[Possibilities of the quantitative structural analysis of prostate biopsies]. Stereological methods allow the determination of volumes, surfaces or number of tissue and/or cell compartments. The quantitative data on human biopsy specimens of the prostate suggest a diminution of the secretory activity of the glandular cells, while the smooth muscle cells are activated: the relative volumes of the Golgi areas and the rough endoplasmic reticulum are increased. Stereology will be a valuable tool for objective search on hormonal dependence of the glandular as well as the smooth muscular cells. Therein, the role of the smooth muscle cells in benigh prostatic hyperplasia should be focused.


Effect of temperature on rate of goldfish optic nerve regeneration: a radioautographic and behavioral study. Optic nerve regeneration was examined in goldfish radioautographically, as well as by a variety of behavioral measures of visual function. The rate of regeneration was markedly enhanced by increasing temperature above the ambient level (20 degrees C). The earliest functional indication of recovery was the appearance of an autonomic response to a change in illumination, then the ability to localize food visually and at a still later time, an optomotor response. Even the early behavioral indications of return of function appeared some time after there was radioautographic evidence that the tectum had been reinnervated.


Bilateral tectal innervation by regenerating optic nerve fibers in goldfish: a radioautographic, electrophysiological and behavioral study. Following unilateral enucleation and optic nerve crush in goldfish, the remaining nerve regenerates and innervates both optic tecta. Approximately 5% of the nerve fibers reach the ipsilateral optic tectum (IOT) via the ipsilateral tract at the chiasma. Comparable debris in both tracts was not sufficient to result in an IOT projection since when both nerves were crushed simultaneously the usual pattern was seen, i.e., each nerve innervated a contralateral optic tectum (COT). When the arrival of one nerve at the chiasma was delayed by staggering the nerve crushes, the nerve that first arrived at the chiasma partially innervated the Iot. In most instances the entire IOT was innervated, however, the stratigraphic distribution of fibers in the various tectal lamina was atypical. Electrophysiological analysis indicated that fibers from each area of the retina innervated the IOT visuotopically. The COT was ablated in order to determine whether the IOT projection could mediate behavior. All fish failed to respond to changes in illumination as measured by respiration and failed to swim with or against the stripes in an optomotor drum. Thus, the IOT input, possibly because of its sparseness, could not be shown to be behaviorally functional.


The metabolism of nerve terminal glycoproteins in the rat brain. The fate of [3H]fucose in subcellular fractions has been followed for a period of three weeks after intraventricular injection in the rat brain. Isolated synaptosomal membranes were examined by gel electrophoresis. Seven fucosyl glycoproteins with molecular weights ranging from 123,000 to 31,000 were detected by scintillation counting of transversely sliced gels and their specific activities determined. The half-lives of 6 of the membrane glycoproteins fell within the range 11-22 days. The smallest glycoprotein showed anomalous behaviour; its radioactivity did not fall significantly over the period of study. The results are discussed in relation to current concepts of fast axonal transport.


Identification of cells of origin of tectopontine fibers in the cat superior colliculus: an experimental study with the horseradish peroxidase method. The pontine projections from the superior colliculus in the cat have been studied by means of retrograde axonal transport of horseradish peroxidase (HRP). Following injections of HRP in the dorsolateral pontine nucleus, where the tectopontine fibers terminate, a fair number of labeled cells are found throughout the rostrocaudal extent of the ipsilateral superior colliculus. Relatively few of the labeled cells are of medium size (25-40 micron in diameter), more than 80% are small (10-25 micron), but no large cells are labeled. The cell bodies giving rise to tectopontine fibers are distributed in tectal layers deeper than the optic stratum (including this), with only a few in the deeper portion of the superficial gray layer. There are only few labelled cells in the relatively large lateral portion of the intermediate and deep gray layers were the largest neurons (more than 40 micron) are located. Most of these presumably belong to the tectoreticular and the tectospinal projections. The tectal neurons, distributed in various collicular layers, are supposed to receive different kinds of information from other parts of the central nervous system, e.g. from the retina, the cerebral cortex, the brain stem reticular formation, the spinal cord etc. The dorsolateral pontine nucleus appears to have a particular function in the integration of the input from the superior colliculus with those from other sources, especially from the inferior colliculus and the auditory cerebral cortex.


A methodology for the production of carbohydrate-specific antibody. 8-Methoxycarbonyloctyl glycosides are recommended as precursors for the preparation of artificial antigens that possess strongly immunodominant carbohydrate antigenic determinants. The preparation and immunochemical properties of a selected number of these antigens are reported to demonstrate the high levels of hapten-specific antibodies that are normally achieved using standard immunization schedules. The method, based on glycoside synthesis, promises to make available a wide spectrum of well defined antigens and the homologous antibodies.


alpha-Fetoprotein production and erythropoiesis in cell cultures of human fetal liver. alpha-Fetoprotein and the synthesis of heme associated with hemoglobin were measured simultaneously in short-term cultures of human fetal liver cells to correlate the relationship of alpha-fetoprotein to erythroid cell function. Both synthetic processes decreased exponentially during the first 5 days of culture. The use of media supplemented with different batches of fetal calf serum and porcine portal vein serum indicated that the optimal conditions for the production of alpha-fetoprotein were different from those required for the synthesis of heme associated with hemoglobin. Moreover, the alpha-fetoprotein-producing cells could be separated from erythroid cells after velocity sedimentation in Ficoll gradients. Although it is well known that erythropoiesis and alpha-fetoprotein production occur simultaneously during ontogenesis, alpha-fetoprotein itself (0.01-100 micron g/ml) did not stimulate heme synthesis in liver erythroid cells. Erythropoietin did not stimulate alpha-fetoprotein production. It is concluded that there is no cause-effect relationship between alpha-fetoprotein production and erythroid cell fuction in human fetal liver cells and that the two processes occur independently in different cell types.


Production of slow alpha2-globulin in the pregnant rat. Slow alpha2-globulin (salpha2G), a high molecular weight glycoprotein, appears as a component of rat serum in a wide variety of both physiologic and pathologic conditions, including pregnancy; it is also present in the neonate. In the present study, the protein was detectable in serum after 6 to 10 days, and was present at moderate levels on day 13, and thereafter through the rest of pregnancy. From immunofluorescent localization and 14C-labelled amino acid incorporation studies, salpha2G was found to be localized in uterine and placental tissues and to be synthesized by these tissues as early as day 6 of gestation. Production continued throughout pregnancy. Synthesis in the liver of the pregnant rat began at day 17 of gestation, which is in contrast with the observation in pathologic conditions that liver synthesis is an initial source of the serum protein; substantial fetal liver synthesis was occurring at day 21 of gestation, and the amount in fetal serum was four times greater than that in maternal serum at that time. It is likely that the fetal liver produces salpha2G as soon as it begins to function.


Myocardial infarction size: measurement and modification. The majority of in-hospital deaths from acute myocardial infarction occur as a result of the "power failure" syndrome (severe congestive heart failure and cardiogenic shock), which results from extensive loss of myocardium. The death of myocardial cells is sequential over many hours. Surrounding the central zone of necrosis in an acute myocardial infarction is a zone of ischemic myocardium whose fate might be altered by interventions during the early phase of the infarction. ST-segment mapping, serial measurement of the serum concentration of creatine phosphokinase and myocardial imaging by means of radionuclides are being developed for the noninvasive assessment of infarct size in animals and humans. A number of interventions appear to limit infarct size in animals. There have been relatively few studies in humans to date, but preliminary results suggest that infarct size might be limited by certain interventions. The research has provided important practical benefits in terms of understanding the course of acute myocardial infarction and the potential effects of conventional therapies. For the present, interventions designed to limit infarct size remain in the realm of clinical research; routine clinical use would be inappropriate.


Pulmonary tumorigenesis in Syrian golden hamsters after intratracheal instillations with automobile exhaust condensate. Syrian golden hamsters were intratracheally instilled once every two weeks for life with automobile exhaust condensate (AEC) at two dose levels. The condensate was prepared from the most common German passenger car driven in the Europa-Test cycle (simulated city driving) and contained 340 microgram/g benzo(a)pyrene (B(a)P). Despite the very low total dose of B(a)P received, all animals of both dosage groups developed multiple pulmonary adenomas. Electron microscopy revealed these tumors to resemble B(a)P induced pulmonary adenomas in their ultrastructure. Numerous condensate-laden macrophages in tumor and peripheral lung tissues indicated that little of the instilled condensate had been removed from the airways through mucociliary action.


Expression of histocompatibility (HLA) antigens on tumor cells and normal cells from patients with melanoma. The expression of HLA antigens and beta2-microglobulin (beta2-mu) on cultured melanoma cells originated from 11 patients has been quantitated and compared with that on fibroblasts and cultured human lymphoid cells originated from the same patients. No qualitative or quantitative difference was detected with the exception of one melanoma line. HLA antigens were also quantitated in sera from melanoma patients: two sera reacted with anti-HLA-B7 antibodies although this specificity was not expressed on lymphocytes from whom the sera were obtained. A technique to quantitate HLA antigens on cells developed in the course of this study is described.


Eosinophilic meningitis complicating Hodgkin's disease. A report of a case and review of the literature. A case of eosinophilic meningitis as a complication of Hodgkin's disease is reported. Intrathecal chemotherapy with methotrexate and whole brain radiation therapy were effective in clearing the cerebrospinal abnormalities, pathologic evidence of meningeal disease, and the clinical manifestations. The differential diagnosis of eosinophilic meningitis and therapy of lymphomatous meningitis are discussed.


Detection of circulating tumor antigens. A wide variety of malignancies have associated tumor antigens. Two which have proven useful in the clinical detection and management of cancer are alphafetoprotein (AFP) and the carcinoembryonic antigen (CEA). These materials have been determined both by radioimmunoassay and by a newer technique employing enzymes to replace radioisotopea as markers. The interpretation of these immune assay results and the factors governing the shape of the standard curve are discussed. The advantages of 57Co over 22Na as a volume marker in radioimmunoassay are presented.


Recent advances in cancer diagnosis with nuclear medicine techniques. Recent progress in cancer detection in nuclear medicine has been significant and encouraging. Three main trends are responsible. First, techniques of imaging have improved, often in conjunction with the development of new and better imaging devices. Secondly, new radiopharmaceuticals are being investigated. Finally, interaction with other imaging modalities has started. In general, this yields complementary information further refining the diagnostic process.


