were studied with leukocyte migration in agarose technique to detect changes in reactivity ot encephalitogenic protein in connexion with a relapse. Six showed significant reactivity within a few days after the relapse. It decreased or disappeared during the 2 weeks after the relapse, but sometimes reappeared and was found in tests performed 2-3 months later. Five patients with cerebral infarction were studied in a similar way--in three, marked reactivity was noted within a few days after the stroke; in these, reactivity decreased or disappeared in later tests. In the two other patients, reactivity appeared in the second and/or third test. The possbility of a reactivity as an epiphenomenon due to CNS tissue destruction is discussed, and the need for antigens with a more restricted specificity for such an analysis is stressed.


Alpha-1 fetoprotein in cord serum correlated to gestational age. Measurement of alpha1-fetoprotein by rocket immuno-electrophoresis in cord serum from a consecutive series of 422 newborn infants showed a negative correlation with gestational age. The coefficient of correlation was divided by 0.70 for girls and divided by 0.74 for boys. For a given alpha1-fetoprotein concentration in cord blood the confidence limits for prediction of gestational age for both sexes were calculated to +/- 24 days. It is concluded that determination of alpha1-fetoprotein in cord serum is of limited help in the evaluation of maturity.


The influence of smoking on the haemostatic mechanism in pregnancy. A prospective study through pregnancy is described comparing smokers with non-smokers in an attempt to assess possible changes in the haemostatic mechanism. Various components of the coagulation and fibrinolytic enzyme systems were assayed serially up to 38 weeks of gestation. Plasma fibrinogen was lower in the smokers than in the non-smokers. Also plasminogen was slightly decreased and plasminogen activator activity and serum F.D.P. increased in the former group. Although the results failed to demonstrate motor disseminated intravascular coagulation in smokers, the pattern of a possible low grade syndrome emerged.


Effects of ophthalmic vehicles on the stability of the precorneal film. The break up time (B.U.T.) of the precorneal film has been studied before and after application of 34 different vehicles. The material examined comprised 646 eyes. Maximum increase of the B.U.T. was obtained with 2% methyl cellulose (four times) and 10% polyvinyl alchohol (seven times). These vehicles in the usually employed concentrations, fat-free ointment (polyethylene glycol), acetyl cysteine and polysaccharide (dextran) affected a less pronounced prolongation. The B.U.T. was reduced four or five times by fatty, anhydrous ointments and by silicone oil, about two or three times by emulsions and oils, and twice by 0.01% benzalkonium chloride. The clinical significance of the B.U.T. alterations is discussed.


K antigen determination of Escherichia coli by counter-current immunoelectrophoresis (CIE). The application of the counter-current immunoelectrophoresis (CIE) for determination of E. coli acidic polysaccharide K antigen is described. The most appropriate dilutions of antigens and antisera were established after examination of six different K antigens with homologous OK antisera. According to this result all test strains for acidic polysaccharide K antigens, i.e. K1 to K57, K62, K74, K82, K53, K84, K92, K93, K94, K95, K96, K97, K98 and K100, were examined and OK antisera suitable for the CIE test were selected. The following reciprocal cross-reactions were found: K2 and K62; K7 and K56; K12 and K82; K13, K20 and K23; K18 and K22; K16 and K97, and K53 and K93. The CIE method is now used as a routine test in our laboratory.


The patas monkey as a model for dihydrostreptomycin ototoxicity. Although the cochlear toxicity of dihydrostreptomycin (DHSM) is well-recognized in man, it has always proved difficult to demonstrate in animals. Hearing thresholds in M. nemestrina monkeys remained essentially unchanged after DHSM 100 mg/kg im daily for 8 months, but E. patas monkeys were severely deafened by DHSM 20 mg/kg for 90 days, a regimen formerly used in treating human tuberculosis. The patas monkey may prove to be the animal model of choice for evaluating aminoglycoside ototoxicity.


Cross-reactivity of Bacteroides fragilis O antigens. The cross-reactivity of O antigens prepared from Bacteroides fragilis, other Bacteroides species and from Fusobacterium has been examined by indirect haemagglutination and inhibition of haemagglutination. Fifteen of 20 B. fragilis ss. fragilis strains showed O-antigenic cross-reactivity with one or more of the test strains of B. fragilis ss. fragilis: NCTC 9343, Lille E 323 and SBL B55. The same applies also to 3 strains classified as B. coagulans, B. hypermegas and B. putredinis. The multispecificity of B. fragilis O antigens is pronounced. Test systems for demonstration of 9 specificities, all harboured by one or more of the 3 test strains, have been worked out.


