Mutagenic and lethal effects of alpha-benzene hexachloride, dibutyl phthalate and trichloroethylene in Saccharomyces cerevisiae. Saccharomyces cerevisiae strain XV185-14C for reversion studies was used to investigate the genetic activity of alpha-benzene hexachloride dibutyl phthalate and trichloroethylene. The results indicate that none of the three compounds was genetically active when yeast cells were treated in phosphate buffer (pH 7.0) in the absence of metabolic conversion. However, in the presence of the 9000 g supernatant of mice liver homogenate, NADP, glucose-6-phosphate, phosphate buffer (PH 7.4), MgCl2, KCl, the components which were used for the metabolic conversion, trichloroethylene porved to be a powerful mutagen. It increases the frequency of homoserine, histidine and lysine revertants over those of the control levels. Trichloroethylene appears to induce frameshift as well as base substitution mutations.


Short-term antithyroid drug therapy for the thyrotoxicosis of Graves's disease. We investigated whether thyrotoxic patients treated with short-term antithyroid therapy would achieve prolonged remissions. Thirty-one previously untreated and nine previously treated patients with thyrotoxic Graves's disease received a single daily dose of methimazole or propylthiouracil. The drug was stopped at, or shortly after, the time they became euthyroid. Twelve of the 31 previously untreated patients remained in remission for 29 +/- 3.5 months (mean +/- S.E.) after treatment for 4.5 +/- 0.3 months. Four of the nine previously treated have remained in remission of 13.0 +/- 2.1 months after treatment for 3.0 +/- 0.3 months. Of various possibilities analyzed, only a small goiter at the onset of therapy and tri-iodothyronine toxicosis were significantly favorable prognostic indicators that a remission would be maintained. The lasting remission rate is as good when antithyroid drugs are stopped as soon as the patient is euthyroid as when they are continued for one year or more.


Clonidine-induced locomotor hyperactivity in rats. The role of central postsynaptic alpha-adrenoceptors. The alpha-adrenergic agonist, clonidine, causes sedation in normal rats. The present study demonstrates that clonidine evokes strong locomotor stimulation in rats pretreated with 6-hydroxydopamine plus reserpine. Similar, but less intensive hyperactivity is observed in rats given clonidine after combined pretreatment with 6-hydroxydopamine plus p-chlorophenylalanine plus alpha-methyl-p-tyrosine, or with reserpine plus low doses of yohimbine. The aplha-adrenolytic drugs, phenoxybenzamine, phentolamine and aceperone, as well as high doses of yohimbine, antagonise the clonidine-induced locomotor stimulation; in contrast, the dopamine receptor blocking agents, pimozide and spiroperiodol, exert no antagonistic effect. The results indicate that in the brain of normal animals, clonidine predominantly activates presynaptic alpha-adrenoceptors on noradrenergic neurones and thereby induces sedation. After destruction of the noradrenergic fibres by 6-hydroxydopamine plus reserpine, activation of postsynaptic alpha-adrenoceptors prevails so that hyperactivity results.


Mechanism of action of neurohypophyseal hormones on sodium excretion and diuresis. Administration of pituitrin, vasopressin, and oxytocin to rats during spontaneous micturition increases diuresis and sodium excretion by reducing tubular reabsorption. In adrenalectomized rats these preparations have no diuretic effect and increased sodium excretion is observed only after administration of vasopressin. After hypophysectomy the diuretic effect of the preparations disappears but they still increase sodium excretion. It is postulated that the diuretic effect of the neurohypophyseal hormones is connected with activation of the pituitary-adrenal system, whereas some additional mechanism is involved in their effect on sodium excretion.


[Cellular bases of immunological recognition. The precursors of antigen-binding T- and B-lymphocytes and the patterns of their maturation]. The peculiarities of carefully studied immunoglobulin receptors of the specialized clones of B-lymphocytes are described. These latter are precursors of antibody-forming cells but play only a limited role in the recognition of foreign antigens which is realized by T-lymphocytes. The differentiation of T- and B-lymphocytes from the single precursor, a bone marrow stem hemopoietic cell, proceeds in the bone marrow itself, thymus and peripheral lymphoid organs in several discrete stages. At each of these stages the cells have definite properties and, in some cases, may be separated one from another. The modern data are given concerning the properties of cells at every stage of antigen-independent differentiation of T- and B-lymphocytes which proceeds during embryonic and postnatal development, its alternative pathways, factors of regulation, role of local microenvironment and possibilities of modelling in vitro.


[Immunohistochemical study of alpha-fetoprotein in the postnatal ontogeny of mice with a varying sensitivity to liver carcinogens]. The ontogenesis of alpha-fetoprotein (AFP) was studied in the C57BL, CC57BR, C3H/He, A/He and DD mice during the first 4 weeks of life. The AFP hepatocytes were determined by indirect immunofluorescent staining of liver cells isolated with sodium tetraphenyl borate; the content of AFP in blood was determined by the method of rado immunodiffusion. The methods utilized allowed to obtain the quantitative characteristics of the dynamics of AFP-positive cells in the liver and the content of AFP in the blood. In the newborn mice over 90% hepatocytes contain AFP, the intensity of luminescence being heterogenous. The number of bright-liminescent cells equals to 50% during the first day of life rapidly decreases and none of them are found by 9--11 days. The number of average- and weak-luminescent hepatocytes does not decrease during the first 10 days, but then gradually decreases and none of them are found by 23 days in the CC57BR and by 27 days in the DD and A/He mice. A comparison of the dynamics of AFP-positive cells in the liver and the content of AFP in the blood has shown that the bright-luminescent hepatocytes are the main producent of this protein in the early postnatal ontogenesis.


Fine particles of cotton dust influence histamine release. Cotton dust particle distribution in respiratory airways was measured to determine if specific size particles of the heterogeneous dust are concentrated as a function of airway diameter. Previously reported work indicated that the fine particles are responsible for the decrement in pulmonary function which occurs subsequent to cotton dust inhalation. After recovery, particles were sized and counted microscopically. Significant increases in fine particles (5-7 mu) were found in all airways of exposed animals as compared with controls. These particles become more concentrated as the airway diameter decreased. Histamine determinations were made in mice lungs after the animals were exposed to cotton dust for various periods of time to see if such treatment could be correlated with elevations in the concentrations of this biogenic amine. Lung histamine increased as exposure time to cotton dust increased. It is concluded that the lung histamine release occurring after cotton dust inhalation is related to some property of the fine particles of the heterogeneous dust.


Trigger points and acupuncture points for pain: correlations and implications. Trigger points associated with myofascial and visceral pains often lie within the areas of referred pain but many are located at a distance from them. Furthermore, brief, intense stimulation of trigger points frequently produces prolonged relief of pain. These properties of trigger points--their widespread distribution and the pain relief produced by stimulating them--resemble those of acupuncture points for the relief of pain. The purpose of this study was to determine the correlation between trigger points and acupuncture points for pain on the basis of two criteria: spatial distribution and the associated pain pattern. A remarkably high degree (71%) of correspondence was found. This close correlation suggests that trigger points and acupuncture points for pain, though discovered independently and labeled differently, represent the same phenomenon and can be explained in terms of the same underlying neural mechanisms. The mechanisms that play a role in the genesis of trigger points and possible underlying neural processes are discussed.


Acupuncture and chronic pain mechanisms: the moderating effects of affect, personality, and stress on response to treatment. The present study, part of a larger project investigating neurophysiological and psychosocial factors affecting response to acupuncture for chronic pain, compares responders and non-responders to acupuncture on a series of variables assessing personality, affect and stress. Subjects were 40 patients with pain beneath the waist level longer than 6 months duration selected from the roles of the Multidisciplinary Pain Clinic. Responders, defined as 50% or more reduction in pain estimate for greater than two weeks, were found to be less depressed, less passive and overly conventional, have shorter duration of pain, endorse less frequent exposure to stressors, and have less serious non pain-related illnesses. The findings are viewed as linking the intractability of pain states with psychosocial factors which may interfere with response to somatic modes of therapy or which may interfere via alterations of tonic neurohumoral factors. The study also in seen as supporting the importance of considering psychological variables in evaluating patients for pain treatment strategies and suggests inclusion of such variables in investigating response to other modalities of treatment for chronic pain.