Suppression of tumor-specific cell-mediated cytotoxicity by immunoregulatory alpha-globulin and by immunoregulatory alpha-globulin-like peptides from cancer patients. The suppression of tumor-specific cell-mediated cytotoxicity by human immunoregulatory alpha-globulin (IRA), by a peptide fraction derived from IRA, and by IRA-like peptides from the serum of cancer patients was studied in a syngeneic murine tumor-host system. Splenic lymphocytes from tumor-immunized mice were cytotoxic specific tumor cells in vitro as measured by the [125]-iododeoxyuridine release microcytotoxicity assay. However, this effect was significantly depressed if 1.25 to 5 mg of IRA per ml were added to the cultures. Pooled lyophilized normal human serum protein was inactive. IRA peptide and IRA-like peptide fractions from cancer patients were also highly suppressive of cell-mediated cytotoxicity at much lower concentrations (0.05 to 0.5 mg/ml). Control human serum peptide, which failed to inhibit the induction of hemolytic plaque-forming cells in sheep erythrocyte-injected mice, had no effect on cell-mediated cytotoxicity. IRA and IRA-like peptide fractions were not cytotoxic to the effector lymphocytes or to the target cells at the concentrations used.


Evidence for common and distinct determinants of colon carcinoembryonic antigen, colon carcinoma antigen-III, and molecules with carcinoembryonic antigen activity isolated from breast and ovarian cancer. This study was designed to answer the question, do molecules with carcinoembryonic antigen (CEA) activity from colon, breast, and ovarian cancer differ? Extracts of two breast and three ovarian cancers with CEA activity were compared to three colon cancer CEA preparations and to the related antigen, colon carcinoma antigen-III, in terms of lectin- and antiserum-binding properties. With the use of Farr-type radioimmunoassays with the lectins, concanavalin A and wheat germ agglutinin, the iodinated colon CEA and CEA-like preparations from breast and ovarian cancer all showed distinctly different patterns of binding. Specificity of binding was confirmed by inhibition studies with the relevant monosaccharides. Similarly, with antisera prepared against colon CEA, colon carcinoma antigen-III, or breast CEA, it was shown that, although all preparations shared some antigens, unique antigenic determinants were also present on all preparations. These data are consistent with the concept of a series of closely related CEA and CEA-like molecules with distinct characteristics for each tissue source of CEA.


Antigenic differences among osteogenic sarcoma tumor cells taken from different locations in human tumors. Sections were taken from the center, midzone, and margin of four human osteogenic sarcomas and one fibrosarcoma. Single-cell suspensions of tumors were examined in an indirect immunofluorescence assay with autologous or homologous anti-osteogenic sarcoma antisera as the intermediate reactant and fluorescein-labeled anti-human IgG as the final reactant. Cells were stained under conditions in which the fluorescence intensity was directly proportional to the density of the tumor-associated antigen on these cells. The density of tumor-associated antigen on cells from the center of the five tumor masses was low; cells from the midzone had intermediate levels of tumor antigen density, and cells at the margin had the highest levels. Similar preparations stained with polyspecific anti-HLA antisera did not demonstrate such a gradient. Since osteogenic sarcomas grow outward from the center, with the outer margin populated by the youngest cells, these results suggest that the oldest cells in the tumor bear the least tumor antigen, and the youngest tumor cells have the most. This is not compatible with theories which postulate that the immune system modulates the growth of a tumor so that only the least antigenic cells are allowed to grow. Alternative mechanisms are discussed.


Presence of DNA polymerase in lymphosarcoma in northern pike (Esox lucius). Northern poke lymphosarcoma DNA polymerase was partially purified from particulate fractions banding at 1.15 to 1.16 g/ml from homogenates prepared from frozen necropsies of tumor-bearing pike. The enzyme behaves as a typical reverse transcriptase, in that it prefers ribotemplates to deoxytemplates. The isoelectric point (pl 5.5) is similar to that of avian myeloblastosis virus polymerase. The pike enzyme elutes from a phosphocellulose column at 0.22 M potassium phosphate, the same as avian myeloblastosis virus DNA polymerase. The enzyme activity is inhibited by pyran, a specific inhibitor of viral DNA polymerases. The most striking difference between the pike lymphoma polymerase and the other viral DNA polymerases tested is the low maximum temperature of 20 degrees, compared to 30 degrees for Rauscher leukemia virus polymerase and 38 degrees for avian myeloblastosis virus and Rous sarcoma virus.


Biological and biochemical properties of Nonidet P40-solubilized and partially purified tumor-specific antigens of the transplantation type from plasma membranes of a methylcholanthrene-induced sarcoma. Tumor-specific transplantation antigen (TSTA) was solubilized from cell membranes of sarcoma Meth-A with non-ionic detergent Nonidet P40. Soluble TSTA was partially characterized by chromatographic separation and electrophoresis. The antigen responsible for tumor rejection activity had a molecular weight of approximately 70,000 daltons in the presence of detergent and an electrophoretic mobility of alpha-globulin. TSTA was well separated from mouse histocompatibility antigen H-2 by a sequence of procedures, including gel filtration, lectin affinity chromatography, column electrophoresis, and rechromatography on agarose, showed only three major bands on polyacrylamide gel electrophoresis. TSTA was specific for sarcoma Meth-A.


Dextran derivatives in single and combination chemotherapy against transplantable mouse ascites and solid tumors. Dextran, a typical homopolysaccharide without antitumor activity, was modified by palmitoylation and/or phosphorylation to yield three derivatives: palmitoyldextran phosphate, dextran phosphate, and palmitoyldextran. Of these compounds, only palmitoyldextran phosphate showed growth-inhibitory activity against Ehrlich solid tumor in mice. In combination therapy with mitomycin C, bleomycin, cyclophosphamide, and 5-fluorouracil, palmitoyldextran phosphate manifested strong synergistic effects against both Sarcoma 180 ascites and L1210 leukemic tumors. The compound is not directly cytocidal against Sarcoma 180 ascites tumor, but it appears to act via activation of peritoneal macrophage. The antitumor activity of palmitoyldextran phosphate apparently is mainly due to immunological host-mediated mechanisms.


[Effectiveness of intranasal immunization with Myxovirus influenzae neuraminidase in mice]. An intranasal immunization with a A/PR8/34-isolated NA, protected mice as well as the whole virus and A/Hong Kong/1/68 virus against a subsequent infection with mice-adaptated A/PR8/34 strain.


[Antigenic activity of lobster (Homarus vulgaris) arginine kinase and its cyanogen bromide fragments]. The antigenic saturation of lobster arginine kinase (38 000 daltons) by its specific antibodies has been studied. It was found that seven antigenic binding sites are simultaneously reactive on the surface of the enzyme in the presence of a large excess of antibodies or of their Fab fragments. After cyanogen bromide cleavage, the antigenic reactivity is distributed on several fragments of various sizes.


Re-entrant ventricular arrhythmias in the late myocardial infarction period. 4. Mechanism of action of lidocaine. The effect of lidocaine on re-entrant ventricular arrhythmias (RVA) was studied in dogs 3-7 days following ligation of the anterior descending coronary artery; direct recordings were made of the re-entrant pathway (RP) from the epicardial surface of the infarction zone (IZ). Lidocaine in a therapeutic dose consistently prolonged refractoriness of potentially RP(s) in the IZ and produced a higher degree of conduction block at a constant heart rate. Conduction in the adjacent normal zone was not affected. The impairment of conduction induced by lidocaine in the RP was directly related to its ability to abolish re-entrant ventricular beats and tachycardia. Gradual slowing of conduction in the RP consistently developed before abolition: lengthening of coupling of extrasystolic beats in surface leads and gradual slowing of ventricular tachycardia rate occurred. The termination of re-entry was characteristically associated with complete block in the RP. A "selectivity hypothesis" for the antiarrhythmic action of lidocaine is proposed.


Sinoatrial pacemaker shift following atrial stimulation in man. Indirect evidence of a sinoatrial pacemaker shift after programmed atrial stimulation in man is presented. Following electrically induced beats, time intervals and postextrasystolic morphology of atrial electrogram and P waves were scrutinized in 30 catheterization studies. Applying premature atrial stimulation, a decrease of the interval between the last basic atrial depolarization and the stimulus-produced atrial excitation (curtailed cycle) below a critical interval was followed by a sinoatrial pacemaker shift in three cases. This electrophysiologic event consisted of a concomitant change in shape of high right atrial electrogram and an increase of atrial cycle length. Simultaneous alteration of P waves could be detected in 2/3 patients. Assuming that the pacemaker shift indicates the arrival of ectopic activation in the sinus node, capture of the sinus node by the premature beat could be distinguished from failure to capture. Thus, pacemaker shift can be used for estimating sinoatrial conduction time in addition to present methods using measurement of postextrasystolic atrial intervals. The changes described could be seen both before and after atropine administration. Tracings of a pacemaker shift after cessation of rapid atrial pacing are also presented. In summary, we found a sinoatrial pacemaker shift underlying sinus node response to ectopic atrial activation in man, a phenomenon which contributes to our understanding of indirect assessment of sinoatrial conduction time by the premature stimulation technique.


The effect of left atrial-to-aortic assistance on infarct size. Left atrial-to-aortic (La-A) assistance is effective in supporting the failing circulation. This study evaluated its effect in salvaging ischemic myocardium in both large and small infarct models. In a control group, good correlation was shown between measurements of infarct size by ST-segment mapping at 20 minutes, triphenyl tetrazolium chloride staining at 5 hours, and the distribution of radioactive microspheres (P less than 0.01). A servo controlled assist pump was used which controlled pump speed according to the left atrial pressure. This allowed greater degrees of bypass for prolonged periods with reduced risk of air embolism. La-A assistance reduced systolic left ventricular pressure, and reduced the pressure time index (P less than 0.05). La-A assistance did not reduce infarct size measured by ST-segment mapping in a large infarct model when instituted before occlusion or 20 minutes after occlusion. Triphenyl tetrazolium chloride staining, however, showed less severe and homogenous damage in the assisted group as compared to controls. Electron microscopy confirmed the patchy nature of the ischemia. In a small infarct model, La-A assistance reduced nST from 6.3 +/- 0.8 to 3.8 +/- 1.5 and ST from 4.9 +/- 0.6 to 2.7 +/- 0.9 (P less than 0.05). The endocardial to epicardial ratio in the ischemic area fell from 0.69 +/- 0.05 to 0.43 +/- 0.05 in the control group (P less than 0.05) and a similar fall occurred in the assisted groups. La-A assistance is thus effective in reducing myocardial ischemia in a small infarct model, but appears to be less effective in a large infarct model.