Ultrastructure of the organ of Jacobson and comparative study with olfactory mucosa. The organ of Jacobson has been studied in lizards, rats, guinea pigs, rabbits and dogs by means of embedding in nitrocellulose, semithin sectioning, and electron microscopy. The various elements of the organ in the species studied have been analysed and a comparative study carried out between the sensory portion of the organ and the olfactory epithelium.


Protein concentration in interstitial and lymphatic fluids from the subcutaneous tissue. The protein content of interstitial fluid and lymph from the same tissue was determined fluid samples of nanolitre volumes being taken from the subcutaneous tissue of rabbits by micropuncture. In the same area lymph was collected from lymphatic vessels of 30-100 mum. An electrophoretic technique in polyacrylamide gels in glass capillaries was used for the analysis of proteins. The gels were stained and then scanned on a microscope equipped with a moving table and photomultiplier. The area under each protein band was calculated from the recorded densitograms. The method required only a few nanolitres of fluid for analysis. Using solutions of known protein concentrations the relationship between the amount of protein and the area under the densitogram band was investigated. This relationship was found to be linear, making it possible to quantify the protein content of both interstitial fluid and lymph samples. The interstitial fluid/plasma concentration ratios for albumin, transferrin, globulins and total proteins were 0.42, 0.42, 0.32 and 0.37. The albumin/globulin ratio in interstitial fluid was found to be 1.60 of the plasma value although the values obtained for lymph were similar to those found for interstitial fluid. The similarity between the protein concentration of interstitial fluid and lymph indicated that the endothelial lining of the terminal lymphatics did not restrict the passage of macromolecules into the lymphatics.


Carbonic anhydrase in the intestinal tract of the guinea-pig. The distribution of carbonic anhydrase activity in the intestinal tract of the guinea-pig was studied by the histochemical method of Hansson. Enzyme activity was demonstrated in epithelial cells, erythrocytes and capillary walls. In the gastric mucosa parietal cells, surface mucous cells and neck mucous cells were highly active. In the small intestine only a few epithelial cells on the villi and in the upper part of the crypts showed enzyme activity. They seemed to be randomly scattered among inactive ones. It is not clear at present if they represent a distinct cell type or specialized absorptive cells. In the proximal colon most surface epithelial cells were highly active (goblet cells were inactive), whereas the surface cells in the distal colon showed less activity with a more varying degree of staining. In the cecum enzyme activity was found in the surface epithelium and in the upper part of the crypts, the staining being most marked at the luminal border of the surface cells. The staining reaction was completely inhibited in all tissues by 10 muM acetazolamide, except for the luminal staining of the cecum, which was inhibited only by 100 muM acetazolamide. This indicates the presence of high concentrations of carbonic anhydrase, probably of the "low activity" form, at this locus. Mucosal scrapings were taken from the intestinal tissues, homogenized and assayed for carbonic anhydrase activity by a changing-pH indicator method. The results confirm those of previous studies and correlate well with the histochemical findings.


Possible linkage between group-specific component (Gc protein) and pure depressive disease. Genetic linkage was studied in pure depressive disease, a subgroup of unipolar depression defined by absence of familial alcoholism and/or antisocial personality. Rigorous research criteria were used for diagnosis and the diagnoses were made blind, i.e. without the knowledge of the genetic marker results. Under the assumptions of full penetrance and of all examined individuals' having passed the risk period, two out of eight families investigated showed a possibility of linkage, using the lod score method, with the Group-Specific Component (Gc protein) locus, one of the 17 polymorphic markers tested. The results, being based on a small amount of data, are presented as a hypothesis only; important possible implications of Psychiatry, however, indicate the desirability of testing the hypothesis.


Immunoglobulin receptors in lymphocytes. This discussion has been concerned with the immunoglobulin receptors of lymphocytes and particularly with the ability of these immunoglobulin receptors to bind specific antigens. We have considered some of the methods used to measure these antigen-binding cells (ABC) and the nature of the ABC in normal, nonimmunized animals. The response of the ABC to specific immunization has been discussed to demonstrate the usefulness of technics which allow the evaluation of precursor cell populations. Finally, the development of these ABC and more particularly the development of B lymphocytes in general has been considered and compared to the ontogenic pattern of several species.