Choice of strong analgesic in terminal cancer: diamorphine or morphine? A controlled trial of diamorphine (diacetylmorphine, heroin) and morphine is reported in which the two drugs were administered regularly by mouth in individually determined effective analgesic doses. Elixirs contained cocaine hydrochloride 10 mg/dose; other drugs were prescribed when indicated clinically. 699 patients entered the trial and, of these, 146 crossed from diamorphine to morphine, or vice versa, after about two weeks using an oral potency ratio of 1.5/1 determined in a pilot trial. Additional medication and survival were closely similar in both treatment groups. In the female crossover patients, no difference was noted in relation either to pain or the other symptoms evaluated. On the other hand, male crossover patients experienced more pain, and were more depressed, while receiving diamorphine. In these, the potency ratio of diamorphine to morphine appeared to be less than 1.5/1. If this is allowed for, then the difference in mood is probably not significant. Compared with male patients, fewer females required a dose of 10 mg or more, but more were prescribed an anxiolytic. The ability to do without a 2 a.m. dose appeared to be related more to the size of the dose than to gender or treatment. It is concluded that, provided allowance is made for the difference in potency, morphine is a satisfactory substitute for orally administered diamorphine. However, when injections are necessary, the greater solubility of its hydrochloride gives diamorphine an important practical advantage over morphine, especially when large doses are required.


[Immunochemical study of the lipoprotein substance Pg (author's transl)]. Site Pg is an antigenic determinant which is present on the surface of serum lipoproteins and reacts with an antilpoprotein myelomatous immunoglobulin, the IgA, Ger. Firstly site Pg is extracted from LDL with the major part of lipids, by an ether-methanol mixture at 6 p. 100. Then the organic substrate obtained is evaporated under vacuum and dissolved in saline before being submitted to an extraction by ehter only. Under these conditions, site Pg remains in the aqueous phase. A polyacrylamide gel filtration on Biogel P2 allows then to separate the substance which reacts with IgA Ger. anti-Pg, from the aqueous phase. The identification of site Pg allowed us to recognize the presence of galactose, phosphorus and choline, and to evaluate its molecular weight of about 330. The reactivity of site Pg with IgA anti-Pg was measured by inhibition of passive hemagglutination and fluorescence quenching. The association constant of site Pg with the whole IgA Ger. or with its Fab fragment could thus be calculated (1.6 10(5) M-1 with the whole IgA; 2.3 10(5) M-1 with the Fab fragment). We showed also that if phosphorylcholine plays an immunodominant role, the presence of sugar seems necessary, since the association constant of the whole IgA Ger. with site Pg is higher than that of IgA Ger. with phosphorylcholine alone.


An experimental evaluation of staining techniques for the detection of early ischaemic injury to the myocardium. A series of experimental infarcts of the posterior papillary muscle of the canine heart was used to assess the value of 6 special stains in the histological detection of early myocardial infarction. The infarcts were of 5-720 min duration and were compared not only with normal control myocardium but also with normal myocardium autolyzed for similar periods of time. All tissue was stained with H & E, PAS, PAS-diastase, PTAH, Masson's trichrome, Connor's modification of the acid fuchsin method, Puchtler's PAS-navy blue, and Lie's haematoxylin-basic fuchsin-picric acid. The zone of severely altered myofibres which separated normal from ischaemic tissue in infarcts aged 2 or more hours was demonstrated by all but PAS. Normal, border, and ischaemic zones of heart muscle were clearly differentiated only by PTAH and PAS-navy blue. PAS distinguished normal from glycogen-depleted ischaemic myocardium after only 40-60 min, but this change was also seen in autolyzed tissue.


Prognostic importance of ventricular extrasystoles in acute myocardial infarction. Ventricular arrhythmias were recorded in 233 patients in a prospective study of patients with acute myocardial infarction. In over 95% of patients antiarrhythmic therapy was not given until the onset of ventricular tachycardia, ventricular fibrillation, or persistent idioventricular  rhythm. There was a mortality of 18% during the patients' stay in hospital. The most important features of ventricular ectopic activity, which preceded these severe ventricular arrhythmias in the first 48 hr, were multiformity, variation of coupling intervals of larger or equal to 0-1 sec, the R-on-T phenomenon, double ventricular extrasystoles and ventricular bigeminy. The number of a single ventricular extrasystoles per minute was related to the probability of these severe ventricular arrhythmias but to a lesser degree. It was found that if all the patients with the first two prognostic features that if all the patients with the first two prognostic features were removed, the number of single ventricular extrasystoles was not of significant import and the other features were less important. Three-quarters of the severe arrhythmias occurred in the first 24 hr and during this period 60% were preceded by either multiform ventricular extrasystoles or extrasystoles with variable coupling. The importance of these findings in relation to prophylactic therapy is discussed.


[Antithyroid activity of various newly synthesized 1,3-thiazine derivatives]. Experiments were performed on rats. It was revealed that three newly-synthesized 1,3-thiazine derivative (1,3-thiazandithion-2,4, 1,3-thiazanthion-2-on-4-thiosemicarbazone-4, and 1,3-thiazandion-2,4) possessed and antithyroidal action in doses equal to 15% of LD50; in the two former preparations it was accompanied by an goiterogenic effect. 1,3-thiazandion-2,4 had the most antithyroidal action and relatively weak goiterogenic effect.


Plasma volume changes after infusion of various plasma expanders. In the immediate post-operative period after moderate surgical procedures, 1 litre of a colloid solution or saline was given intravenously. The plasma volume expansion after infusion of dextran 70 (Macrodex), hydroxyethylstarch (Volex), polygelatin (Haemaccel), albumin and saline was found to be between 790 and 180 ml. The most efficent plasma expander was dextran, followed by hydroxyethylstarch. Polygelatin and saline did not give full restitution, although twice the calculated loss was infused. Total plasma protein concentration was lowered in all groups in proportion to the dilution, except for the patients given albumin, in whom the concentration of total protein increased. Calculation of the total circulating protein mass showed no decrease during the period immediately after the infusion. This investigation has demonstrated that the most efficient plasma volume expander is dextran but that hydroxyethylstarch offers an almost equal alternative in terms of volume expansion. Dextran, however, exerts an advantageous effect on the microcirculation. As the metabolic pathways of hydroxyethylstarch have not yet been further explored, dextran is preferred when using artificial colloids. Judged by its secondary effects alone, including the influence on plasma protein patterns, albumin seems to be the compound of choice. Polygelatin and saline are not efficient expanders when hypovolaemia is to be corrected rapidly.


Circulation of H+ and K+ across the plasma membrane is not obligatory for bacterial growth. Streptococcus faecalis grows normally in the presence of gramicidin and other ionophores under conditions such that there is no gradient of pH or of electrical potential across the plasma membrane and that currents of H+, K+, and Na+ are short-circuited. Growth requires a rich medium, a slightly alkaline pH, and a high concentration of external K+. The proton circulation maintains the cytoplasmic pH and pools of ions and other metabolites but is not obligatory for biosynthetic functions including DNA replication, cell division, or assembly of the structural framework of the cell.


[Combination therapy of the undifferentiated supratentorial astrocytoma. Review of current possibilities of treatment]. A review is presented of the results published in the most important recent literature on the treatment of high grade supratentorial astrocytoma by surgery, surgery and radiotherapy, chemotherapy, and adjuvant chemotherapy after surgery and radiotherapy. The results of the different modalities are compared, the treatments of proven significance are described and future possibilities are discussed.


Kepone-induced scoliosis and its histological consequences in fish. Scoliosis in fish is caused by several diverse agents that possibly act on the central nervous system, neuromuscular junctions, or ionic metabolism. The organochlorine pesticide Kepone induces scoliosis in the sheepshead minnow. Some effects associated with Kepone-induced scoliosis in these fish are disruption of myotomal patterns, inter- and intramuscular hemorrhage, fractured centra of vertebrae, and death. The histological syndrome of Kepone poisoning in fish and the clinical syndrome in humans suggest that the nervous system is a primary target for Kepone and that scoliosis is a secondary effect of Kepone poisoning in fish.


Frequency of HLA antigens in chronic myelocytic leukemia. Histocompatibility antigen (HLA) phenotypes of 34 patients with Ph1+ chronic myelogenous leukemia (CML) were evaluated for association with HLA antigens. Two control populations were compared to the CML patients: 142 normal volunteer platelet donors, and 160 normal donors of granulocyte transfusions. HLA typing was done by lymphocyte microcytotoxicity tests for nine antigens on sublocus A and 15 antigens on sublocus B. HLA-B7 and HLA-B12 were decreased in CML patients compared to both platelet and granulocyte donors. There was increased frequency of HLA-A3 in patients (41%) as compared to controls (25% and 33%); HLA-B5 - patients = 20%; controls 8% and 6%; and HLA-BW17 - patients = 17%; controls = 6% and 3% (P = 0.01). Median survival was 24+ months and independent of HLA. HLA-B5 and HLA-BW17 were significantly increased in patients with CML compared to two normal control populations. No increase in HLA-B8 was seen. Decreased frequency of HLA-B7 and B12 was noted. The significance of these differences is being evaluated.


Leukemia and prostatism.

Leukemia and prostatism. Benign prostatic hyperplasia associated with chronic lymphocytic leukemia is relatively rare. Needle biopsy is indicated to determine the possible benefits of radiation therapy as compared to surgery. Although overall mortality is high, transurethral resection (TUR) is well tolerated and may be undertaken without undue concern. An additional case is reported.