Serum UDP-galactose: glycoprotein galactosyltransferase in diabetics with microangiopathy. 1. Levels of serum UDP-galactose:glycoprotein galactosyltransferase in 117 unselected diabetics were compared with those in 60 non-diabetic healthy controls. 2. Enzyme activity (mean +/- 2 S.D.) of control sera was found to be 90.2 +/- 21.5 etamoles/ml/hr at 37 degrees. In 30 of the 117 diabetic sera (26%) enzyme activity was elevated (greater than mean + 2 S.D. of the controls). Sixteen of 19 (84%) patients with retinopathy, 16 of 26 (62%) patients with peripheral vasculopathy and 13 of 26 (50%) patients with neuropathy had higher levels of serum enzyme. When serum enzyme levels of groups of diabetics with retinopathy, peripheral vasculopathy and neuropathy were compared with the enzyme level in all diabetics, there was a significant difference with p values of 0.001, 0.05 and 0.05 respectively.


Serum beta2-microglobulin in controls and cancer patients. Serum beta2-microglobulin levels have been measured in 210 cancer and control patients to assess the significance of this investigation in cancer patients. Subjects studied included patients with breast and gastrointestinal cancer, corresponding control patients in both categories, and healthy volunteers. The composition of these groups allowed an assessment of the relative importance of changes related to cancer, benign disease, age and sex. A significant rise in serum beta2-microglobulin levels with advancing age was demonstrated in the control subjects. Mean levels were also consistently higher in females than in males in each patient group. After statistical correction for these age and sex effects, mean values remained significantly higher in each of the various cancer groups than in their controls. Patients with more advanced breast cancer had higher levels than those with 'early' disease, as did patients with stomach cancer compared to those with colo-rectal cancer. One possible interpretation is that levels increase with increasing tumour bulk, and therefore the estimation of serum beta2-microglobulin may be useful as one of a battery of tests in the management of cancer patients.


[Production of interferon and alpha 2-macroglobulin involvement in immune response during human viral hepatitis A (author's transl)]. In viral hepatitis A, we could distinguish between monophasic and polyphasic forms. Monophasic and polyphasic hepatitis A induce the production of plasma interferon. Interferon level, elevated as early as the first days following the appearance of clinical signs, decreases and reaches a minimum value on the seventh day. A new rise of interferon is characterized by a maximum level on the twelfth day and a minimum level on the thirtieth. Beyond the first month we could still detect the presence of interferon. In the two forms of hepatitis, a complement system is activated both by classical and alternate pathways. IgM levels increase early, IgA levels remain unchanged. On the other hand, IgG levels, only slightly elevated in monophasic hepatitis A, are highly increased in polyphasic hepatitis A beyond the first month. Alpha 2-macroglobulin reaches levels above normal during convalescence in monophasic hepatitis A; on the contrary, in polyphasic hepatitis A, alpha 2-macroglobulin levles are above normal as early as the thirty first days of illness and remain above normal for several months. Elevated levels in alpha 2-macroglobulin may inhibit cellular immunity which is accountable for immunological injury of virus infected hepatocytes. We wonder whether this earlier increase in alpha 2-macroglobulin is responsible for the lasting character of viral infection observed in polyphasic hepatitis A.


The determination of serum immunoglobulin concentrations on the basis of their light-chain antigenic properties. A method for the measurement of light-chain determined immunoglobulin concentrations (Ig-type  kappa and Ig-type lambda) in serum is described. The method is based on the principle of radial immunodiffusion. The results obtained after calibration with standarized human serum and with isolated Bence-Jones protein are closely correlated. The values for Ig-type kappa, Ig-type lambda and the kappa/lambda ratio from sera with normal concentrations of IgG, IgA and IgM are presented and some problems of calibration discussed.


Abnormal breakdown of alpha2-macroglobulin-trypsin complex in cystic fibrosis. The complex of trypsin with purified alpha2-macroglobulin from normals and patients with cystic fibrosis was studied. The formed complex failed to reveal any proteolytic activity toward a high molecular weight substrate whereas the esterolytic activity towards a low molecular weight substrate was retained. This esterolytic activity was resistant to inhibition by a high molecular weight inhibitor. During iincubation at 38 degrees C the complex with normal alpha2-macroglobulin was slowly inhibited by the high molecular weight inhibitor and regained activity with the high molecular weight substrate. This phenomenon was not obtained when the alpha2-macroglobulin from cystic fibrosis was examined. These data suggest that the gradual conversion of normal alpha2-macroglobulin-trypsin complex into an alpha2-macroglobulin fragment-trypsin complex is deficient in patients with cystic fibrosis.


Kinetics of the inactivation of human and bovine trypsins and chymotrypsins by alpha1-proteinase inhibitor and of their reactivation by alpha2-macroglobulin. The time dependency of inactivation of human cationic trypsin and chymotrypsin II and of bovine trypsin and alpha-chymotrypsin by human serum has been investigated. Since the molar concentration of serum alpha1-proteinase inhibitor is much higher than that of other inhibitors, this time dependence could be used to calculate the rate constants kass for the association of alpha1-proteinase inhibitor with the four proteases. The association process was found to be second order, with kass ranging from 1 x10(4) s-1 (human trypsin) to 2.6 x 10(6) s-1 (bovine chymotrypsin). The human proteases react much more slowly with human alpha1-proteinase inhibitor than the bovine ones. But, whatever the species, chymotrypsin is inhibited more quickly than trypsin. Addition of alpha2-macroblobulin to the inactive complexes resulted in a time-dependent regeneration of enzymic activity due to the formation of alpha2-macroglobulin-protease complexes. The reactivation (i.e. dissociation) process was first order and extremely slow: the half-life of the alpha1-proteinase inhibitor-proteinase complexes ranged from 8 days (bovine chymotrypsin) to 9 months (human chymotrypsin). The human proteases formed the most stable complexes with alpha1-proteinase inhibitor. The pathological implications of these findings are discussed.


Ag-staining of nucleolus organizer regions of chromosomes after A-,C-, G-, or R-banding procedures. Metaphase chromosome preparations were made from leukocyte cultures of normal individuals. The cells were fixed in methanol:acetic acid (3:1 v/v), then dropped on cold, wet slides which were air-dried before storage at 4 degrees C. The slides were stained to identify the chromosomes by one of the following procedures: (1) Quinacrine. Slides were stained for 10 min in quinacrine mustard solution, rinsed in running tap water for 2 min, and mounted in Tris-maleat buffer, pH 5.6.


Q- and C-bands in the metaphase chromosomes of Drosophila nasutoides. The large heterochromatic chromosome of Drosophila nasutoides reveals distinctive C- and Q-bands. The regions which are negative in C-banding appear positive in Q-banding. The isochromosomic nature of this chromosome and the locality of the satellite DNAs in this chromosome are discussed with respect to these banding patterns.


The effectiveness of post-detoxication referrals: effects on later detoxication admissions, drunkenness and criminality. This study concerned the effectiveness of post-detoxication referrals to a variety of treatment facilities. The purpose was to discover (i) the proportion of men accepting referrals who actually arrived, (ii) the differences in outcome for patients attending an out-patient clinic, a halfway house, and a long-stay farm, (iii) the differences in outcomes for patients treated in one of the above facilities compared with those for similar patients not receiving treatment. In all, 114 male detoxication admissions were included. However, only 60% arrived, even when firm referrals were made. Those arriving and not arriving did not differ in social or demographic characteristics, nor did those who were referred to the various treatment facilities. However, the referral group had more detoxication admissions in the post-detoxication period. There were no post-treatment overall differences between all treated and untreated patients in detoxication admissions, arrests for drunkenness or criminal convictions. Refusals were more often successes than the treated or untreated groups but this was due to their better prognosis at intake. In general, the data provide little cause for optimism about the value of post-detoxication referrals.


[Thiamine, riboflavine and pyridoxine supply in chronic alcoholism (author's transl)]. Tests on 50 chronic alcoholics demonstrated that there was a reduced supply of thiamine (measured with the transketolase activation test) and riboflavine (glutathione reductase test) compared with results obtained in 1152 healthy controls. A high risk of thiamine deficiency was present in 45.4% of male and 50% of female alcoholics; risk of riboflavine deficiency was 11.1% and 16.7%, respectively. Corresponding results in the normal controls were 2% for thiamine and 0.4% for riboflavine deficiency. There was a significant relationship between enzymatic values of thiamine deficiency, on the one hand, and anaemia, abnormal liver functions (bilirubin, gamma-globulin) and low diastolic arterial pressure, on the other. Enzymatic riboflavine values also correlated with haemoglobin content, so that it is assumed that the anaemia is associated with the reduced thiamine or riboflavine supply. Enzymatic determination of pyridoxine with the GOT activation test would seem to be influenced by alcohol or by metabolic processes induced by it and is therefore not suitable in the presence of chronic alcoholism.


Early evoked potentials in patients with acoustic neuroma. Using clicks with varying interstimulus intervals and coherent tone-bursts, early components of the auditory evoked potential (brain stem responses) were studied in four patients with confirmed acoustic neuroma. Abnormalities in responses appeared with shorter interstimulus intervals and with tone-bursts delivered monaurally to the involved ear; bilateral alterations occurred in one patient with brain stem displacement. The results indicate that BSR can provide a stable, independent, noninvasive measure of auditory nerve function useful in the early detection of acoustic neuroma. The results contribute to the understanding of the so-called human FFR.


[Somatosensory evoked potentials and Hoffmann reflex in acute spinal cord lesions; physiopathological and prognostic aspects]. Twenty-four patients with recent and acute spinal cord lesions were examined. The somatosensory cerebral evoked potential (SEP) following stimulation of the peroneus communis nerve tested spinal conduction, whereas the H reflex showed spinal excitability belowe the lesion. After complete spinal cord section, the SEP was always abolished and the H reflex was absent in most cases tested in the first 24 hours. Into other patients, the recruitment curve and the recovery cycle of the H reflex displayed some abnormalities which progressively disappeared. With partial lesions, SEP could occasionally be altered. Some abnormalities of the H reflex recovery cycle, of the same type as those seen in the late stage of complete sections, were also observed. These data give nre information on the physiopathology of spinal shock; they lead to the distinction of several evolutionary stages after acute spinal lesions and also have prognostic disgnificance.