Comparative effects of overdrive on sinus and subsidiary pacemaker function. Recent evidence has suggested a difference in response to overdrive pacing dependent on the location of the pacemaker within the A-V conduction system. To test this hypothesis, the effects of overdrive pacing were evaluated in five anesthetized dogs with experimentally induced A-V block and in seven patients with advanced A-V block. In the animals, sinoatrial node recovery times were studied over wide ranges of rates (130 to 210 beats per minute) and durations (30 to 180 seconds) of atrial pacing. All sinus node recovery times were less than 600 msec. with a mean maximum pause of 0.540+/-0.043 seconds (M.+/-S.E.M.). In contrast, after ventricular pacing (rates 90 to 150 beats per minute; durations 30 to 180 seconds), subsidiary pacemaker recovery times were significantly greater (p less than 0.025) with a mean maximum recovery time of 28.4+/-8.3 seconds. In the seven patients studied, all sinus node recovery times were less than 1,400 msecs. with a mean maximum pause of 0.954+/-0.051 seconds. As seen with the experimental animals, a significantly longer (p less than 0.025 mean maximum subsidiary pacemaker recovery time of 3.55+/-0.92 seconds was observed. The present studies in both experimental animals and in man without evidence of sinus node dysfunction showed that sinus node recovery time was independent of both rate and duration of atrial overdrive pacing. In contrast, subsidiary pacemaker recovery time was correlated with both rate and duration of ventricular overdrive pacing. In both experimental protocols, subsidiary pacemaker recovery time was shown to exceed sinus node recovery time at all rates and at all durations of pacing. Postrecovery sinus node acceleration was consistently observed after atrial overdrive pacing. In contrast, postrecovery subsidiary pacemaker "depression" characterized ventricular overdrive pacing. It is concluded that subsidiary pacemakers are significantly more susceptible to overdrive suppression than the sinoatrial node, a feature of substantial clinical significance.


Arrhythmias documented by 24 hour continuous electrocardiographic monitoring in 50 male medical students without apparent heart disease. Results are reported of portable 24 hour dynamic electrocardiographic monitoring in 50 male medical students without cardiovascular disease, as defined by normal clinical and noninvasive cardiovascular examination. During waking periods, maximal sinus rates ranged from 107 to 180 beats/min (mean +/- 5). Twenty-five subjects (50 percent) had episodes of marked sinus arrhythmia as defined by spontaneous changes in adjacent cycle lengths of 100 percent or more. Fourteen subjects (28 percent) had sinus pauses of more than 1.75 seconds, usually during sinus arrhythmia. Transient nocturnal type I second degree atrioventricular (A-V) block was noted in three subjects (6 percent). Of 28 patients (56 percent) having atrial premature beats, only 1 (2 percent) had more than 100 such beats (141) in 24 hours. Of 25 patients (50 percent) having premature ventricular contractions, only 1 (2 percent) had more than 50 such contractions (86) in 24 hours. In conclusion, frequent atrial and ventricular premature beats are unusual in a young adult male population. In contrast, bradyarrhythmias (including marked sinus arrhythmia with sinus pauses, sinus bradycardia and nocturnal A-V block) are common. These findings are useful in evaluating the clinical significance of arrhythmias detected with portable monitoring.


A fixative for intestinal parasites permitting the use of concentration and permanent staining procedures. Results obtained with stools preserved in a simple, stable and relatively nontoxic fixative (SAF) composed of sodium acetate, acetic acid and formalin, suggest that this fixative is a suitable alternative to other fixatives for the preservation and recovery of intestinal parasites by diphasic concentration methods and permanent staining.


Mitral valve prolapse: a review of associated arrhythmias. The syndrome of mitral valve prolapse with associated auscultatory-electrocardiographic findings is now well documented. Two representative cases of repetitive tachyarrhythmias in patients with mitral valve prolapse are discussed as well as an analysis of the 589 cases in the English literature of arrhythmias in patients with mitral valve prolapse. The average age of the patients was 38 years; 70 per cent of them were women. Symptoms were variable, but palpitations occurred in 44 per cent, lightheadedness in 12 per cent and syncope in 4 per cent. Premature atrial and/or ventricular contractions were found in 55 per cent, premature ventricular contractions in 45 per cent, supraventricular tachycardia in 6.1 per cent and ventricular tachycardia in 6.3 per cent. Sudden death was noted in 1.4 per cent. A discussion of the pathogenesis of arrhythmias and therapy concludes this review.