The present status of cytochemistry in the diagnosis of haematological malignancy. Newer concepts in the application of cytochemistry to the diagnosis of haematological malignancies are discussed. The underlying physiological principles and location of the various enzymes are indicated. Older stains, once thought to be diagnostic, have been shown to be nonspecific. Newer, more accurate stains are now available.


Human gene mapping using an X/autosome translocation. Human fibroblasts containing a translocation between the X chromosome and chromosome 15 were fused with the 6-thioguanine-resistant mouse cell line, IR. Resulting hybrids, selected in HAT medium, retained the X/15 chromosome. Hybrids which were counterselected in 6-thioguanine lost this chromosome. The X-linked markers glucose-6-phosphate dehydrogenase (G6PD), phosphoglycerate kinase (PGK), and hypoxanthine phosphoribosyl transferase (HPRT), and the non-X-linked markers pyruvate kinase (PKM2) mannose phosphate isomerase (MPI), N-acetyl hexosaminidase A (HEXA) and beta2-microglubulin (beta2-m) all segregated in concordance with the X/15 translocation chromosome. The latter markers have been assigned to chromosome 15. Selection against the X/15 chromosome was done using antihuman beta2-m serum. Electrophoretic and immunochemical analyses of the N-acetyl hexosaminidases A and B in these hybrids were performed.


beta2-microglobulin locus on human chromosome 15. We have developed an autoradiographic/electrophoretic assay capable of distinguishing mouse and human beta2-microglobulin (beta2m) in spent culture media. The method is applicable to mouse and human lines and to hybrid cell lines made from them. With this technique, mouse/human hybrid cell lines were tested for the presence of human beta2m. Isozyme and karyotype analyses were also carried out with the hybrids. The combined results of these studies show that the structural gene for human beta2m is on the long arm of chromosome 15.


33258 Hoechst enhancement of the photosensitivity of bromodeoxyuridine-substituted cells. Cells that have incorporated 5-bromodeoxyuridine (BrdU) into their DNA become extremely sensitive to photoinduced killing after treatment with the dye, 33258 Hoechst. Under appropriate conditions the dye increases the efficiency of killing by a factor of approximately 10(4). This enhanced photosensitivity suggests that the combination of BrdU, dye, and light can be used to select conditional lethal mutants.


Clinical management of metastatic breast cancer. The physician treating the patient with recurrent or metastatic breast cancer has a number of available therapeutic options. Various clinical and laboratory factors play a role in the decision process for the choice of the initial and subsequent treatments for such patients. The strategies for palliative therapy are reviewed along with estimates of potential benefits of the various modalities.


Polygenic control of mixed lymphocyte reaction in dogs. The genetics of the MLR of the dog are studied using LD typing cells. The four new homozygous reference cells described bring the total number of defined LD specificities in this species to 11. In families, LD specificities generally segregate en block with the SD specificities of the DLA complex. The recombination frequency between the SD and LD loci is provisionally estimated to be 0.007. A slight but significant stimulation between two SD identical LD typed siblings could not be explained on the basis of a single recombination between the SD loci and one LD locus and therefore suggested the existence of an additional LD system. Unrelated SD different individuals with two different LD determinants in common frequently stimulate each other in MLR, but low responses are also encountered. The results described are compatible with a hypothesis of two segregant series of alleles for the genetic control of the MLR and linkage disequilibrium between certain alleles of these series. More conclusive evidence of the validity of this hypothesis and further knowledge of other factors which determine the response in the MLR are required before a reliable estimate of the importance of LD structures in histocompatibility can be made.


T-lymphocyte differentiation antigens shared by humans and rhesus monkeys. Anti-human T-lymphocyte serum (anti-HTLA) was used to investigate the relationship between differentiation antigens of human (HTLA) and Macaca lymphocytes. The percentage of lymphocytes sensitive to anti-HTLA is the same in both species, with thymocytes having a higher concentration of HTLA than more mature T-lymphocytes. Macaca bone marrow cells acquire phenotypic properties of T-lymphocytes in vitro in the presence of calf thymic factors. Our data suggest that phylogenetic distinctions between various T-lymphocyte differentiation antigens may be utilized to provide antisera of narrow specificity.


Expression of HLA-B12 on platelets, on lymphocytes and in serum: a quantitative study. Expression of the antigen HLA-B12 on platelets, on lymphocytes and in serum of normal subjects was studied using several quantitative techniques. HLA-B12 was found to be expressed on platelets from each of 25 heterozygous HLA-B12-positive donors in amounts varying about 35-fold from weakest to strongest. Expression of B12 on platelets was enhanced in the presence of the first locus antigen A11, but decreased in the presence of A2, A3, and Aw28. The antigen was expressed in equal strength on lymphocytes sampled from the same donor population. Serum content of B12 assayed in five subjects was found to be roughly proportional to the expression of that antigen on platelets. Expression of B12 on platelets is under genetic control but the precise mechanism of inheritance has not yet been defined. The applicability of these findings to transfusion of single donor platelets to alloimmunized patients is under investigation.


Studies of the mechanism of the periodic acid-Schiff histochemical reaction for glycogen using infrared spectroscopy and model chemical compounds. It has been proposed in the literature that Schiffs reagent reacts with aldehydes to form one of the following types of compounds: alkylsulfonic acids, N-sulfinic acid derivatives, or Schiff bases. Model compounds whose structures are consistent with those proposed in the literature have been synthesized and subjected to infrared analysis. Also, actual products of Schiff reagent reactions with various aldehydes have been isolated and examined using infrared spectroscopy. Comparison of the spectra of the model compounds with those of Schiff-aldehyde reaction products yielded the following conclusions: 1. The reaction of simple organic aldehydes with Schiff's reagent produces an alkylsulfonate-type reaction product. 2. The reaction of periodate-oxidized glycogen with Schiff's reagent probably involves the formation of an alklsulfonate-type compound. 3. The product of the Schiff-aldehyde reaction exists as neither an N-sulfinic acid nor a Schiff base derivative of the fuchsin molecule.


A silver impregnation method for nervous tissue suitable for routine use with mounted sections. A simple, reliable silver impregnation method for nervous tissue is described for tissues fixed in various fixatives including formalin, Bouin, and Susa. Sections are impregnated in a solution containing 1 g Protargol, 2 ml of a 1% Cu(NO3)2 solution, 2 ml of a 1% AgNO3 solution, and 2-4 drops 30% H2O2 in 100 ml distilled water. Sections are impregnated 2-5 days at 37 C and thereafter reduced in a hydroquinone-formalin solution. This is followed by gold toning and subsequent reduction, dehydration and mounting. This method has been found to be very reliable and selective.


Phase-partition fixation and staining of Drosophila eggs. Aqueous solutions of alcohol-acetic acid-formalin or glutaraldehyde-acrolein were shaken with heptane and heptane phase used for fixation. Phase-partition fixation is akin to fixation with vapor. The organic solvent, immiscible with water, penetrates hydrophobic membranes and carries the fixative in contact with water phase of the tissue. Only the fixative enters the tissue, without changing the ionic and water-soluble substance concentrations in the tissue. The quality of this fixation for optical or electron microscopy was as good as that of any conventional fixation method. Staining with basic fuchsin after 2 N HCl hydrolysis gave brilliant staining of nuclei, more intense than that with Feulgen reagent, while cytoplasm remained nearly colorless. Fixing and staining procedures for Drosophila eggs are given.


Triple staining of normal and degenerating nervous tissue. A method of counterstaining sections impregnated according to a previously reported modification of the Glees silver impregnation is described. The basis for this counterstain is the Klüver-Barrera luxol fast blue technique. The results are illustrated and the advantages and disadvantages of the procedure are discussed.


[Chemotherapy in non-Hodgkin's lymphomas (author's transl)]. The type of therapy in non-Hodgkin's lymphomas (NHL) depends on the state of the disease and on histological classification. As in Hodgkin's lymphoma radiotherapy is indicated with localized disease, whereas chemotherapy is given in disseminated cases. In contrast to Hodgkin's lymphoma chemotherapy is indicated earlier (already in most stage II cases), since dissemination may occur outside the typical lymph node areas. Division of NHLs according to higher or lower degree of malignancy is possible histologically on the basis of the Kiel classification as well as of the Rappaport classification. Low-grade malignant lymphomas are treated best with conventional doses of an alkylating agent. An intensive combined chemotherapy is not indicated. Because of the tendency to early and frequent relapses a cyclic maintenance therapy with an alkylating agent is incidated. High-grade malignant lymphomas have to be treated with agressive combination chemotherapy in order to get the patient into complete remission with the possibility for a long relapse-free interval. Up to now there is no indication that any of the published chemotherapy combinations is definitely superior to the others. It is most important to apply the therapy consequently for at least six months. The value of a maintenance therapy is not yet proven.