Cerebral cortical isolation in infantile neuroaxonal dystrophy. Three patients with infantile neuroaxonal dystrophy (INAD, Seitelberger's disease) studied between ages 2 and 5 years, with the characteristic electroencephalographic pattern of high-voltage, fast (16-24 c/sec) rhythms and absence of reactivity on eye-opening or closure (Radermecker and Dumon-Radermecker 1972), also showed no changes in response to intermittent photic stimulation and absence of evoked potentials to flashes, clicks or median nerve stimuli. Although some theta rhythms and delta activity appeared during drowsiness and sleep, the fast rhythms persisted as the dominant feature. There were no central transients or K-complexes. When the patients cried, with hyperventilation, and also during breath-holding spells, slow rhythms appeared and the fast rhythm was reduced. These findings are interpreted as evidence of cerebral cortex isolation, the fast rhythm representing the spontaneous ("idling") activity of the cortex largely disconnected from subcortical or remote cortical influences by the slowly progressive, selective degeneration of axons, characteristic of the pathology of INAD, but the cortex remains responsive to chemical influences. These electrophysiological features become established furing the third year of age, prior EEGs being normal and later ones showing paroxysmal and other abnormal features in addition to the fast rhythm.


Sleep patterns in mental retardation: Down's syndrome. EEG and eye movements during two consecutive nights were recorded in 21 "normal" and 21 Down's subjects of both sexes in the 14-24-year age range. Records of most Down's subjects were characterized by longer total sleep time, lack of waking alpha, frequent awakening and movement epochs and fewer spindle bursts. Significantly longer "awake" and stage 4 percentages, but less stage 2, were found in the Down's group. They also had longer REM latency, and lower eye movement density based on number of eye movements. Density based on integrated eye activity correlated positively with IQ in the Down's subjects. Spectrum analysis for EEG frequencies from 1 to 20 c/sec showed significantly greater power in the Down's group below 9 and above 12 c/sec, but no difference in the alpha range. The greater power at low frequencies was more pronounced in "awake", stage 1 and REM.


Visually evoked potentials from occipital and precentral cortex in visually deprived humans. The visually evoked potential (VEP) was recorded from the scalp overlying precentral and occipital cortex in three monocularly deprived humans. The subjects had defects of the globe from birth that effectively created a condition of diffuse light rearing in one eye. In subject, BER, a 19-year-old with a recently removed congenital cataract, the occipital response evoked from the deprived eye was reduced by 53% compared with the response evoked from the good eye. In subject GUZ, age 20 with a congenital cataract, the occipital response evoked from the deprived eye was actually larger by 20% than was the response evoked from the good eye. For UTZ, age 5 with whitish cellular debri in the anterior vitreous of the eye, the occipital response evoked from the deprived eye was only 4% smaller than the response evoked from his good eye. In all three subjects, the precentral response evoked from the deprived eye was reduced in amplitude compared with the precentral response evoked from the good eye. In terms of relative effect of the deprivation upon the VEP from the two recording sites, the precentral VEP was altered to a greater extent than was the occipital VEP. These findings indicate that diffuse light rearing can affect the pathways projecting to precentral cortex independently of the specific visual pathways.


Ontogeny of delta activity during human sleep. The study describes the ontogenetic and time-related changes in peak amplitude, frequency, and incidence of delta waves of twenty-five normal subjects between the ages of 3 and 79 years of age. All slow waves greater then 5 muV in magnitude and between 0.5 and 3.0 c/sec were analyzed for the first 6 h of the night's sleep. In this study increasing age was accompanied by decreases in average peak amplitude of delta waves over 5 muV, slowing of delta frequencies, and decreases in incidence of waves greater than 20 muV. Across 2-h epochs of the night there were tendencies for the average peak amplitude of delta waves to decrease, for the average frequency for a given amplitude range to decrease, and for the incidence of low-amplitude delta waves to increase and high-amplitude waves to decrease. The incidence of waves greater than 5 muV remained constant across ages and epochs. Frontal channel delta waves exhibited an age-group difference whereas no such difference was apparent for central channel delta activity, suggesting that the frontal channel may possess advantages over the central because of greater sensitivity to age related changes.


Effects of cooling on guinea pig olfactory cortex maintained in vitro. Electric activities from brain slices of guinea pig olfactory cortex were studied during gradual cooling without a temperature gradient. At normal temperature the potential evoked by stimulation of the lateral olfactory tract consists of an initial spike (IS), a negative (N), and a positive (P) potentials. The IS potential has been considered to be presynaptic in origin and the latter two postsynaptic. During cooling from 37 to 15 degrees C, the amplitude of both the IS and the N potentials increased to attain the maximum value (10-36% increase) at about 32 degrees C, then decreased gradually and disappeared at around 15 degrees C. On the other hand, the durations of the IS and the N potentials were prolonged gradually on cooling from 37 to 17 degrees C. Most of the olfactory cortical neurons responded with one or two firings to the tract stimulation. The firing numbers increased on cooling down to about 32 degrees C and declined on further cooling, which corresponds with the behavior of the N potential. All these effects were reversible on rewarming. Augmentation of the N potential might depend primarily on the increase in amplitude of the IS potential and secondarily on the increase of amount of liberated transmitter substance and/or the delayed inactivation process of the transmitter action.


Duration of paradoxical sleep episodes. A quantitative and pattern analysis of reticular multi-unit activity in the cat. This study was aimed at the possible relationships between the level of brain activation in PS and the duration of episodes. Multi-unit activities from 16 recording sites in the mesencephalic and pontine structures were quantitatively evaluated in five cats and the influence of electrode localization and episode duration on the average patterns of integrated discharges was statistically investigated. The analyses emphasized the slow time course of the early increments in reticular activity and its culmination at a plateau level in the later part of PS phases. Differences in mean amplitude of this plateau were related to the anatomical localization of electrodes but no distinction could be made between reticular areas with respect to the temporal features of activation patterns. Synchronization of phasic bursts was consistently observed between simultaneously recorded structures and this, along with the time course of early increments in SW was consistent with the notion that mesencephalic reticular activity is controlled by leading influences from the pons. No basic differences in the temporal development of reticular activation in PS were found in relation to the duration of episodes.


Sleep patterns in the "locked-in" syndrome. Sleep patterns in two chronically "locked-in" patients were studied. The first patient had sleep activity similar to other quadriparetic patients with reduced physical activity. The second patient experienced no REM sleep and only short bursts of non-REM sleep. Clinical evidence suggested extension of his lesion dorsally into the pontine tegmentum. This correlates well with experimental evidence indicating that structures in the tegmental area are essential for normal sleep.


[Schizophrenia: association of the paranoid form and antigens HLA-A9 and B5]. 30 unrelated Schizophrenic patients (15 Hebephrenic and 15 Paranoïd forms) were Typed for 14 A and 16 B Specificities by microlymphocytotoxicity. We note: 10 (5H + 5P) HLA--A1 (33.3 % / controls 23.4 %) 5 (2H + 3P) HLA--A3 (16.6 % / controls 26.2 %) 6 (5H + 1P) HLA--B8, 0 HLA--B27, 0 HLA BW16 and more specially: 10 (3H + 7P) HLA--A9 (23 + 24) 8 (2H + 6P) HLA--B5 These two last antigens are more often found in the Paranoïd patients of whom 46.6% are A9 Tcontrols: 24 %) and 40 % are B5 (controls 13.2 %. The Pc values are not significant in these too short seria but the strongly increased frequencies of A9 and B5 in the Paranoïd group seem to open a new biological aspect in the nosographic distinction of the Schizophrenia and so confirm previous results of C. L. CAZZULLO et al.


Serological studies with the ficin-antificin system. Rabbit antisera prepared against ficin reacted with it in a series of serologic tests. Upon immunodiffusion analysis, ficin was found to consist of at least nine antigenic components. Ficin will adsorb spontaneously to erythrocytes which can be used in passive hemagglutination tests. The enzymatic activity of ficin was not abolished by antificin sera.


Evidence for selective axon-terminal uptake and retrograde transport of label in cortico- and rubrospinal systems after injection of 3H-proline. In tracing the macaque's ascending spinal connections by anterograde axonal flow, perikaryal labelling was also observed within the n. ruber pars magnocellularis as well as in the sensorimotor cortex. While neuropil labelling was demonstrated after single injections of leucine or proline, labelling of perikarya was only evident following proline injections. Perikaryal labelling is most likely to be due to uptake of label by axons and/or axons terminals within the injection field and subsequent retrograde axonal transport of radioactive compound to the parent cell bodies. Advantages and disadvantages of the uptake of label by axon terminals are discussed in relation to the use of 3H-proline for tracing neuronal connections. The selectively of this event and its relationship to the suggested inhibitory functional role of proline and its metabolites is not clear and needs further investigation.


The effects of monocular deprivation on different neuronal classes in the lateral geniculate nucleus of the cat. Retrograde axonal transport of horseradish peroxidase (HRP) was used to identify two populations of cells in the lateral geniculate nucleus (LGN) of the cat. HRP was injected into area 17 and 18 separately in the same animals, and the neuronal somata giving rise to thalamo-cortical axons, identified by the presence of granular HRP reaction product within them, were measured. The mean size of LGN neurones labelled by injections in area 17 ("17-relay" cells) was less than of neurones filled from area 18 ("18-rely" cells). Similar separate injections into area 17 and 18 of monocularly deprived kittens also showed that in non-deprived LGN laminae 17-relay cells were, on average, smaller than 18-relay cells. In deprived laminae, 17-relay cells were some 20% smaller than in non-deprived laminae, but deprived 18-relay cells were 50-60% smaller than normal, being on average, actually smaller than deprived 17-relay cells. We conclude that the population of large LGN neurones projecting to area 18 is more severely affected by monocular deprivation than the smaller neurones projecting to area 17, and discuss the relationship of the morphological results to physiologically defined X and Y cells in the LGN.


[Relationship between the endocochlear potential and metabolism in the vascular stria of the inner ear]. Effects of dihydrostreptomycin, etacrine acid and ATPh on the endocochlear potential in guinea-pigs depended on the dosage and means of administration. Dihydrostreptomycin suppressed the potential only when perfusing the perilymphatic space or being microinjected into the endolymphatic duct. I. v. administration of etacrine acid suppressed the potential with its following partial restoration. I.v. administration of ATPh in large doses suppressed the potential for a short period but did not prevent the ototoxic effect of etacrine acid. ATPh in small doses did not suppress the potential but exerted the protective effect.