Disseminated intravascular coagulation induced by progesterone in the pregnant rat. Prevention by estogens. Fatal disseminated intravascular coagulation (DIC) was induced in female rats by administration of progesterone in late pregnacy. This prevented parturition, with intrauterine fetal death 2 to 4 days past term and subsequent retention of dead fetuses. Concomitantly with or closely following the intrauterine death of their litters, a large proportion of pregnant rats died with histologically evident DIC. Administration of cortisone, heparin, or disoumarin did not prevent DIC, and xi-aminocaproic acid, acetylsalicylic acid, or an onion-rich diet tended to increase its incidence. Antibiotic regimens gave variable results, with significant decrease in DIC only with a combination of two wide-spectrum penicillins. Neomycin and polymyxin had little effect on susceptible Sprague-dawley derived rats, but polymxin caused a significant increase in DIC in a resistant strain of hooded rats. Fatal maternal DIC was completely prevented only by use of natural or synthetic estrogens concurrently with progesterone, although this did not alter the sequence of abnormally prolonged pregnacy with intrauterine fetal death and retention of dead fetuses. Potencies of estrogens varied greatly, but all compounds tested prevented DIC at adequate dosage levels. Diethylstilbestrol, the most potent drug tested, was completely protective at 1 mug daily given subcutaneously. beta-Estradiol was the most effective natural estrogen, giving complete protection with a 10-mug daily subcutaneous injection. Estrogens were much more potent by subcutaneous injection than by oral ingestion, and toxic side effects were sometimes noted with higher levels of the latter. For estrogen therapy to be effective, it was necessary to begin its use before the expected onset of DIC, and in no instance was there evidence of reversal of this process after signs of illness were observed.


Molecular interactions in human atherosclerotic plaques. Most plasma proteins appear to be present in intima at concentrations that are a linear function of molecular weight and concentration in the plasma. Thus low density lipoprotein (LDL) (molecular weight, 2 X 10(6)) has the greatest retention relative to its plasma concentration, whereas the relative retention of albumin is only 15% of the relative retention of LDL. This gives rise to the concept that "whole plasma" crosses endothelium, and the steady state concentrations reflect rates of egress of the macromolecules, which in turn depend on molecular sieving. Fibrinogen is a major plasma protein in intima in addition to LDL and albumin, and there are also substantial amounts of the protease inhibitors alpha2-macroglobulin and alpha1-antitrypsin. Intima also contains insoluble derivatives of plasma--extracellular cholesterol, both free and esterified, and fibrin. The balances of intact LDL/"deposited" cholesterol and of fibrinogen/fibrin are closely linked with intimal morphology. Fibrinogen and electrophoretically mobile LDL are increased about threefold in gelatinous lesions, whereas there are only slight rises in fibrin and deposited cholesterol. In the deep layers of fibrous plaques, fibrin is increased fivefold and cholesterol up to thirtyfold. In these lipid-rich layers, LDL is rapidly lost on incubation of tissue samples, but in some gelatinous lesions it first increases and only decreases on longer incubation, suggesting release of a previously immobilized lipoprotein fraction. This immobilized lipoprotein was investigated by subjecting tissue samples to immunoelectrophoresis to remove mobile LDL and tissue enzymes, followed by treatment of the tissue with enzyme and measurement of the lipoprotein released on fresh immunoelectrophoresis plates. Plasmin or a crude collagenase released large amounts of lipoprotein from samples of amorphous atheroma lipid. For all samples the amount of lipoprotein released was highly correlated with the accumulation of deposited cholesterol, suggesting that immobilization of LDL may be an intermediate step in the irreversible deposition of extracellular cholesterol. Plasmin is highly effective in releasing immobilized lipoprotein, and the concentration of immobilized lipoprotein is significantly correlated with the concentration of insoluble fibrin, suggesting that the lipoprotein may in some way be immobilized by fibrin.


Further evidence for a central regulation of free fatty acid mobilization in the rat. Previous studies have shown that anterior hypothalamic deafferentation in rats completely suppresses the increase in plasma free fatty acid (FFA), but not the hyperglycemia induced by administration of 2-deoxyglucose, suggesting a specific central regulation of FFA mobilization. The physiological importance of this finding was further investigated by examining in deafferented rats the response to several stimuli that modify the rate of lipomobilization in normal rats. The results show that the hypothalamic lesion interferes with FFA mobilization mainly when increased sympathetic activity is required: during cold exposure or forced muscular activity, and after insulin-induced hypoglycemia or a relatively long period of fasting. Changes in blood sugar responses, when observed, could be interpreted as secondary to an initial block in FFA mobilization. The data support our hypothesis that there are areas in the central nervous system sensitive to glycopenia and activated in situations requiring rapid mobilization of metabolic reserves that can specifically influence FFA mobilization through an activation of the sympathetic fibers of adipose tissue.