[Localization diagnostics of malignant tumors using radioactive bleomycin (author's transl)]. Total-body scintigraphy with 111In-labeled bleomycin was performed in 66 patients with malignant solid tumors and systemic diseases of the lymphatic pathways (Hodgkin's lymphoma and non-Hodgkin's lymphoma). In 38 (58%) of 66 cases positive information was correct. In 12 (18%) of 66, scintigraphic information was correctly negative. With 16 out of 66 (24%), false positive or false negative informations were obtained.


Tuberculosis in Tanzania: a follow-up of a national sampling survey of drug resistance and other factors. A total of 1873 patients admitted to a random sampling survey in 15 of the 61 administrative districts in Tanzania in 1969 has been followed up at 1 year or later. The random sample included districts with a long-established tuberculosis service (A districts), those with a service of recent inception (B districts), and those with no specialized tuberculosis service (C districts). The main follow-up concerns 1607 patients with pulmonary tuberculosis of whom 693 had a positive culture at the initial survey, 557 a negative culture and 357 had not produced a specimen. At 1 year or later 12% of the 1607 patients were lost from observation, 10% were alive but with no specimen or no result, 60% were culture-negative, 5% were culture-positive, and 12% were known to be dead. The proportion of patients known to be dead was similar in the 3 types of service, but the proportion lost from observation was highest in the B districts, 24% compared with 7% in the A and 10% in the C districts. Most of the losses occurred early, 73% within the first 3 months. Of the 693 patients with positive culture initially 9% were culture-positive at 1 year, as were 1% of the 557 culture-negative initially. The estimated proportion culture-negative at 1 year for the patients culture-positive initially was highest in the A districts, 78%, and very similar in the B and C districts, 66% and 67%, respectively. The policies of therapy were studied in 1459 patients; 86% were treated in hospital initially for a mean duration of 63 days. The standard regimen of streptomycin, thiacetazone and isoniazid was prescribed in 78% of the patients initially, the proportions being 93% in the A, 48% in the B and 81% in the C districts. The proportion of patients who received or collected supplies of medicament for the full 12 months was only 35%, the proportions being 40% in the A, 20% in the B and 39% in the C districts.


Kodirex for high resolution electron microscopy. Measurement of the important performance parameters shows that Kodak Kodirex film is more suitable than conventional ones for dark-field transmission electron microscopy of molecules at very high mignification. Results are cited for 120 kV; but the relation ship is valid up to 3 MV.


Radioimmunoassays of ethinyl-norgestrienone (R-2323) and medroxyprogesterone acetate (MPA) and their clinical applicability. Radioimmunoassays for 2 synthetic progestins (Ethinyl-norgestrienone, R 2323 and medroxyprogesterone acetate, MPA) are demonstrated. 10 patients aged 31 to 72 years were treated with ethinyl-norgestrienone with different schedules and 3 men suffering from benign prostatic hypertrophy were treated with medroxygesterone acetate. Plasma levels of testosterone, LH, FSH were monitored before, during and after treatment.


[Indication for the surgical treatment of bladder tumors (based on 10-year statistics)]. On the basis of statistics collected over ten years, indications for the surgical treatment of bladder tumors are arrived at. The scopes of the individual surgical approaches, such as transurethral resection and partial and total cystectomy, are discussed. If the preoperative diagnosis indicates a carcinoma that has infiltrated the musculature, it would be disastrous to let the most favorable time for cure slip by while treating with palliative measures a carcinoma that otherwise could have been definitively cured. On no account should a cystectomy represent the last, desperate attempt at therapy when all other measures have failed.


High-resolution metal replication of macromolecules. By adjustment of various parameters affecting crystallite size in thin metal replica films used for contrasting biological macromolecules for electron microscopy, improvements in the level of image information retrieved are demonstrated. Mass thickness, replica metal type, substrate temperature and composition are found to be important determinants of replica quality and conditions are discussed and tabulated for optimizing these. Typical correction curves for replicas are given for platinum and tungsten, from which approximate true dimensions may be calculated from measurements of unknown replicas. Deviation of evaporant sources from ideality are quantitated for both heated filament and electron gun sources. Metal replicas exposed to 80 kV electron beams long enough receive 10(6)e/nm2 show no observable changes other than those attributable to contamination as a function of time after the first few seconds. A variety of artifacts occasionally produced in replicas are discussed in relation to image interpretation.


[Bladder carcinoma: therapeutic concept of the Urology Department of the University Hospital, Mainz]. A therapeutic concept based on tumor staging and grading is presented: T0N0M0 - urine cytology positive - cystoscopy every 3 months. Transitional cell carcinoma (90%): T(iS)N0M0 - carcinoma in situ - cystoscopic biopsy every 3 months. Cystectomy with commencing tumor infiltration. T1N0M0 (80% of all bladder tumors): T1N0M0G0 - TUR; cystoscopy every 3 months. T1N0M0G1-3 - TUR; control-TUR 6 weeks later with systematic biopsy. G3 with tumor recurrence - cystectomy. T2N0/N1M0; G1-2 - TUR; local chemotherapy (adriamycin). G3 - cystectomy; high voltage treatment in inoperable patients. T4NxMx - symptomatic-palliative therapy: TUR, urinary diversion. Squamous cell carcinoma (2-5%): as transitional cell carcinoma; with high voltage therapy adjuvant chemotherapy using bleomycine. Adenocarcinoma (2-3%): as transitional cell carcinoma; cystectomy including part of the anterior abdominal wall and umbilicus. Immunostaging (assessment of the immunocompetence) should be part of the diagnostic procedures and follow-up examination.


[Treatment of cancer of the bladder (author's transl)]. Therapeutic procedures for bladder cancer as practiced at the Department of Urology, University of Vienna Medical School, are presented. Transurethral resection is preferred over in comparison to open surgery. The surgical procedures are always followed by postoperative radiotherapy. The possibility of hemostasis in inoperable carcinomas by bilateral intravasal occlusion of the iliac arteries is emphasized.


[Control of therapy for hyperthyroidism. Comparison of in-vitro and in-vivo methods]. 55 patients with treated hyperthyroidism (thyreostatics) underwent a T3 suppression test and a TRH stimulation test. 60% of the patients had a positive and 40% a negative T3 suppression test. While patients with a positive T3 suppression test always showed a positive TRH test, the behaviour of the TRH test in negative suppression test was different (68.2% positive, 31.8% negative). These discrepant findings are to be explained from the establishment of different phases of regulation. In negative TRH test the concentrations of hormones were significantly higher. Increased basal TSH values are an expression of an overtreatment with thyreostatics. In positive suppression test and TRH test after a therapy lasting at least one year the medication can be stopped. In negative suppression test, however, the therapy should be changed. The TRH test allows only evidence about the quality of the regulation of therapy. Finally it can be established that the two tests on account of their different working points in the regulatory system are necessary and not changeable between each other.


[Arthropathies in intestinal diseases].

[Arthropathies in intestinal diseases]. Together with gastroenterologists rheumatologists describe diagnostically significant disturbances of the supporting apparatus and the locomotor system which frequently occur in ulcerous colitis, enterocolitis regionalis Crohn, Whipple's disease and non-tropical sprue. Important conclusion for the diagnostic approach supplement the description.


[Umbilical cord AFP values in twins (author's transl)]. The concentration of alpha-fetoprotein (AFP) in the umbilical cord in twins was determined via radioimmunological analysis and compared with the values obtained in single births. Since a close correlation exists between AFP content and gestation age, this method can be used to estimate maturity. The mean concentration of AFP was significantly higher in the umbilical blood of fraturnal twins than in that of identical twins. Genetic factors could play a role here.


[Alphafetoprotein in serum of pregnant women. A new assay and practical aspects in prenatal screening for fetal neural tube malformations (author's transl)]. A new radioimmunoassay for alphafetoprotein was tested with 440 sera for it's suitability in the screening of pregnant women for fetal neural tube malformations. The test was reliable and it's execution simple and little time consuming. The test allowed to establish the normal range during pregnancy as well as to detect typical deviations from normal in cases of abortion and anencephaly. Results of mothers with diabetes or elevated transaminase activities were in the normal range. The practicability and acceptable cost of such a test render now possible, and at the same time urgent, a regional screening of women from the 15 th to the 18th week of pregnancy. Only in this way, the detection rate of fetal neural tube malformations can be measured and the usefulness of a general screening of all pregnant women can be evaluated.


[To the determination of chlorinated hydrocarbon pesticide residues with capillary chromatography (author's transl)]. A procedure for determinting chlorinated hydrocarbon pesticide residues by gaschromatography with capillary columns in feeds is described. With the capillary column (OV 61) used a good resolution of all tested 15 pesticides was possible. Recovery and reproducibility were checked, detection limits are given.