A syndrome of functional hypogonadotropic hypogonadism and sterility in a male with elevated serum estradiol. A 36-year-old male complaining of impotence was examined. He was a genotypic male. Phenotypically, he exhibited signs of long-standing estrogen excess, such as feminine body build, gynecomastia, and varicose veins. His testes were soft and borderline small, and his prostate was small and soft. However, he had a normal-sized penis, normal male hair distribution, normal sense of smell, and normal intelligence. The laboratory data were compatible with mild hypogonadotropic hypogonadism. Serum estradiol (E2) levels were consistently elevated. The patient had azoospermia and a decreased semen volume. Inappropriately low levels of luteinizing hormone and follicle-stimulating hormone responded normally to gonadotropin-releasing hormone and clomiphene citrate. Levels of both testosterone (T) and E2 increased dramatically after prolonged clomiphene medication and in response to human chorionic gonadotropin. There was no change in either T or E2 levels in response to manipulations of the pituitary-adrenal axis. It is concluded that the elevated E2 level was responsible for suppression of gonadotropins which, in turn, caused mild hypogonadism and sterility in this patient. According to the stimulation tests, the source of the elevated E2 levels was testicular.


[Abdominal pregnancy: hormon concentrations during the postpartal period with placenta remaining intra-abdominal (author's transl)]. It is reported of an abdominal pregnancy at term. The placenta was left in situ because of the high risk of intraabdominal bleedings. Thereby it was possible to measure hormon concentrations of the fetoplacental unit without the fetal compartment. Within 10 days after delivery we determined the plasma levels of estradiol-17 beta, estriol, progesterone, HCS, alpha1fetoprotein, and the excretion of the total estrogens in the urines. There was near the same decrease of hormon concentrations in the post partal time comparable with that of a normal pregnancy. Only the HCS concentrations didn't change in the first 9 days after delivery.


Nucleolus organizers in Mus musculus subspecies and in the RAG mouse cell line. Silver staining has been used to detect active nucleolus organizer regions (NOR's). By this criterion six mouse chromosomes, numbers 12, 15, 16, 17, 18 and 19, can have an NOR. The number and distribution of chromosomes with NOR's vary among inbred strains of Mus musculus musculus (C57BL/6J, BALB/cJ, C3H/HeJ and C3H/StCpr1BR) and in M. musculus molossinus. In a musculus X molassinus F1 hybrid, nucleolus organizers from each parent are silver stained.--Chromosomes which have NOR's in diploid cells also show them in tetraploid cells and in established cell lines. The BALB/cJ strain shows Ag-staining of NOR's on chromosomes 12, 15, 18 and occasionally 16. In the RAG cell line, which was derived from BALB/c, active NOR's are seen on 12, 15 and 18, even after these chromosomes have undergone structural rearrangements in the cell line. Some correlation exists between the amount of Ag-stain and the size of a secondary construction region, with a large amount of Ag-stain present on a chromosome which has a prominent secondary constriction. There is no correlation between the amount of Ag-stain and the presence or absence of C-band material.


[Review: hemorrhagic diathesis resulting from acute exposure to ionizing radiation]. The symptoms of the acute radiopathy are chiefly characterized by a severe blood coagulation disorder. The main results and problems of research work on this haemorrhagic diathesis are shortly reviewed.


[A case of Osler-Rendu disease with simultaneous thrombopenia and a factor VIII inhibitor]. Long-term determinations of haemostasis factors in a case of Osler's telangiectasia revealed the temporarily simultaneous existance of periods of thrombocytopenia, a decrease of prothrombine and a reduction of the fibrinogen and plasminogen level. These findings considered as signs of consumption coagulopathy coincided with an increased bleeding tendency of the patient. The correlation existing between the clinical symptoms and the observed disorders of coagulation may possibly be explained by the appearance of an intravascular coagulation during the late period of the haemorrhagic diathesis, which could be proved by the simultaneous increase of fibrinogen degradation products. Moreover, the patient's plasma was capable of strongly inhibiting factor VIII of a normal plasma. The possible influence of plasmatic disorders of coagulation which are caused by secondary reasons on the clinical picture of a haemorrhagic diathesis primarily based on vascular conditions is discussed.


[Precision control of a single step determination of blood coagulation factor VIII in the plasma]. The findings of a precision control are reported for a one-stage determination of the factor VIII activity in the plasma for a period of 6 months. The storage property of the reagents was achieved by storing them in fluid nitrogen. Coefficients of variation under 55 were achieved at controlling the normal range as well as the pathological one. Thus the precision of this method may be compared with that of clinicochemical examinations.


[Leukocyte lysozymes in various forms of leukemia]. The amount of lysozyme in the leukocytes of 47 patients with different forms of leukaemia and 6 healthy persons was investigated. The lysozyme determination was carried out in the lysate of isolated leukocytes obtained after freezing and thawing it seven times. The results expressed in microgram per 10(6) cells were compared with the simultaneously determined lysozyme concentration of serum and urine. A substantial reduction of the lysozyme amount as compared with the normal value (3.1 microgram/10(6) cells) was determined in the leukocytes of patients suffering from chronic lymphatic leukaemia, acute lymphatic leukaemia and the blastic crisis of chronic myeloid leukaemia. Different amounts of lysozyme ranging from extremely low ones to strongly elevated ones were found in leukocytes taken from patients with acute myeloblastic and chronic monocytic leukaemia. In many cases there was a lack of correlation between the lysozyme content of leukocytes on the one hand and that of serum and urine on the other hand. Possible causes underlying this lack of correlation are discussed.


Heat-ethanol isolated serum albumin: discussion. Albumin isolated by the Hagen/Münster heat-ethanol method is a remarkably stable solution of high purity and clinical compatibility. The simplicity of this method makes it a model fractionation procedure for the small transfusion service. Immunization studies carried out by us are compared with previous accounts of a "new" antigen occuring in plasma protein solution produced by partial heat denaturation. Clinical compatibility of cold-ethanol and heat-ethanol fractionated albumin solutions are also shortly discussed.


Anti-HBbrno: a new CIEP hepatitis B marker, more sensitive than HBsAg radioimmunoassay. Evidence is presented for the new HBbrnoAg-Anti-HGbrno system association with the HBsAg. The association is displayed particularly in a prevalence of Ausria detectable HBsAg carriers among the anti-HBbrno carriers, suggesting the anti-HBbrno to be a marker of contact with hepatitis B or, possibly, an indicator of co-existing HBsAg antigenaemia, not detectable by radioimmunoassay. Identity of HBbrnoAg with HBsAg/a, d, y, w, is ruled out, the identity with the other HBV associated antigenic specificities remaining questionable.


Juvenile pernicious anemia associated with intestinal nodular lymphoid hyperplasia and immune deficiency state. Report of a case. A 17-year-old girl with humoral immune deficiency state was observed, in whom pernicious anemia developed one year before her death. The patient died after a very severe diarrheal attack and respiratory tract infection. The necropsy revealed intestinal nodular lymphoid hyperplasia. The immunological evaluation showed depressed IgG and IgM concentration with lack of IgA, however in the patient's blood there were found B lymphocytes containing IgA immunoglobulin. On the effect of crude gastric antigen the patient's lymphocytes showed M.I.F. release, although there were no circulating autoantibodies.


[Correlation of clinical and anatomical findings in childhood meningitis]. Clinical and anatomical findings of 76 prophylactically not irradiated children with acute leukaemia were compared. In the group of 17 children (22%) with clinically manifest symptoms of CNS involvement in the course of the illness leukaemic infiltrates were identified in the CNS at the autopsy and during histopathological examinations even in 70% in spite of clinical healing and normalization of the cerebrospinal fluid finding. In children who had either no clinical symptoms of CNS leukaemia (42 children) or only terminal symptoms in the last 24 hours of their life (17 children) leukaemic infiltrations were found in 25.4% of the cases. Non-myeloid types showed 44.5% of histopathological changes, myeloid types 22.7%. Clinical main symptoms and anatomical findings were correlated to the corresponding groups of patients and typical illustrations were added.


[Clinical trial with long-term therapy of generalized plasmacytoma]. The authors report on findings in long-term therapy made by means of a combination of cyclophosphamide as attack dosis (partially also with polychemotherapy--COP, COPP-scheme) and double plasmapheresis. Since 1967 33 patients have been treated in this way. A group (6 patients) only received cyclophosphamide in a attack therapy of 15...25 mg/5g per body weight; a second group of 14 patients received the same dosis in combination with a double plasmapheresis. The third group of 13 patients in an advanced stage of the illness was treated polychemotherapeutically according to various schemes (COP-cyclophosphamide, vincristine, prednisone; COPP with Natulan) likewise in combination with double plasmapheresis. The observations made for 4 years in the two groups first mentioned showed favourable results in the second group with an average survival time of 35 months. In the third group only experiences of two years can be reported and thus a final answer cannot be given. However, it can already be stated that a clinical success requires the cytostatic therapy to be continued for a long time in combination with plasmapheresis.


[The suppressive effect of antilymphocyte globulin (ALG) on the formation of anti-ALG antibodies]. The quantitative follow-up of precipitin formation against IgG aids in investigating the question whether ALG when applied to the organism tends to suppress the immune reaction against ALG. Rabbits locally immunized with pig anti-rabbit ALG were repeatedly treated i.v. with the same ALG, and to control groups normal pig IgG and NaCl saline was administered. It was found that antilymphocytic antibodies greatly suppressed the precipitation formation against IgG molecules. In the later stages of application this effect became more pronounced, evidently due to the specific suppression induced by long-term administration of relatively high doses of antigen. A possible improvement in the prevention of precipitin formation in ALG treated patients, i.e. substitution of currently applied tolerogenic dose of normal IgG by a similar dose of ALG is suggested.


Haematological studies in relation to environmental temperature and different periods of breeding cycle in an air breathing fish, Heteropneustes fossilis. Investigations were made on different haematological parameters in H. fossilis at two environmental temperature (18 degrees C and 32.5 degrees C) and different periods of breeding cycles. The RBC counts 302.0-10(4)/mm3), WBC counts (10.6-10(4)/mm3), Hb concentration (19.0g%), haematocrit (36.0%), M.C.H.C. (57.7%), total blood volumes (0.966 ml) and per cent blood volumes (1.542%) gave significantly high values but M.C.V. (119.2 micrometer3) and erythrocyte surfaces (52.96 micrometer2) gave low values at higher temperature (32.5 +/- 1 degrees C) and during breeding period (G.S.I. = 10.4 +/- 0.52 g). The higher values obtained for different haematological parameters during breeding period at high temperature may be probably related to greater physical activity and higher metabolic activity at the time of reproduction.