Arotic Evans blue dye accumulation: its measurement and interpretation. Parenteral Evans blue dye (EBD) binds to albumin and thus can indicate transvascular protein flux. A model was developed to relate the arterial surface accumulation of EBD to the light refected from the opened vessel surface. The accumulation, M (nmol cm-2), was related to the optical "absorbance" (rho) of the surface by the equation, M = 3.92 rho + 0.80 rho3, where rho = -ln(I/I0) and I and I0 are the intensities of light reflected from the surface in the stained and unstained state, respectively. The validity of the model was examined in two ways, 1) by comparing the value of the optical absorption coefficient for EBD predicted by the model to the measured value: the predicted value was 0.128 +/- SD 0.015 cm2 nmol-1, the measured value was 0.125 +/- SD 0.005 cm2 nmol-1; and 2) by comparing predicted changes in vessel wall stretch to measured values: the percentage changes (Y) in area predicted by the model from changes in M were linearly related to the directly measured percentage area changes (X) by Y = 0.09 + 1.01X, +/- SD 2.54, with a correlation coefficient of 0.994.


Electroimmunoassay of alpha-2-opsonic protein during reticuloendothelial blockade. Physiological regulation of reticuloendothelial (RE) phagocytic activity by a plasma opsonic factor has been documented. In the recent study, serum levels of this alpha-2-opsonic protein in rats during colloid-induced RE blockade were measured utilizing an electroimmunoassay (Rocket immunoelectrophoresis) with monospecific antiserum to the purified alpha-2-glycoprotein. RE blockade was produced by the intravenous injection of the gelatinized "RE-test-lipid emulsion" at a dose of 50 mg/100 g body wt. The opsonic activity of serum at various intervals during colloid-induced RE blockade as measured by tissue slice bioassay manifested a high correlation (r = 0.98) with the serum opsonic protein concentration as measured by the electroimmunoassay. During RE blockade (30 min), there was a rapid depletion of the opsonic alpha-2-glycoprotein to 20% of the initial preinjection levels. Serum concentration of this glycoprotein remained low for at least 2-3 h after which time its concentration progressively increased with approximation of normal values by 6 h postblockade. Opsonic protein concentration at 24 h postinjection were significantly (P less than 0.05) elevated above controls. Thus, colloid-induced RE blockade is associated with the removal of this glycoprotein from the serum and recovery from RE blockade is accompanied by a restoration of opsonin levels. The electroimmunoassay can provide a sensitive technique to monitor this humoral factor known to exert a physiological control on the RE system.


A complement-fixing antigen from Trypanosoma cruzi grown in cell cultures. A complement-fixing (CF) antigen was prepared from amastigotes and trypomastigotes of Trypanosoma cruzi (Ernestina strain) grown in beef embryo cell cultures. Multiple lots of the antigen, which consisted of a supernate of washed and disrupted organisms, required material from 10(6) to 10(7) total organism per ml for optimum CF activity. Antibody at dilutions up to 1:256 was demonstrable in various sera from infected animals or patients. Contaminating beef cells from infected cultures were shown to be partly responsible for crossreactions of the antigen by CF with sera from cases of cutaneous leishmaniasis in whom concomitant infection with T. cruzi could be excluded. There were no cross-reactions with syphilitic sera and the frequency of positive reactions with normal sera was very low. Some characterisitics of the antigen included stability to storage at -20 degrees C and -70 degrees C for months, inactivation at 60 degrees C and by lyophilization, and an estimated molecular size of between 50,000 and 100,000 on the basis of membrane filtration.


Comparison of cell culture with epimastigote antigens of Trypanosoma cruzi. Various epimastigote antigens of Trypanosoma cruzi were compared with an amastigote-trypomastigote antigen from infected cell cultures by complement fixation (CF) and gel diffusion (Ouchterlony). CF results with human Chagasic sera showed that the amastigote-trypomastigote antigen was usually more sensitive than epimastigote antigens tested. In addition, cross-reactions with normal and other sera were no greater and perhaps less frequent than with crude epimastigote antigens. Specificity of the amastigote-trypomastigote antigen, however, was less than with a protein extract of epimastigotes. Gel diffusion results with human Chagasic and hyper-immunized rabbit sera indicated differences between epimastigote and amastigote-trypomastigote antigens whereas differences by CF with the same sera were equivocal.


Experimental gerontology and quantitative cytology: a new approach to cellular ageing. The application of the followiny cytologic methods for gerontological basis research is discussed: 1. Quantitative cytomorphology including electron microscopic morphometry and its combination with cytochemical reactions. 2. Histochemical studies on cell components like DNA in situ by means of acridine orange microfluorimetry and thermal denaturation. 3. X-ray microanalysis for measuring the intranuclear electrolyte contents. The results obtained indicate the usefulness and the validity of a quantitative cytological approach to the cellular ageing.