[Medical check-up concerning cancer of the prostate. 1st report of experiences, results and documentation of findings]. It is reported on 1,790 medical check-ups concerning cancer of the prostate gland. We found 8 carcinomata = 4.47%. Apart from this 16 adenomata to be operated with adequate disturbances of the micturition were established. The documentation of the findings of medical check-ups by means of a reference on the basis of perforated cards is described and it is referred to the necessity of a systematic documentation of findings.


[Value of cryogenic surgery in the therapeutic plan for prostatic adenoma]. The methodical and instrumental development in urology has led to the fact that nowadays only in 25% of the patients with prostatic adenoma an "open" adenomectoy of the prostate is performed. The cryosurgery is exclusively reserved to high risk patients who at present, however, increasingly also undergo a transurethral resection. The advantages of the transurethral electroresection compared with the cryosurgery of the prostatic adenoma are discussed and confirmed with the help of the own cases. Advanced cerebral sclerosis means contraindication for every operative procedure in adenoma. Nowadays in the high risk patient a transurethral resection should be performed by the best and most experienced urologist.


[Modification of the immune reaction by antigen-immunosuppressive-agent conjugates. I. Tentative hypothesis for the induction of antigen-specific suppression by antigen-immunosuppressive-agent conjugates]. The most important mechanisms for the specific depression of immune reactions--immuno-tolerance, enhancement, transfer of antibodies, drug induced tolerance, immunological suicide, application of antibody-toxin-complexes--are discussed with regard to their possible application in the clinical practice. A tentative hypothesis for induction of antigen specific suppression is proposed, basing on the use of antigen-immunosuppressive agent-conjugates (AIC). Antigen binding lymphocytes are supposed to bind the AIC and to pick them up through endocytosis. After breakdown of the AIC in the lymphoid cells the free immunosuppressive agent can become effective causing damage to the specific cell clones.


[Modification of the immune reaction by antigen-immunosuppressive-agent conjugates. II. Immunogenicity of antigen-immunosuppressive-agent conjugates]. The maintenance of specific immunogenicity of carrier proteins is a necessary condition for the successful use of antigen-immunosuppressive agent-conjugates (AIC) for an antigen specific suppression of the immune response. The experimental results indicate that, in spite of the binding of 6-mercaptopurine (6-MP) and toluyl (T) to bovine gamma globulin (BGG) and human serum albumin (HSA), the carrier specific immunogenicity is not significantly altered. The intradermal application of 6-MP-BGG and T-BGG emulsified with complete Freund's adjuvant in guinea pigs results, in all cases, in a well detectable anti BGG hemagglutination and precipitation titer. This kind of immunization leads also to a formation of anti 6-mercaptopurine and anti new antigen determinants (NAD's) antibodies.


[Modification of the immune reaction by antigen-immunosuppressive agent-conjugates. III. Physiocochemical, antigenic and functional properties of 6-mercaptopurine coupled carrier proteins]. 6-mercaptopurine (MP) conjugates of bovine gamma globulin (BGG) and human serum albumin (HSA) with increasing numbers of MP-groups per protein molecule were prepared and some physicochemical and antigenic properties studied. The electrophoretic mobility increased proportionally to the degree of substitution. In the same manner the formation of high molecular weight aggregates increased. A loss of BGG and HSA specific determinants was observed too. The physicochemical and antigenic properties of these antigen-suppressive agent-conjugates with low and medium coupling degree (to MP26-BGG and MP20-HSA) were similar to the unmodified antigens. The affinity of rabbit anti-DNP-antibodies decreased from K0 = 1,4 - 10(6) M-1 of the unmodified antibodies to 6,5 - 10(5), 3,6 - 10(5) and 3,4 - 10(5) M-1 of the antibodies conjugated with 15, 21 and 29 MP-groups per antibody molecule. The precipitation capacity of the MP-conjugated antibodies decreased to 89, 85 and 61% for the same coupling ratios. These results suggest that the upper limit of the coupling ratio of the antibodies is 20 to 1.


[Modification of the immune reaction by antigen-immunosuppressive-agent conjugates. IV. Studies on the specific suppression of humoral immune response in guinea pigs by antigen-immunosuppressive-agent conjugates]. Bovine gamma globulin (BGG) antigens were modified by the binding of 6-mercaptopurine and toluyl residues, and their influence on the humoral immune response in guinea pigs was investigated. The antigen-immunosuppressive agent-conjugates (AIC) were different, depending on the method used for their preparation and the number of coupled residues per one molecule of BGG. Conjugates denoted as MPI-n-BGG were prepared by special chemical binding of corresponding thioisocyanates. MPII-n-BGG were synthetized by acetylation, and MPIII-n-BGG conjugates, by reductive alkylation. Pretreatment of guinea pigs with MPIII-19-BGG, MPII-16-BGG resulted in a stimulatory effect on the subsequent humoral immune response induced by BGG application. A significant suppressive influence was detectable if the animals had been pretreated with MPII-6-BGG and MPI-26-BGG. MPI-13-BGG and MPI-36-BGG had no effect on the later induced anti-BGG antibody formation. The immune response against a second antigen (human serum albumin) was not influenced by this kind of pretreatment of the animals. Therefore it seems justified to conclude that both stimulatory and suppressive effects seen here were antigen specific and that both the method for chemical modification and the number of coupled 6-MP residues are very important for their effectivity.


[Studies on the significance of antigen in the 6-mercapopurine-influenced humoral antibody response in guinea pigs]. The mechanism of suppression of humoral immune response to dinitrophenylated bovine gamma globulin (DNP23-BGG), human serum albumin (HSA), and trinitrophenylated sheep red blood cells (TNP-SRBC) by 6-mercaptopurine (6-MP) was studied in guinea pigs. Following the intradermal application of the antigens emulsified in complete (CFA) or incomplete Freund's adjuvant (IFA) each test animal was given 6-MP, 10 mg/kg/day for 7 days. This treatment resulted in a significant suppression of the anti BGG and anti SRBC agglutinating and complement binding antibody production. The latter was only significantly suppressed if the TNP-SRBC were applied together with CFA and not if TNP-SRBC were given in IFA. The anti DNP and anti HSA antibody formation was not influenced.


RNA-dependent DNA polymerase activity in intracytoplasmic type-a particles of mouse mammary tumors. Intracytoplasmic type-A particles isolated from mouse mammary tumors and purified by repeated gradient centrifugations exhibit both endogenous and exogenous (template-stimulated) RNA-dependent DNA polymerase activities. The enzyme has a preference for magnesium ions.


[Studies on the immunologic specificity of the macrophage-electrophoresis mobility test in mice with syngeneic and allogeneic transplants of mammary carcinoma]. The study was performed to answer the question as to whether the macrophage electrophoretic mobility test (MEM-test) is suitable to detect allogeneic and syngeneic lymphocyte sensitization in the murine mammary tumour host system. Lymph node cells of both CBAf and XVII/Bln mice injected with allogeneic or syngeneic mammary tumour cells reacted specifically to hypertonic KCl extracts from allogeneic or syngeneic mammary tumours. Lymph node cells of nontreated control mice did not react at all. The MEM results reflect lymphocyte sensitizations which are specific with respect to the claim of transplantation and tumour immunology in the system employed.


[Evaluation of the efficacy of anti-arrhythmic drugs using a computer]. To palliate the deficiencies of the oscilloscopic monitoring of arrhythmias, an automatic system has been developed. It has been used during more than one year time in the Coronary Unit for supervision of patients and for study of the pharmacological effects of antiarrhythmic drugs.


[Choice of surgical procedure in the treatment of stomach cancer]. There are two demands that have to be satisfied in the treatment of gastric carcinoma: 1) eliminate the tumor radically and 2) to maintain the physiological function of the digestive tract. Out of radical methods this is enabled by subtotal and total gastric resections of Billoroth I type. Especially satisfying among the subtotal resections are Billroth I as modified by Shoemaker, Billroth I -- Kirschner (superior and inferior tubular resection) and Völcker, while among the total resections we find Bigham, Longmire and Tomoda I very promising. When dealing with incurable gastric carcinoma palliative methods as gastroenteroanastomosis, recestions and fistulae are used.


Retrograde axonal transport of horseradish peroxidase. Uptake at mouse neuromuscular junctions following systemic injection. Eye muscles were examined histochemically for the presence of horseradish peroxidase (HRP) activity after intravenous injections of large doses of this tracer protein. HRP had penetrated through the blood vessels and diffused into the areas of motor end plates, where it outlined axon terminals at the synaptic clefts. HRP was incorporated into pinocytotic vesicles in the axons from where an intraaxonal transport in the retrograde direction to the nerve cell bodies in the brainstem followed. Accumulation of HRP in perikarya of motor neurons can therefore be the result of a physiological process of pinocytosis at the axon terminals. In this way exogenous macromolecules in the blood can by-pass the blood-brain barrier and reach the lower motor neurons in the CNS.