Some aspects of haematology of an Indian grass snake Natrix stolata in relation to sex and size. Studies have been made on the erythrocyte counts per mm3 of blood, total differential leucocyte counts per mm3 of blood, concentration of haemoglobin per 100 ml of blood, haematocrit values and nucleocytoplasmic ratio in a common Indian grass snake, Natrix stolata, in relation to body weight of both the sexes in the same month of the year (during breeding phase). Logarithmic transformations have been used in case of erythrocyte counts and haemoglobin concentrations. From log/log calculation it seems that the correlation between body weight and erythrocyte is more prounounced in females than in males. The concentration of haemoglobin shows a very high degree of correlation in both the sexes. The haematocrit values show a clear trend of increase with increasing body weight in both the sexes. The nucleocytoplasmic ratio does not show much variation in different weight groups whereas in females the variation is well marked in different weight groups and the size of erythrocyte also becomes larger in higher weight groups. In both the sexes the leucocyte counts have been found that along with the increase in body weights, the number of leucocyte increases in juveniles but in adults the leucocyte number decreases with the increasing body weights.


[Agglutination of the red blood cells after experimental glycolytic alteration]. The present findings proved Alcian Blue as hemagglutinin suitable for the specific demonstration of negatively charged sites of the cell surface. Erythrocytes with reduced surface charge because of previous trypsination or desialization exhibited decreased Alcian Blue agglutination. On the other hand, Alcian Blue agglutinates more intensely red blood cells with a high surface charge (reticulocytes). Findings with A antiserum or the lectins prepared from Lens culinaris and Helix pomatia indicate a masking of glycolipidic group A receptors by glycoproteins of the erythrocyte membrane. Effects of enzymatic degradation (neuraminidase, trpysin, pronase) combined with incubation in NaF-medium for the deprivation of ATP could be explained by a rearrangement of receptors resulting in altered agglutinability of erythrocytes.


[Alcian blue induced mixed cell agglutination--short communication]. Because of its specific affinity to negatively charged sites of the cell surface, alcian blue is suitable to effect mixed cell agglutination between human erythrocytes and peritoneal macrophages of guinea pigs. This effect could be inhibited by adding heparin molecules which were negatively charged as well.


[A test system for the measurement of cellular RNA-synthesis in the human bone marrow]. A test for the cellular RNA-synthesis (incorporation of 3H-uridine in the RNA) of human bone marrow has been standardized with respect to the time of incorporation, the number of cells and the concentration of 3H-uridine. The following parameters were estimated for 500 microleter standard assay and 100 microleter aliquots for the determination of the radioactivity: time of incubation 80 min, number of nucleated cells 8 - 10(5), concentration of 3H-uridine 8,3 - 10(-6) M. Actinomycin D inhibits the RNA-synthesis to 90% in a concentration of 1.2 - 10(2) microgram/ml. The test appears generally applicable for the determination of the vitality of bone marrow after cryopreservation, the testing of cryoprotectants and haematotoxic substances and the control of the reaction of the bone marrow during chemical- or irradiation treatment of tumors.


Activity of beta-glucuronidase in peripheral blood lymphocytes of patients with cancer of the larynx. In 20 untreated male patients with cancer of the larynx, aged 35 to 55 years, the significant increase in the absolute count of beta-glucuronidase-positive lymphocytes in the peripheral blood was examined by means of the cytochemical method of Hayashi et al. (1964). The increase was due to an elevated absolute count of lymphocytes exhibiting the granular-diffuse and the diffuse enzymatic reaction; no significant changes were observed with regard to lymphocytes with the granular type of reaction. The authors discuss the significance of their observations for the evaluation of lymphocyte immune response against tumour specific antigens in patients with cancer of the larynx.


Intracellular enzymatic response of lymphocytes and neutrophils in patients with cancer of the larynx. In 20 men, aged 35 to 55 years, with untreated cancer of the larynx activity of lysosomal acid phosphatase (AP), beta-glucuronidase (GR) and N-acetyl-beta-glucosaminidase was determined cytochemically in peripheral blood lymphocytes and neutrophils by means of Barka and Anderson, Hayashi et al. and Hayashi's method, respectively; the results obtained were compared with those in 20 healthy men aged 20 to 30 years. Total count of GR-positive lymphocytes was higher in the patients than in normal persons. Total counts of AP-, GR-, and GS-positive lymphocytes with not disrupted enzyme-positive lysosomal granules within the cell cytoplasm were significantly lower and total counts of cells exhibiting the disruption of lysosomal granules and the diffuse type of cytochemical reaction were significantly higher in the patients when compared with the control group. The response of neutrophils consisted of a significant elevation in numbers of AP-, and GS-positive cells; overall score of enzyme activity studied in neutrophils was not altered in the patients. The authors disucss the significance of their observations in the light of data on participation of lymphocytic and neutrophilic lysosomal apparatus in the immunological response against tumour specific antigen in patients with cancer.


A peculiar lymph node anti-carcinoma reaction. A case of a laryngeal extensive carcinoma determining a severe obstruction is reported, where the development of a monstrous adenopathy is followed by the regression of the initial tumour until complete laryngoscopical disappearance, as well as of obstructive and general symptoms. Cytological and histological examinations of the above-mentioned lymph node demonstrated (besides metastatic cells) a peculiar cellular reaction, similar to rhinoscleroma, which represented an original anti-carcinoma defensive mode. The medical importance of this very rare case is discussed and the probable immune origin of this reactive cellular complex is explained by analogy with "Hodgkinom", where granuloma are the immune response of the body against Sternberg-Reed cell.


[The use of sterile nursing in hematology]. By means of own experiences gathered in the treatment of 18 patients in Life Island the problems and limits of this therapy method are represented. Microbiological findings are described in detail and commented. Sterile care is an essential part of an effective therapy of haemoblastoses, malignant lymphoma, and some solid tumours, particularly in the aplastic phase.


[Studies on the manufacture of hematological and cytological specimens using the centrifugation technic]. Centrifugal preparation (Cp) represents a method of creating haematological parameters which, in addition to a qualitative improvement of representing single cells, is suitable to produce reproducible haematological smear preparations. In qualitative and quantitative respect centrifugal preparations will reflect the intravasal conditions better than the common blood smear technique. Further advantages are the concentration of cellular elements and in connection with it the opportunity of finding cells rarely present. With a small amount of work centrifugal preparations enable counting values to be established as well as cytological and cytochemical findings from the venous blood to be obtained. As a result of the test it turned out that the common glass slides cannot be replaced by plastic foils ones without certain disadvantages appearing. First investigations of attempting to represent cells in culture medium as well as analyzing liquors and processing other biological liquids with the help of the centrifugal preparation technique brought unsatisfactory results. Centrifugal preparation provides the possibility of performing pre-programmed haematological examinations beginning with blood collection and ending up in the evaluation by a computer.


The transient HLA antigenic loss dependent on hormonal state of organism in a healthy woman. In a healthy 44-year-old female blood donor with HLA phenotype HLA-A 2, A 10/B 7, BW 15 repeatedly the transient loss of HLA antigens was proved during the menstrual period and supposed ovulation. Within the interval between menstruation and ovulation the antigens could reliably be proved. The results support the view that in women hormonal effects may influence the HLA antigenic expressivity. The significance of this observation for reliable typing of HLA antigens in women is discussed.


[Anemia in chronic hemodialysis].

[Anemia in chronic hemodialysis]. On the basis of general haematological, clinicochemical, cytomorphological, cytochemical and nuclearmedical investigations the anaemia of chronic haemodialysis can be considered as a renal anaemia modified by a permanent dialysis. It is less characterized by lack of erythropoietin, but more by a strong lack of iron, moderate haemolysis and slight deficiency of folic acid. After gradual diagnostics the substitution treatment with iron is absolutely necessary, medication of folic acid should be attempted. A permanent therapy with 80-120 mg of iron weekly administered parenterally can be successfully performed and 10-15 mg of folic acid daily, which will markedly lower the rate of transfusion.


[The possibility of use of amidinated human erythrocytes as agglutinin adsorbents in hemagglutination tests]. This study is concerned with the agglutinability of amidinated human erythrocytes. Amidinated erythrocytes behave like an immuno-adsorbent. Following the adsorption of an agglutinin, amidinated erythrocytes agglutinate specifically with native red blood cells or with native eukaryotic cells, if the glycocalyx of these native cells is provided with the respective agglutinin receptor groupings.


[Hemagglutination with Gal-N-Ac receptors in the hepatocyte membrane]. By means of mixed agglutination of isolated rat hepatocytes with human group A or rat erythrocytes, both of them were previously trypsinized and sialolyzed, respectively, a hepatocytic N-acetylgalactosamine-receptor was demonstrated. Gal-N-ac specifically inhibits this agglutination. Following oxidation with periodic acid red blood cells no more agglutinate with rat hepatocytes. This agglutination is not related to proteins adsorbed to hepatocytes. The agglutinability of erythrocytes and hepatocytes may bear some relevance to the elimination of old red blood cells.


Three cases of hemoglobin O Arab.

Three cases of hemoglobin O Arab. Three new cases of Hb 0 Arab in two families are reported from districts of Bulgaria, where a carrier state  of this abnormal hemoglobin has not been established so far. One of the propositi is a double heterozygote for Hb 0 Ar/beta(0)-thalassemia. His father is a simple heterozygote for Hb 0 Ar with clear-cut cytomorphological stigmata, indicating hemoglobinosis. The second propositus, according to clinical and laboratory data is also a double heterozygote for Hb 0 Ar and beta(0)-thalassemia. The carriers investigated are of Bulgarian nationality. Their territorial origin supportsthe thesis that the gene mutation for Hb 0 Arab most probably has taken place out of the present boundaries of Bulgaria.


[A-like specificity in Tn-activated erythrocytes of blood group B]. A Tn-polyagglutinability was observed in a 35 years old female patient with metrorrhagia. The A-like specificity of Tn-activated erythrocytes found simultaneously is especially significant, because this patient belonged to the blood group B, up till now, however, A-like findings have been described only for blood group O.


Glass activation-induced decrease or disappearance of normotest-thrombotest discrepancy in congenital coagulation disorders of the prothrombin complex. Exposure of plasma to glass for 60 minutes will cause a sharp, progressive shortening of Thrombotest clotting times in all congenital disorders of the prothrombin complex. On the other hand Normotest clotting times will change only slightly or moderately after the same glass activation. Consequently the Normotest-Thrombotest percentile discrepancy, quite marked at zero time, became progressively smaller. This behaviour is similar to that observed in a group of coumarin treated patients. These studies indicate that the Normotest/Thrombotest discrepancy does not allow any differentiation to be made between congenital coagulation disorders and coumarin induced defect. This is consistent with the non-specificity of the phenomenon.