[Effects of premedication, narcosis and surgery on the coagulation and fibrinolysis systems and the platelets. The influence of dextran and hydroxyethyl starch (hes) during and after operation (author's transl)]. Patients who had undergone orthopaedic surgery were investigated. The changes attributable to premedication and narcosis were characterized by a primary fibrinolysis which was accompanied by a slight hypercoagulability. This increased fibrinolytic activity was more pronounced immediately after the beginning of the operation. The post operative changes are characterized by increased ADP-induced aggregation, increased release of platelet factors 3 and 4 and hypercoagulability with reduced fibrinolysis. The reduction in platelets during the operation could be prevented due to the influence of dextran and hydroxyethyl starch (HES). It came further to a slight increase in the activity of factor VII, to an increased fibrinogen polymerization and also to an increased release of platelet factor 4.


The effect of endotoxin on the mast cell c'AMP system. To determine whether the endotoxin induced release of histamine is mediated via the mast cell c'AMP system, hamster mast cells were isolated and incubated (prior to endotoxin-serum stimulates (ET-S) with disodium cromoglycate, isoproterenol and aminophylline. All drugs caused significant inhibition of the ET-S histamine release. The authors conclude that ET-S utilizes the c'AMP system to release histamine.


Immune and biochemical mechanisms in the allergic disease of the upper respiratory tract; role of antibodies, target cells, mediators and eosinophils. The pathways leading to the development of the allergic state and subsequently to the characteristic inflammatory response are complex in nature and result from an interplay between immunologic and biochemical events. Along these pathways a number of intrinsic factors, i.e., handling of antigens at mucosal level, transient immunodeficiency states, especially in the secretory IgA system, impairment in the IgE regulatory mechanism, modulation of cyclic nucleotides leading to mediator release and a "feedback" inhibition control provided by histamine and eosinophil derived products greatly dictate the outcome of events associated with allergic inflammation.


Total serum protein, serum protein fractions and serum immunoglobulins in colostrum-fed and colostrum-deprived calves. Total serum protein levels, serum protein fraction levels, and specific serum immunoglobulin class or subclass levels were measured in colostrum-fed (CF) and colostrum-deprived (CD) calves during the first 144 hours after birth. Total serum protein values increased at 24 hours in the CF group and then decreased slightly at 144 hours. The increase in total serum protei5) in beta1-, beta2-, and gamma-globulins. The beta2- and gamma-globulin levels decreased by 144 hours, while the serum level of beta1-globulin continued to increase. The CD calves exhibited a significant decrease (P less than 0.05) in total serum protein at 24 hours, folhours, the level of beta1-globulin inlowed by a significant increase (P less than 0.05) at 144 hours. At 24 hours, the level of beta1-globulin decreased slightly, and the level of beta2- and gamma-globulins increased slightly. At 144 creased, and the level of gamma-globulin decreased. The beta2-globulin level did not change. At birth, immunoglobulin (Ig) M was detected in 5 of the 10 calves, IgG1 in 6 of the 10 calves, and IgG2 in 3 of the 10 calves. By 24 hours after birth, all CF calves had detectable levels of IgM, IgG1, and IgG2, and there were significant increases (P less than 0.01) in the mean serum levels of all 3 immunoglobulins. By 144 hours after birth, the serum levels of IgM, IgG1, and IgG2 decreased to various degrees. At 24 hours, the IgM level had not increased in CD calves; however, the level of IgG2 appeared to increase slightly, and the mean IgG1 level increased by approximately 50%. By 144 hours after birth, there was a significant increase (P less than 0.01) in the mean level of serum IgM. The level of IgG, also appeared to increase substantially, while the level of IgG2 appeared to increase slightly.


[Use of two dimensional immunoelectrophoretic method in the study of the antigenic relationships of "Mycobacterium simiae" and "M. habana" (author's transl)]. Following the cultural, biochemical and serologic studies reported elsewhere, it was thought of interest to investigate the antigenic structure of these strains. In the present work, sonicated suspensions of the bacteria were used as antigens, together with rabbit immune serum antibodies. The optimal conditions to obtain the largest number and the sharpest lines possible, were established. In this manner, we were able to obtain on two dimensional electrophoresis patterns were obtained. Using an anti-M. habana 4238 antiserum, 36 lines of precipitation were obtained with M. habana 4238, 41 lines with M. simiae 29 and 29 lines with for M. simiae 59-IX-7. Subsequently, identity tests were performed to verify the occurrence of were common antigens. These tests revealed such commons antigens in all the three strains. The comparison between M. habana and M. simiae 29 showed the occurence of at least one Precipitation line, that is characteristic of M. habana. These results appear to agree with those of Meissner, who obtained a small amount of anti-M. habana specific agglutinins. However, this worker did not believe that this was sufficient evidence to separate M. simiae 29 from M. habana. The antigenic differences between M. habana and M. simiae 59-IX-7 are more significant and appear to justify their differentiation into 2 serotypes: M. habana 4238 and M. simiae 29 being of serotype 1, M. simiae 59-IX-7 of serotype 2. The difficulties experienced in the separation of the antigenic fractions in the immunoelectrophoretic diagrams, led to consider the purification of the antigens and then to attempt to isolate the specific antigenic fractions.