Repair in arterial tissue. The role of endothelium in the permeability of a healing intimal surface. Vital staining with Evans blue and silver-staining of the aortic intima after a single dilatation trauma. The role of the endothelium as a factor determining the permeability of a healing aortic intima after a severe mechanical lesion in the rabbit was studied by vital staining with Evans blue, by surface light microscopy after silver-staining, fluorescence microscopy and by other methods. Endothelium to develop from pre-existing endothelium in the mouths of intercostal arteries and at the borders of the lesion, remained unstained by vital staining, whereas denuded surfaces as well as pseudo-endothelium, probably derived from smooth muscle cells in the neointima, were intensely coloured. There was a general tendency towards a decreasing permeability of the aortic wall throughout an experimental period of 6 months. The determinant factor in the decreasing permeability was the presence of genuine endothelium, but changes in the endothelium, probably in the form of differentiation of intercellular junctions and changes in the connective tissue in the deeper layers, seemed to be additional factors which influenced the permeability.


Spontaneous mammary carcinoma in C57BL and C3H mice: a histochemical study. The mucosubstances in a series of 12 mammary carcinomas arising in C57BL mice were compared with those in tumours in nine C3H mice and with normal "resting" and lactating glands. A strain difference rather than a difference according to tumour type was found. The epithelial elements in the C57BL tumours contained mainly sialomucin with a little neutral mucosubstance and sulphomucin was present in some. Sulphated mucopolysaccharide was prominent in the stroma of these tumours. The C3H mice tumours contained no sulphomucin, less sialomucin but more neutral mucosubstance. The mucosubstances were studied in serial transplants of four tumours. An acid to neutral mucosubstance change with dedifferentiation of the tumour has been observed and found to be similar to a series of changes previously observed in a group of human gastrointestinal tumours.


Glutaraldehyde fixation for electron microscopy of needle biopsies from human livers. A study including light- and electron microscopy of needle biopsies from normal and fatty human livers fixed by immersion into glutaraldehyde is presented. Four zones which can be detected by light microscopy of toluidine blue stained sections are found: zone 1, the outer one is presumably mechanically damaged. Zone 2 is usually considered to be the most well-preserved region, whereas zones 3 and 4 present increasing swelling of mitochondria, a progressive condensation of microbodies and an increasingly pronounced vesiculation of the smooth endoplasmic reticulum (SER). The alterations observed in zone 3 and 4 are of types which might simulate pathological changes. An increasing irregularity of the outline of the nuclei and a greater accumulation of chromatin along the nuclear membrane is also observed in these two zones. The perimembraneous cytoplasmic ground substance appears to be more dense and the intercellular spaces to be less distinct in the deeper zones in which the path of diffusion of the fixative is long. As regards the penetration into the tissue of glutaraldehyde, normal and steatotic livers were not found to differ. Fixation at 0 degrees C was found to be less effective than fixation at 22 degrees C.


Alcoholic hyalin, microfilaments and microtubules in alcoholic hepatitis. The alcoholic hyalin which is composed of light and dark occasionally swollen and conglomerating filaments, is found to be surrounded by proliferated RER, hypertrophied Golgi apparatus and mitochondria containing enlarged matrical granules. Besides, microfilaments and some microtubules are seen in relation to the hyaline bodies. In biopsies where hyaline bodies are scarce, hepatocytes without alcoholic hyalin present similar changes and the Golgi apparatus is seen to contain very low density lipoprotein-like particles. These ultrastructural changes are suggested to be related to an early stage in the development of alcoholic hyalin. The abundance of microfilaments indicates an increased motility of the hepatocytes, possibly as a part of the regenerative processes. It can hardly be precluded that microfilaments disintegrate and accumulate in the alcoholic hyaline mass.


Peripheral immunofluorescence of hepatocytes. Relation to smooth muscle antibodies and bile canaliculi. By means of indirect immunofluorescence microscopy, a peripheral fluorescence reaction with hepatocytes was found, surrounding the entire cell, with sera from patients with cancer and acute hepatitis and from normal blood donors. It was proved that this reaction was not related to bile canaliculi, contrary to bile canaliculi antibody demonstrated previously from patients with chronic active liver disease. By using fluorescein-conjugated anti-human IgG, the reaction with the periphery of the hepatocytes was produced to a more or less pronounced degree with almost all sera studied. The reaction seems not to be directed against contractile proteins on the liver cell membrane as only a minor part of these sera had also IgG smooth-muscle antibody, and anti actomyosin antibody obtained by affinity chromatografy did not react with the hepatocyte. When applying fluorescein-conjugated anti-human IgM the reaction with the periphery of the hepatocytes was observed in one fourth of the patients with acute hepatitis and only sera which contained also IgM smooth-muscle antibody.


Cell-mediated immune response to guinea pig and bovine basic proteins of myelin in Lewis and PVG rats and their hybrids. The in vitro cell-mediated immune response of rat lymph node cells (LNC) to guinea pig and bovine encephalitogenic protein (EP) has been studied with a LNC transformation test. LNC were obtained from either Lewis, PVG, or F1 (Lewis x PVG) rats 28 days after challenge with guinea pig or bovine EP in Freund's complete adjuvant (FCA) of different mycobacterium content. No differences between the strains or their hybrids in LNC response could be shown if stimulated with sensitizing EP, despite the great differences in capacity for disease development in those animals. Nor did a fivefold lowering of the amount of mycobacterium content in FCA, when guinea pig EP was used at challenge, lessen the in vitro response, although this reduces the disease development. Sensitization with guinea pig EP causes an in vitro cross-reactivity to bovine EP and the reciprocal cross-reactivity probably also exists. In rabbits, this cross-reactivity in both directions was clear-cut. LNC from Lewis rats challenged with guinea pig EP in FAC, with the higher amount of mycobacterium, showed in vitro responses to tested peptides of this EP; the amino acid sequences of these were: 1-42, 43-88, 89-169, and HNB-89-169 (89-169 blocked at the tryptophan-residue by hydroxynitrobenzylation).


[Age-dependence of the cellular immune reaction in childhood. 2. Studies on children with acute lymphatic leukemias]. Results of studies made using the lymphocyte transformation test for leukemic children showed that an evaluation of cellular immunoreactions subsequent to the administration of szecific and nonspecific antigens as well as in the mixed culture is inadequate to recognize the characteristics of age-specific changes. A comparison of the rates of transformation in healthy children with those of leukemic children showed that age-specific characteristics are superseded by disease-specific changes.


[The influence of 6-mercaptopurine on the humoral immune response in guinea pig. 2. The influence on formation of complement binding antibodies (author's transl)]. The effectivity of treatment with 10 mg 6-mercaptopurine/kg/day--beginning on the day of antigen injection and continuing for seven days--on the synthesis of complement binding antibodies is very different. The formation of gamma-2-antibodies against BGG was nearly complete suppressed, an influence on the anti-DNP-gamma-2-antibody formation was not detectable and the antibody formation against HSA was enhanced. The results suggest that the kind of antigen is very important for the immunosuppressive activity of 6-mercaptopurine in guinea pigs.


[Conjugation of bovine gammaglobulin to L-asparaginase (E. coli) (author's transl)]. Bovine gammaglobulin-L-asparaginase conjugates were prepared with an enzymic activity of about 30 IU/mg. Glutardialdehyde was used as bifunctional reagent for the cross-linking in an optimal concentration of 0.16 mg/10 mg protein. The both components were found in the conjugates in a ratio of 1:1.5. The recovery was about 25% of the initial protein amounts. The antigenic specificity of the globulin component decreased up to 60%, the enzymic activity also up to 60% by the conjugation.


[Effect of L-asparaginase on humoral immune response against protein antigens in mice (author's transl)]. The immune response of CBA-mice against alum adsorbed BGG is depressed by daily injections of 20, 40 and 80 I.U. of asparaginase (Crasnitin) from the first day before to the second day after immunisation. After a period of 14 or 21 days the antibody levels slowly rises again. The early IgM-response is not altered. Pretreatment of the animals with BGG-asparaginase conjugate has not any influence on the immune reaction. The conjugate is because its high molecular weight more immunogenic than the BGG and the enzyme allone respectively.