[Prestenotic narrowing in the upper urinary tract (author's tract)]. While prestenotic dilatation in the upper urinary tract is considered as a characteristic radiological feature, earlier narrowing due to compensating hypertrophy of the musculature is either ignored or misinterpreted. Several examples of prestenotic narrowing are demonstrated and their significance in radiological diagnosis is stressed. This raises the question whether the narrowing of the ureters, seen in cases of bladder neck obstruction, particularly with prostatic hypertrophy, can be explained by a similar physiopathological response. This question can only be answered by performing pressure measurements during the course of radiological examination; the results would, however, be of doubtful value since the introduction of the ureteric catheter would lead to an additional unphysiological state.


[Controlled study on the efficacy of external treatment in surface-near thrombophlebitis]. The efficacy of an organo-heparinoid ointment (Hirudoid 40,000) compared with a placebo ointment was tested in a double blind trial on 40 patients with post-operative infusion thrombophlebitis. It was possible to provide objective data by means of the 125I-fibrinogen test for accelerated regression of thrombi following treatment with organo-heparinoid. The differences found between the 2 treatment groups were statistically significant. The clinical results conformed largely to the measured data. Especially pain and swelling were favourably affected by treatment with the active compound. 1 case of local allergic reaction to organo-heparinoid was observed. No further side-effects were seen.


Comparative binding specificity of methyltrienolone in human and rat prostate. The binding of methyltrienolone (R 1881) in crude human hyperplastic prostate cytosol was determined by a charcoal assay. Maximum binding was observed after 2-3 h of incubation at 0 degrees C. This binding decreased steadily thereafter and reached 41% of the 2-hour values after 96 h of incubation. In human hyperplastic prostate, the binding of 3H-R 1881 was competed by low concentrations of R 1881, R 5020 and progesterone and by high concentrations of dihydrotestosterone (DHT) and 17 alpha-methyl-DHT. In the rat prostate, on the other hand, this binding was competed by low concentrations of DHT and 17 alpha-methyl-DHT and only by high concentrations of progesterone and R 5020. The apparent association constant (Ka) of R 1881 was determined in three human prostates and found to be 0.2-0.4 X 10(9) liters/mol; the number of binding sites ranged from 101 to 158 fmol per mg of protein. These findings constitute further evidence for the existence of relatively large amounts of a progesterone-binding component in human hyperplastic prostate.


[Isolation and characterization of an unknown, leucine-rich 3.1-S-alpha2-glycoprotein from human serum (author's transl)]. This article describes the isolation and characterization of a previously unknown, leucine-rich 3.1S-alpha2-glycoprotein from human serum. The starting material was Supernatant II, which is a byproduct in the large-scale preparation of albumin and gamma-globulin by the ethacridine lactate/ammonium sulfate procedure. The purified protein is homogenous both in carrier-free and molecular-sieve electrophoresis. Its electrophoretic mobility indicates that it belongs to the alpha2-globulins. Isoelectric focussing splits it into 4 bands with isoelectric points between 3.8 and 4.1. In the ultracentrifuge it sediments in a single band at 3.1S. The molecular weight determined by equilibrium sedimentation is 49 600 +/- 4 000. Subunits were not detected. Chemical analysis reveals it to be a glycoprotein with a carbohydrate content of 23%. The amino acid content is unusual in that the leucine content is almost 17%, i.e. about every fifth amino acid is a leucine. The average concentration of the leucine-rich 3.1S-alpha2-glycoprotein in human serum was determined by a quantitative immunological method as 2.1 mg per 100 ml. The protein is not related to any of the previously known well characterized serum proteins.


Localization chromatid breaks in Fanconi's anemia, using three consecutive stains. The location of 339 break points was analyzed in three patients with Fanconi's anemia, using three consecutive stains: ordinary Giemsa, Q-banding, and R-banding. Almost all the breaks seem to take place in the Q bands, using R-banding, and in the R bands, using Q-banding. A very important artifact, varying according to the method used, is thus demonstrated. In fact, the breaks take place in the interbands, between R and Q bands. The breaks were also localized in relation to sister chromatid exchanges (SCEs), seen after BUDR treatment. There is a clear excess of breaks at place of SCE (29%). This may indicate a possible correlation between breaks and SCEs.


Frequency of Ag-stained nucleolus organizer regions in the acrocentric chromosomes of man. The Ag-stainability of the nucleolus organizer region (NOR) was studied in the acrocentric chromosomes identified by Q-banding of cultured lymphocytes in 51 karyotypically normal persons (31 males and 20 females). A consistent pattern of Ag-positive NORs was found in each individual. Ninety percent of individuals have a model number of 8--10 Ag-positive NORs per cell. The frequency of Ag-positive NORs is similar in all five acrocentrics. A statistically nonsignificant lower frequency is found in chromosome 22. Ag-negative NORs on both homologues were found in four cases. The observed frequency distribution of individuals with homozygous NOR-positive, heterozygous, and homozygous negative acrocentric chromosomes was in accordance with the Hardy-Weinberg law in all five pairs of the acrocentric chromosomes as well as in total. No sex difference was observed on our material.


Combined silver staining of the nucleolus organizing regions and Giemsa banding in human chromosomes. A combination of the silver-staining method of the nucleolus organizer regions (NORs) with a Giemsa-banding method is described. This double staining allows a rapid identification of the NOR-bearing chromosomes.


Hyperacute allergic encephalomyelitis: a single determinant. HEAE cannot be induced in either guinea pigs using 10(10) or 20x10(10) organisms of B. pertussis or in rabbits using 4x10(10) organisms of B. pertussis and basic protein from the following species: guinea pig, Lewis rat, human, bovine, porcine, monkey and rabbit. The only encephalitogenic region for Lewis rats which also produces HEAE in Lewis rats is not encephalitogenic in either rabbits or guinea pigs. Therefore it seems highly probably that there is only one HEAE determinant for all species which are able to express HEAE.


Tolerance induction as an index of age-related changes. Tolerance to rabbit gamma globulin RGG) can be induced in newborn or three week old mice of all strains tested. In six week old SJL or NZB mice there is marked resistance against tolerance induction to RGG. This resistance increases with increasing age and is completed by the age of 12 weeks. Resistance to tolerance can be observed in 3 week old SJL animals when human IgG3 but not when human IgG1 or RGG is employed as tolerogen. Using IgG3, resistance to tolerance induction can be demonstrated in 28-34 week old A/J, C57BL/6J and DBA1/J mice. Thus resistance to tolerance induction seems to be a concomitant of aging; SJL and NZB differ from other strains only by the age at which this change is apparent. By the 6th week of age, SJL and NZB mice develop marked resistance to tolerance induction with RGG, and so do hybrids between these two strains; there is no complementation with respect to resistance against tolerance induction. Aggregate-freed iodinated rabbit gamma globulin (125I-RGG) but not mouse immunoglobulin is eliminated much more rapidly from the body of normal 6 week old SJL and NZB mice than from the body of hybrids. Complementation in the hybrids might occur and prevent a response to small quantities of aggregate-freed RGG.


Arming of macrophages with lymphocytes from mice immunized with tumor tissue. C57BL/6 macrophages are rendered specifically cytotoxic ("armed") by incubation with spleen cells from C57BL/6 mice sensitized in vivo with allogeneic P-815 mastocytoma cells. Data are presented which suggest that the cytotoxic activity of the macrophages is not due to the adherence of sensitized lymphocytes. The results indicate that it is the T-lymphocyte which is primarily involved in arming. Furthermore it is the cytotoxic and not the proliferating T-cell which is responsible for arming.


Studies of sperm antigenicity. 6. In vivo and in vitro cellular reactivity in guinea pigs sensitized with fractions of guinea pig spermatozoa. Normal guinea pig spermatozoa cells were homogenized by a French pressure cell. Three soluble and three insoluble fractions were obtained by ultrascentrifugation and (emulsified in CFA) were used for guinea pig sensitization. The following were observed: 1) all fractions were immunogenic except one; 2) in vivo and in vitro delayed hypersensitivity was elicited in animals immunized with these fractions; 3) two distinctive histopathologic lesions were observed in the testes of sensitized animals: lesions of orchitis type developed in animals injected with some fractions. Other fractions induced lesions of aspermatogenic type. These results correlated well with delayed hypersensitivity results obtained by in vivo and in vitro tests. Although some other spermatozoal fractions did not cause severe changes in the testes. The lack of sperm accumulation in the epididymis was obvious.


Antigenicity of lateral flagella of Vibrio parahaemolyticus: existence of two distinct antigenic determinants in a flagellum. Cells of Vibrio parahaemolyticus treated with Formalin agglutinated with anti-lateral flagella antiserum. On the basis of agglutination tests, antigens of lateral flagella were divided into three groups. HL1, HL2, and HL3. However, in gel diffusion tests, flagellins prepared from strains belonging to different groups showed a common antigenicity. It is assumed that these results are due to the existence of two distinct antigenic determinants in the lateral flagella. One of them exists on the surfaces of the flagella and is responsible for H-agglutination, and the other exists inside the flagella and is exposed when the flagella are solubilized to flagellin monomers.


Immunization with R mutants of Salmonella minnesota. II. Serological response to lipid A and the lipopolysaccharide of Re mutants. Rabbits were immunized with Salmonella minnesota Re mutant bacilli or lipid A, and the serological responses were compared. Re lipopolysaccharide and lipid A were found to possess distinct antigenic determinants as measured by hemagglutination, hemolysis, and precipitin techniques. Lipid A preparations from several enterobacterial and non-enterobacterial species were shown to cross-react in agar gel precipitation reactions. In contrast, preparations from bacilli that do not contain glucosamine as an integral part of their lipid A structure did not demonstrate cross-reactivity with enterobacterial lipid A. Antibody to smooth heterologous bacilli present in some antisera prepared against Re bacilli or lipid A was directed specifically against the smooth lipopolysaccharide and not against the Re or lipid A determinants, demonstrating a nonspecific mitogenic response. Precipitation and immunofluorescence tests indicated that the Re determinant was available on smooth bacilli for reactions with specific antisera but that the lipid A determinant was not.


Surface immunoglobulin on rabbit lymphoid cells. V. Ultrastructural distribution of b4 allotypic determinants of thymus, lymph node and bone marrow lymphocytes. An indirect immunoferritin-labeling technique is used to localize cell surface immunoglobulin (Ig) allotypic determinants on rabbit lymph node, bone marrow and thymus lymphocytes. 47% of the lymph node cells, 62% of bone marrow small lymphocytes and less than 1% of thymic lymphocytes, are positive for surface Ig. Two Ig-positive lymph node lymphocyte populations which differ in fine structure are identified but both cell types express similar amounts of surface Ig (7,000-28,000 Ig molecules per cell). Bone marrow small lymphocytes make up only 9% of the total cell population and express less surface Ig (ca. 3,000-12,000 molecules Ig per cell) than most Ig-bearre described and both are essentially negative for surface Ig using this labeling technique.