[Paraneoplastic ichthyosis. Ultrastructural study]. A case of ichthyosis in association with bronchial carcinoma is studied by electron microscopy. Keratohyaline granules are always present in only one layer; variable in size but often reduced upon short tonofilament bundles, they are structurally normal. This suggested a reduced synthesis of a normal keratohyalin, different from the synthesis of an abnormal few spongy keratohyaline in autosomal dominant ichthyosis vulgaris.


An improved method for processing early rat otocysts for the electron microscope. Various methods of dissection, fixation, osmication, sectioning and staining were tested in order to develop an acceptable technique for preparing 9, 10, 11, and 12-day-old rat otocysts for electron microscopic study. The general problems associated with embryonic tissue-difficult handling, high water content and poor stainability are discussed, and concrete methods of preparation which significantly decrease these difficulties are proposed. The specific fixation and sectioning requirements of rat otocyst are also described in the elaboration of a method which will be used in subsequent studies of the organogenesis of rodent ear.


The inhibition of arginase from the hepatopancreas of a terrestrial snail by amino acids. The effects of L-amino acids on arginase from the hepatopancreas of the snail Ariophanta (=Cryptozona) ligulata were studied. This enzyme was inhibited by ornithine, valine, lysine, leucine, isoleucine, proline and threonine. The other amino acids were without any significant effect. Only ornithine was a non-competitive inhibitor, where as all the other inhibitory amino acids were competitive.


[Radiometric detection of formate 14C and formaldehyde 14C oxidation in folic acid deficiency]. An ionization chamber method was used in vivo to demonstrate a delayed oxidation of [14C] formaldehyde and [14C] formate to 14CO2 in folic acid-deficient rats as compared to control rats or folic-acid-deficient rats treated by folic acid. Results obtained showed that oxidation of these two molecules required the presence of folic acid.


[Comparison of the compliance of resting and contracting muscle]. Frog sartorius muscles are stretched at rest and during maximal tetanic contractions. Parallel compliance decreases when the length increases. The relationship between compliance and length is linear in double-logarithmic scale. The compliance of the active muscle (tetanic contraction) is not related to the length. The series-compliance is calculated from the parallel compliance and the active one. It increases with the length of muscle. These results are discussed on the basis of the sliding-filaments theory.


Response of isolated axonal preparations to hyperosmoticity: evidence for a cellular volume regulation. When submitted to a hyperosmotic stress, isolated axons of Callinectes sapidus shrink. Volume regulation is observed if the experiment is carried out by using serum from a crab adapted to sea water. 3' : 5'-AMP and dibutyryl AMP are also effective. A compound isolated from the hemolymph and having a molecular weight close to 10 000 mimics the effects of the serum and induces a partial regulation of the axonal volume.


[Immunological importance of a new protein: the beta2-microglobulin (author's transl)]. The Authors are presenting a concise review on a protein, beta2-microglobulin, that has gained in notable importance in the immunological field during these last years. The story of this protein begins with nephrology because it has a notable importance in the diagnosis of renal tubular diseases; in fact its presence in the electrophoretic pattern of proteinuria puts in evidence renal tubular damage. Nextly immunological studies have shown the beta2-microglobulin has strict correlations with the HL-A system; probably it forms a part of this system, as shown by the new immunofluorescent methods applied to the lymphocte. A recent study has shown also the beta2-microglobulin serum levels are very high in some malignant diseases, like in acute leukemia; however this low molecular weight protein could also be the expression of malignancy because it can be a product of non mature cells in a large quantity.


[Problems of the prenatal prophylaxis of the viral infections (author's transl)]. The present review will supply an simple notice over the characters of every one viral infection interesting the materne-faetal conditions, and also propose the most convenient intervention for each observed type. A not small lot of these problems are not resolved in the field of the antiviral immunity regarding the mother and newborn.