The synthesis of bradykinin-azoprotein and its effectivity. Some authors succeded in binding the bradykinin in form of a haptene by an azo-bridge to the human gammaglobulin as a vehicle. In this way, the bradykinin has been supplemented to a complete antigen. The bradykinin as a strongly vasoactive substance causes--after a local intradermal injection--an intensive circumscribed increase of the permeability of the capillaries. If such animals receive an intravenous or intracardiac injection of a 1% Evans blue solution, there is a strong blue colouring in the place where the bradykinin has been applied. The guinea pigs and the rabbits which had been immunized by bradykinin-azo-protein showed a significant decrease in the local accumulation of the Evans blue-solution,, which must be attributed to the immunologic neutralization of the intradermally injected bradykinin by an anti-bradykinin-antibody. The intradermally injected bradykinin is neutralized by the bradykinin-antibody. The intravenous or intracardiac injection of bradykinin causes a strong bronchoconstriction in the guinea pig. In those animals which have been pretreated with bradykinin-azo-protein, either there was no asthmatic dyspnoea at all or a decrease of the bronchial reaction would be found. This result very probably is also caused by the production and the activity of anti-bradykinin antibodies.


[The influence of 6-mercaptopurine on the humoral immune response in guinea pig. 1. The importance of antigen and adjuvans (author's transl)]. Following an intradermal immunization of guinea pigs with dinitrophenylated bovine gamma globulin (DNP23-BGG), human serum albumin (HSA) and trinitrophenylated sheep red blood cells (TNP-SRBC) emulsified in complete or in incomplete Freund's adjuvans the effectivity of 6-mercaptopurine (6-MP) on the formation of specific antibodies was investigated. By application of 10 mg 6-MP/kf/day--in guinea pigs a significant depression of antibody formation against BGG could be demonstrated if treatment is started on the day of antigen injection and continued for seven days. The production of anti-SRBC agglutinating antibodies was only suppressed if the antigen was injected together with complete Freund's adjuvans. The anti-DNP and anti-HSA antibody formation was not influenced by this kind of immunosuppressive treatment.


[The macrophage disappearance reaction in guinea pigs sensitized with bovine gamma globulin or human scrum albumin (author's transl)]. The macrophage disappearance reaction (MDR) is a suitable test for detection of cell mediated immunity against bovine gamma globulin (BGG) and human serum albumin (HSA) in guinea pigs. The MDR is a technical simple, good manipulable, and quantifiable test. The optimal test conditions for the antigens BGC and HSA are the following: Peritoneal exudat cells (PEC) were stimulated with paraffin oil. On the 5th day after receiving oil the animals were injected with 80 microgram BGG or 30 microgram HSA i.p. 5 hours later the PEC were harvested and counted. With the MDR it is possible to detect differences with respect to degree of cell-mediated immunity. Supernatants of sensitized lymphocytes produces the MDR too.


A delineation of performance criteria for the differentiation of leukocytes. Performance criteria for manual differential leukocyte counting are tentatively set following analysis of the CAP interlaboratory survey programs in hematology. It is evident that intermediate levels of discrimination of five normal leukocyte types (neutrophils, eosinophils, basophils, lymphocytes, and monocytes) can be achieved by almost all laboratories. The most sophisticated level (differentiating band from segmented neutrophils and reactive from normal lymphocytes) is still somewhat controversial. Sampling and other variables when doing 100-cell differential counts at the intermediate level of discrimination are quite predictable, and CAP Survey results closely parallel the predicted results when uniform wedge of centrifuged smears stained in an acceptable manner are used. Automated differential counters should be expected to perform at a level of discrimination at least as good as that of human examiners.


Hepatocellular carcinoma in the Rhodesian African. We have carried out a prospective survey of 28 primary liver carcinomas over one year. Hepatocellular carcinoma is the commonest malignancy seen in Rhodesian blacks, which results in a high index of suspicion and accounts for the 96.4% positive diagnosis before death in this study. The age distribution was evenly spread through adult life with no definite peak incidence. Some were young and without evidence of chronic liver disease, but many had the stigmata of established hepatic disease. This contrasts with the common assertion that in areas of high incidence for primary liver cancer those affected are mainly young and lack signs of chronic liver disease. The commonest presenting symptoms were abdominal pain and swelling and weight loss. Hepatomegaly, often tender and nodular, was present in all but one. The incidence of alpha-feto protein, 46.5%, is low compared with other countries where primary liver cancer is common. Hepatitis B antigen was absent in all 28, suggesting that there is no association between the persistence of the antigen and hepatocellular carcinoma in Rhodesia. Liver function tests, although abnormal, were never diagnostic of primary liver cancer. We have confirmed the association of high alcohol consumption and cirrhosis with hepatocellular carcinoma.


Serum protein polymorphisms in four populations of Afghanistan. Gene frequencies of the serum proteins third component of complement (C3) transferrin (Tf), haptoglobin (Hp), group specific component (Gc), serum cholinesterase (E1), alpha1-antitrypsin (Pi), beta2-glycoprotein I (Bg), and ceruloplasmin (Cp) in the Tajiks, Pushtoons, Hazaras, and Usbeks in Afghanistan were reported. Rare variants were observed in the C3, Tf, and Pi systems.


Modifications of the Goldner and Gomori one-step trichrome stains for plastic-embedded thin sections of bone. Modifications of the trichrome stains by Goldner and Gomori are described. They provide two reliable methods for identifying osteoid seams, bone cells, and other structures in bone embedded in plastic. Trichrome stains for paraffin-embedded soft tissue are not compatible with plastic-embedded bone. These modifications work well with most of the plastic embedding media in routine use.


Hepatitis B antigen: advancement in knowledge quickens. (A review). For many years viral hepatitis and its causative agent have been an enigma to investigators. In the last decade there has been such an increase of information on the subject that it challenges one's ability to keep abreast of the advancement in knowledge. The purpose of this paper is to bring into focus some of the latest developments with regard to viral hepatitis B and its causative agent.


Studies of the antigens involved in an immunologic renal tubular lesion in rabbits. Rabbits injected with nonglomerular components of rabbit kidney incorporated in Freund's complete adjuvant develop a lesion characterized a) extensive interstitial fibrosis, tubular degenerative changes, and sparse focal lymphocytic infiltrates; b) the deposition of IgG and C3 in a granular pattern along the basement membranes of proximal convoluted tubules; and c) functional tubular defects if the lesions are severe. The antibodies were eluted from kidneys with such lesions and labeled with fluorescein isothiocyanate. It was shown that these fluorescein-labeled eluates reacted with the corresponding antigens in the tubular deposits and also with the antigens present in the brush border and/or cytoplasm of the proximal tubules. The antigens are found in proximal tubules of the kidney but not in brain, lung, heart, liver, spleen, bowel, muscles, or urine. They appear to be soluble but may also be present in the plasma membrane.


[Bone biopsy of the iliac crest. Technics of embedding in methacrylate and quantitative readings. Results of 2 years' experience]. The authors report their experience with a method for the study of undecalcified bone embedded in a compound of methacrylate allowing various types of staining procedures. They review the methods of semi-quantitative study of the bone activity and point out their interest in diffuse involvement of the skeleton.


Isolation of a herpesvirus from the cell culture of a malignant melanoma of a ground squirrel (Spermophilus tridecemlineatus). A subcutaneous neoplastic mass in a 13-lined ground squirrel which metastasized to regional lymph nodes and lung was studied. Histopathologically, the tumor architecture and cellular morphology were compatible with that of a malignant amelanotic melanoma. Ultrastructurally, the neoplastic tissue was composed of oval cells, spindle-shaped cells, and spindle-shaped cells with electron-dense cytoplasmic granules. Virus particles were not seen in these cells. Cell cultures from neoplastic tissue grew in complete monolayers and on initial passages contained a few herpesvirus particles. Secondary monolayer cell culture, when exposed to 5-iodo-2'-deoxyuridine or made into several serial subculture passages, caused the appearance of cytopathic effect and the demonstration of many virus particles. The ground squirrel agent, because it contained DNA, was sensitive to chloroform treated and had herpesvirus characteristics on electron microscopy, was considered a herpesvirus. The buoyant density of the virus was 1.298 g/cm3 and the diameter of the enveloped virus particles was 146 nm. This ground squirrel herpesvirus was antigenically distinct from other known herpesviruses.


Results at 5 years of a controlled comparison of a 6-month and a standard 18-month regimen of chemotherapy for pulmonary tuberculosis. The relapse rate up to 5 years after the start of chemotherapy with a 6-month regimen of daily streptomycin, isoniazid, and rifampin has been compared with the rate for a standard 18-month regimen of streptomycin, thiacetazone, and isoniazid for 2 months, followed by thiacetazone and isoniazid for 16 months daily. There were 5 bacteriologic relapses in 145 patients on the 6-month regimen, 1 occurring between 3 and 5 years after admission compared with 3 in 125 patients on the 18-month regimen, 1 occurring between 3 and 5 years.