Further studies on passive peritoneal anaphylaxis in the rat. Four compounds with H1 anti-histamine activity and four adrenoceptor stimulants, each given to rats prior to passive peritoneal anaphylaxis (PPA), inhibited extravasation of serum proteins into the peritoneal fluid at doses which had no effect on histamine release. In contrast, aminophylline and some non-steroidal anti-inflammatory agents inhibited extravasation only at doses which inhibited histamine releases they showed a similar type of avtivity to that of disodium cromoglycate (DSCG) and a nitroindanedione (BRL 10833), although they were much less potent. Predosing with DSCG reduced the potency of subsequent doses of DSCG, BRL 10833 and indomethacin, but not of aminophylline or phenylbutazone, and therefore DSCG, BRL 10833 and indomethacin may share a common pathway to produce activity. In the rat PPA system, no evidence was found for histamine 'feedback' inhibition of histamine release.


Studies of the nature of extracellular components of rat seminiferous tubular wall. II. Immunological characterization of basement membrane antigen. An antiserum was obtained in rabbits using as antigen a preparation rich in basement membranes isolated from normal rat testis. By passive hemagglutination test a titer of 1/20,000 was obtained when the antiserum was reacted with the specific antigen; no cross reactions were detected with other fractions extracted from the germinal cells of rat testis. A weak cross reaction was obtained when the antiserum was reacted with collagen extracted from rat testis. Moreover, the antiserum cross reacted with other isolated basement membranes such as glomerular basement membrane of rat kidney and sheep lens capsule. By the indirect immunofluorescent technique the localization of the antigen antibody reaction was detected at the seminiferous tubular wall and vessels. No reaction was observed in other structures of the testis except for a faint reaction at the interstitial collagen fibers. Both, serological and immunohistochemical techniques demonstrated: (1) the existence of common antigenic determinants among basement membranes of different organs of the rat, (2) a partial immunological identity among basement membranes of different species.


[Mechanism of mediators in human skin. Pharmacoanalysis and therapeutic aspects]. Under physiological and pathological conditions, the skin is influenced by tissue hormones (biogenic amines, kinines, prostaglandines), neuro-hormones (acetylcholine, catecholamines) and by proteins as complement. The combination of neurological stimulation with a series of self-propagating metabolic changes, shows new aspects of tissue reaction. Histamine triggers a serie of mediators as kinines and prostaglandines. Due to their influence on mediators and their effects on regulatory mechanisms the cyclic nucleotides can serve as model substances for the development of therapeutic processes for dermatological as well as for other disorders. The influence on mediator-systems by topical treatment and proven permeability of such substances is of extreme importance for the future development of local dermatological therapy.


Demonstration of intracytoplasmic glycogen of megakaryocytes and blood platelets by means of the periodic acid-thiocarbohydrazide-silver proteinate method. The glycogen of megakaryocytes and blood platelets has been investigated in glutaraldehyde and osmium tetroxide fixed tissues by the periodic acid-thiocarbohydrozide-silver proteinate method (PA-TCH-SP). The PA-TCH-SP method involves the staining of intracytoplasmic glycogens more densely than the routine lead citrate method. Glycogen having a mean particle diameter of 21.1 nm has been shown localizing in the matrix of mature megakaryocytes, while that of glycogen in the platelets was 26.2 nm. The staining pattern of the glycogen in blood platelets was classified into three groups according to staining intensity. It is found that the PA-TCH-SP method is a very suitable one for the demonstration of intracytoplasmic glycogen from the viewpoints of reaction specificity, reproducibility, fineness of reaction products, sufficiency of electron density, and experimental cost. This method is also a very useful one for differentiating intracytoplasmic glycogens and ribosomes.


Mouse epidermal growth factor: light and electron microscopical localisation by immunocytochemical staining. Epidermal Growth Factor (EGF) has been localised by immuno-staining to granules of the convoluted duct cells of the submaxillary glands of mice. Improved techniques of freeze drying and formaldehyde vapour fixation have resulted in a light microscopical localisation sharper than was achieved by previous methods. EGF has also been identified by electron immunocytochemistry using the unlabelled antibody enzyme method. EGF is present in greater quantities in male mice than in female mice but in pregnant females the level of EGF in the submaxillary gland is equal to that of the male. It declines gradually during the three weeks of lactation. In view of the chemical similarity between mouse EGF and human Urogastrone these improved methods of identification may be useful in the localisation of the human substance.


Silver staining the caveolae intracellularis of smooth muscle. Bodian silver staining of guinea-pig Taenia coli produces a regular distribution of silver at the periphery of the smooth muscle cells. Electron microscopic analysis of this precipitate revealed that the membrane of the caveolae intracellularis was the only structure at the periphery of the cells that consistently exhibited a high affinity for silver. Thus, this staining method demonstrates that caveolae are distributed over most of the cell surface in an orderly pattern.


Chemistry of bleomycin. XVIII. carbon-13 NMR studies. During chemical studies of bleomycin, many fragments and derivatives have been isolated and characterized. The 13C-NMR spectra of these compounds were taken and analyzed for structural information, and the complete assignment of the spectra was achieved. The 13C-chemical shift map thus obtained contains information about the structure and conformation and will be useful for studies on the chemistry and biology of bleomycin and related compounds.


Biochemical studies on an acidophilic, thermophilic bacterium, Bacillus acidocaldarius: isolation of bacteria, intracellular pH, and stabilities of biopolymers. Acidophilic, thermophilic bacteria were isolated from Japanese acidic hot springs. They were spore-forming rods, identified as Bacillus acidocaldarius. DNA extracted from these acido-thermophiles showed no abnormality in chemical structure; it was instantly denatured and gradually decomposed giving rise to apurinic acid in a hot acid environment milder than the optimal conditions for the growth of the acido-thermophiles. Glyceraldehyde-3-phosphate dehydrogenase extracted from B. acidocaldarius was not active at pH 5 or less, and was resistant to heat at neutral but not acid pH. The intracellular pH was computed to be neutral by using dimethyl-2,4-oxazolidinedione. When uncouplers or inhibitors of respiration were added to the cells suspended in hot acid solution, the estimated pH was not changed and glyceraldehyde-3-phosphate dehydrogenase in the cells was not denatured. These results suggest that the cytoplasm of B. acidocaldarius is a hot neutral environment, and that a pH gradient across the cell envelope can be maintained even when oxidative phosphorylation or respiration is inhibited.


Effect of Rauscher leukemia virus-specific proteins on reverse transcriptase. Binding between reverse transcriptase and p30. The reverse transcriptase was purified to homogeneity from Rauscher leukemia virus by sequential column chromatography on phosphocellulose and DNA-cellulose. The purified enzyme, a single polypeptide chain with a molecular weight of approximately 70,000, interacts with major internal protein p30 of the same virus. The reverse transcriptase - p30 complex stimulated [3H]TMP incorporation into (dT)12 - (rA)n 2- to 3-fold compared to that observed with the purified enzyme alone. Monospecific antiserum made against either p30 or reverse transcriptase precipitated the entire complex. The sedimentation rate of the reverse transcriptase - p30 complex is approximately 12 S as estimated by glycerol gradient centrifugation, and the molecular weight is approximately 400,000 by chromatography on a Sepharose 6B column. The complex dissociates into its original components when treated with 0.8 M KCl.


Selective radioactive labeling of cell surface sialoglycoproteins by periodate-tritiated borohydride. Low concentrations of sodium metaperiodate induce specific oxidative cleavage of sialic acids between carbon 7 and carbon 8 or carbon 8 and carbon 9. The aldehydes formed can easily be reduced with NaB3H4 to tritiated 5-acetamido-3,5-dideoxy-L-arabino-2-heptulosonic acid or 5-acetamido-3,5-dideoxy-L-arabino-2-octulosonic acid. At 0 degrees, the periodate anion penetrates the cell plasma membrane very slowly and only externally exposed sialic acids are oxidized. This was shown by (a) limited labeling of the sialoglycoproteins in a preparation of inside-out erythrocyte vesicles; (b) trapping 14C-labeled fetuin within resealed erythrocyte ghosts; fetuin was then poorly labeled, whereas the erythrocyte sialoglycoproteins were highly labeled; (c) comparison of labeled glycoproteins of mouse lymphoid cells before and after treatment with neuraminidase. This simple method of specifically introducing a radioactive label into cell surface sialic acids is useful in the study of cell surface sialic acid-containing glycoproteins.


Antigenic similarity of heat-labile enterotoxins from diverse strains of Escherichia coli. With use of the rabbit intestinal loop model, heat-labile enterotoxins from 21 Escherichia coli strains isolated from a wide spectrum of patients with diarrheal diseases were all neutralized to high titer by two antisera prepared against enterotoxins of either E. coli or Vibrio cholerae. These findings suggest marked immunological similarity among heat-labile enterotoxins from a heterogenous group of E. coli.


Purification of equine infectious anemia virus antigen by affinity chromatography. Affinity chromatography was performed to obtain highly purified antigen from equine infectious anemia (EIA) virus. After crude antigen was concentrated by polyethylene glycol precipitation of culture fluids from equine dermal cells persistently infected with EIA virus, and after the virus was disrupted with ether, it was added to a column of cyanogen bromide-activated Sepharose 4B to which EIA-specific antibody had been conjugated. The antigen was effectively released from the column with 5M MgCl2 and proved to be highly purified. Passive hemagglutination tests on sera from EAI infections were carried out, using the purified antigen. Results indicated that the passive hemagglutination test with the antigen was a specific laboratory test with high sensitivity for EIA infection.


Plaque assay and cloning of scrub typhus rickettsiae in irradiated L-929 cells. It was demonstrated that gamma-irradiated L-929 cells support plaque formation by three strains of Rickettsia tsutsugamushi and representative species of the spotted fever and typhus group rickettsiae. Sensitivity of the plaque assay for detection of viable scrub typhus rickettsiae was similar to that achieved with intraperitoneal inoculation of random-bred mice. The concentration of irradiated cells and the temperature and length of incubation were all found to affect plaque size. A technique combining terminal dilution and plaque purification was used to obtain clones of three strains of scrub typhus rickettsiae.