Effects of nonsteroidal anti-inflammatory drugs on the uptake of various cations by lymphoid cells. Several acidic nonsteroidal anti-inflammatory drugs (NSAID) as well as their corresponding alcohol molecules which are known to induce swelling of isolated lymphocytes by changing cell membrane permeability to water, are demonstrated also to induce changes of membrane permeability of lymphoid cells to one divalent cation, calcium, and to three monovalent cations, rubidium, cesium and sodium. According to the cells ionic environment, they increase or decrease the cellular uptake of cation which is itself also closely dependent on the ionic composition of the incubation medium. This drug-effect is very rapid, directly related to the medium NSAID concentration and almost totally reversible except to the most potent drugs such as flufenamic acid. Changes in intracellular ionic balance could have important catalytic effects on the metabolism of normal as well as of pathological cells. This fact could explain side-effects of these drugs as well as some of their therapeutic effects.


Anaerobic metabolism of the common cockle, Cardium edule. II. Partial purification and properties of lactate dehydrogenase and octopine dehydrogenase. A comparative study. 1. Octopine dehydrogenase and lactate dehydrogenase were purified 190-fold and 10-fold respectively from the adductor muscle of the marine bivalve Cardium edule by gel filtration on Sephadex G-100 and chromatography on DEAE-Sephadex A-50. 2. Lactate dehydrogenase was capable to convert D- and L-lactate, had a molecular weight of about 70 000 and 280 000 daltons, exhibits no distinct pH optimum and was not inhibited by lactate. The enzyme showed apparent Km values of 0.16 mM for pyruvate and 16 mM and 48 mM for D- and L-lactate respectively. 3. In comparison to the purified enzymes from other species, octopine dehydrogenase from Cardium edule showed similar biochemical properties : pH optima of 6.8 and 8.7 respectively, Km values of 0.9 mM (for pyruvate) and 2.0 mM (for arginine), a molecular weight of 37 000 daltons and inhibition by octopine. Electrophoretic studies on standard polyacrylamide gels showed five isoenzymes. 4. The biochemical properties of both dehydrogenases are compared to the conditions in vivo of these animals and the biological role of the octopine dehydrogenase is discussed.


Effect of vitamin B6 on gamma-aminobutyric acid level in rat brain. Vitamin B6 injected intraperitoneally into rats 400 mg/kg body weight, has produced a statistically significant decrease in GABA level concentrations in hippocampus and cerebellum. Cerebral cortex, caudate nucleus and thalamus have shown the decrease in GABA concentrations, but these changes were not statistically significant. No remarkable behavioural changes were noted under such circumstances. The possible functional meaning of these results is discussed in relation to the role of GABA distribution in different brain regions and development of convulsions.


Incorporation of double-labelled valine into delta-(L-alpha-aminoadipyl)-L-cysteinyl-D-valine by Penicillium chrysogenum. The incorporation of valine into the LLD-tripeptide, delta-(L-alpha-aminoadipyl)-L-cysteinyl-D-valine, a precursor of penicillin, was studied by incubating mycelial mats of Penicillium chrysogenum Wis. 49-2105 with double labelled valines. L-valine was incorporated into the LLD-tripeptide without formation of an alpha, beta-didehydrovaline intermediate. Intact D-valine was not incorporated into the LLD-tripeptide.


Absence of a prostaglandinic mediator in the lateral hypothalamic cardio-vascular inhibition. The cardio-vascular inhibition elicited by electrical stimulation of the paraventricular nucleus in the lateral hypothalamus of anaesthesized and desafferentiated dogs is not linked with a mediator release like PGE. The effect of this biological agent is not registered in the isolated femoral artery of a receiver, after deviation by a cruised circulation of the arterial blood of a hypothalamic stimulated donor dog. Antipyretics, which are also prostaglandin-synthetase inhibitors, enhance the lateral hypothalamic reactions. As the thermoregulation centre is localized in the same region as the cardio-vascular inhibition centre and because lowering of temperature depends upon vasodilation and decrease in the general cellular metabolism, both functions of the paraventricular nucleus activity, a hypothesis is proposed that thermoregulation and cardio-vascular inhibition centres are a functional and anatomical unity.


The effect of thermic stress on the somatic reaction of rage and on rapid circling turns, in the cat. An investigation was made of the effect of thermic stress on the somatic rage reaction and on rapid circling turns in cats awake and free to move in a behavioural cage. An increase in room temperature had a two-phase effect on the excitability of the nervous structures stimulated that is able to evoke the somatic rage reaction and rapid circling turns. The first phase, at room temperature 25 degrees-30 degrees C, was characterized by hypoexcitability; the second phase, appearing after longer periods of exposure and at temperatures above 30 degrees C, was characterized by the onset panting, hyperexcitability of the nervous structures stimulated and then by lowering of the somatic rage reaction threshold and a very significant increase in the number of circling turns. Experimentation on the action of cold external temperature on the excitability of structures involved in evoking the somatic rage reaction and rapid circling turns indicated an increase in their excitability, corresponding to an increase in spontaneous and evoked somatic motor activity.