[Pharmacologic complications of the unequal reactivity of constituent elements of the myocardium to the parasympathetic system]. Pharmacological consequences of the unequal reactivity of the myocardial different parts to parasympathic system. The effects of acetylcholine on the various levels of cardiac automatism are studied on open chest dogs with extracorporeal circulation. This drug depresses the sinus node, and more specially the atrio-ventricular node (AV node), but does not alter the His bundle and the Purkinje fibers activity. The sensitivity of the intra-cardiac conduction to acetylcholine also depends upon the considered level of conduction: measured with a bipolar electrode situated upon the His bundle, this one is lowered electively in the atrio-ventricular part. So, the A, V. node can be considered as the elective place of action of acetylcholine on the double point of view of automatism and conduction. Drugs modifying the vagal tonus see their action altered, either in a plus way (antiarrhythmic agents), either in a minus way (cardiac glycosides, halogenated hydrocarbons).


[Histochemical diagnosis in pediatric rectal suction biopsies (author's transl)]. Between March 1972 and Dec. 1974 532 rectal suction biopsies were obtained from 238 children and histochemical processed. Slight haemorrhage was observed in three. Indications were :1) Group A, 185 suspected aganglionism (AChE. and DPNH) activities were studied). 2) Group B, 7 Neurolipidoses (Red-oil-o) and 3) Group C, Post-operative control in 10 aganglionic patients (AChE, DPNH). Aganglionism was confirmed in 60 cases (25.2%) of Group A [AChE (+), DPNH (-)] although there were five false negatives (8.3%). Lipidosis was confirmed in one case of Group B. Four patients in Group C were AChE and DPNH (+). Results are discussed, and this technique recommended taking in mind its minimal morbidity, simplicity and good results.


[Immunofluorescence study of hyaline membranes and problems involved (author's transl)]. Lung tissues from died premature babies were studied by means of the fluorescein technic, using anti-IgG, anti-IgM, anti-fr. C-3 of the complement and anti-fibrinogen. It is shown that fibrinogen, IgG and C-3 fr. of the complement are present in the hyaline membrane. In view of these findings, and upon the available data on the subject and current research, the role of the complement in the production of hyaline membrane is discussed.


Anaplasia rediviva.

Anaplasia rediviva. Anaplasia refers to the apparent retrogression toward an embryonic state that occurs in some human and animal tumors, the consequence, presumably, of a derepression of genes that would normally have been switched off in the course of development. The fetal substances newly formed in tumors are identified immunologically and are therefore referred to as "antigens." Some such antigens, for example, carcinoembryonic antigen and alpha-fetoprotein, and blood-group precursor substances have great promise diagnostically but play no known part in resistance to malignant growth. The reawakened fetal antigens that are of special interest and importance are those that are capable of arousing cell-mediated immunity and thus may contribute to antitumor immunity. Modern research, therefore, rehabilitates an etiologc notion earlier thought to have been discredited.


[The consumer in the face of change].

[The consumer in the face of change]. The study on consumer behaviour done by the nutrition department of INSERM shows that there are opposing attitudes towards Change. Among the factors unfavorable to change are: reluctance to changing old habits, nostalgia for the past, suspicion towards industrially prepared foods, fear of food pollution, love for the art of good cooking. Among the factors favoring change are: the higher standard of living, the desire to save time, the change in the way of thinking about foods, the evolution of cooking practices, as well as a need for variety and quality. Man looks for change more for his own pleasure than by necessity. They can be positive pleasure such as: -- Sensory values (taste, zest, variety). -- Coenesthesic values (feeling of well-being, euphory). -- Emotional values (means of communication a gift). -- Symbolic values (prestige, personnalization, creativity, distraction). However there are also negative pleasures. That is all factors which decrease the effort or the work load such as: -- Health values (harmlessness, safety, maintaining attention). Practical values related to convenience (time saving, ease of storage). The favorizing these positive or negative pleasures will lead to change. The consumer will accept things which give a maximum of pleasure for the least effort. He will also accept something which by its taste, prestige, convenience and safety contributes to a better life.


Novel actinomycins formed by biosynthetic incorporation of cis- and trans-4-methylproline. Streptomyces parvulus (Streptomyces parvullus) normally produces actinomycin D; in the presence of cis-4-methylproline, this species synthesizes two additional actinomycins, designated K(1c) and K(2c), in which one and two proline sites, respectively, are occupied by cis-4-methylproline. Analogously, actinomycins K(1t) and K(2t) are formed in the presence of trans-4-methylproline. Both mixtures were separated chromatographically, and the four novel actinomycins were obtained in crystalline form. Their biological activities were compared with that of actinomycin D in respect to inhibition of ribonucleic acid, deoxyribonucleic acid, and protein synthesis and antimicrobial potency. In all cases examined, the order of activity D > K(1t) > K(1c) > K(2t) > K(2c) was observed, and the same sequence prevailed in a spectroscopic measure of their binding to deoxyribonucleic acid. In addition, proton nuclear magnetic resonance studies revealed that the replacement of proline by cis-4-methylproline alters the conformation of the antibiotic molecule.


Identification of intermediates formed during the degradation of hexachlorocyclohexanes by Clostridium sphenoides. Washed cell suspensions of Clostridium sphenoides degraded the alpha-isomer of 1,2,3,4,5,6-hexachlorocyclohexane via delta-3,4,5,6-tetrachloro-1-cyclohexene and the gamma-isomer via gamma-3,4,5,6-tetrachloro-1-cyclohexene. Both intermediates were further metabolized to unknown substances. The tetrachlorocyclohexene intermediates were identified by gas chromatography and mass spectrometry.


Congenital sensory neuropathy: report of an atypical case. An otherwise normal 11-year-old boy had reately reduced acral pain and temperature sensation with associated trophic damage. The disorder was present at birth, and there was no family history of similar problems. The patient also exhibited complete anhidrosis. The case may an "overlap" between what has been termed as hereditary sensory neuropathy (HSN) type 2 and type 4.


Demonstration of bilateral claustro-cortical connections in the cat with the method of retrograde axonal transport of horseradish peroxidase. The claustrum of the cat was studied to determine the extent of its projection to the cerebral cortex. Neurons in the bilateral dorsal claustrum are labeled by retrograde axonal transport of horseradish peroxidase (HRP) from injection sites of different neocortical areas with preponderance to the ipsilateral side. It is revealed that the dorsal claustrum projects to almost entire regions of the bilateral neocortex and that distribution of labeled neurons in the bilateral claustrum has topographical correspondence, rostro-caudal as well as dorso-ventral, with regard to the sites of injections in the neocortex. No neurons in the ventral claustrum are labeled.


Modified tannin-osmium conductive staining method for non-coated scanning electron microscope specimens. Its application to microdissection scanning electron microscopy of the spleen. Supplement of guanidine hydrochloride to the original tannic acid solution of MURAKAMI (1973) eliminated, without debilitating the conductivity effect, the vexatious treatment with different amino-acids and sucrose in the previous method (MURAKAMI, 1974). The procedures and advantages of this simplified modification are described using human spleen samples; attempt was made to elucidate the structure of arterial terminals by dissecting the specimens under the scanning electron microscope.


Shoulder-hand syndrome in a hemiplegic population: a 5-year retrospective study. From 1969 through 1973, 68 (12.5%) of 540 rehabilitation inpatients  with hemiplegia were diagnosed as having shoulder-hand syndrome. Care was used to distinguish these patients from those with other shoulder pathologic conditions and pain syndromes. Patients were evaluated with respect to side of hemiplegia, dates of onset of hemiplegia and of pain, age, sex, handedness, sensory losses, associated medical diseases and treatment response. All patients became pain-free within three weeks with a therapeutic regimen of low doses of steroids orally, passive range of motion to pain tolerance, use of a hemiplegia sling and the application of physical modalities for symptomatic relief. Losses of range of motion in the affected extremity responded less well to treatment. No complications or side effects attributable to steroids were observed. The full syndrome recurred in six patients, all of whom responded to a second course of treatment.


[A protective hearing aid (author's transl)]. A device consisting of a hearing protector headset and a modified hearing aid is used as a protective hearing aid for normal hearing subjects. Understanding of sufficiently redundant speech is remarkably improved, especially for noise level of about 20 dB above speech level. Tests have been made with the "Freiburger Zahlentest" and amplified "party noise" up to noise levels of 105 dBSPL. As an example: the above mentioned device enables the wearer to understand speech of 85 dB with 105 dB loud speech similar background noise while still the ears are charged with no more than 90 dB at all. Application for hyperacusis and recruitment patients is discussed.


Transperineal intraprostatic injection treatment of benign prostatic enlargement. Thirty-nine patients with benign prostatic enlargement were treated by transperineal intrasprostatic injections composed of phenol 2%, glacial acetic acid 2%, and glycerine 4%, in distilled water. A success rate of 56.41% was achieved, which is much less than that previously reported. Complications were negligible. Follow-up for two years was satisfactory. The patients in whom injection therapy failed could still be treated by any standard operative treatment. Why this mode of treatment was successful in some cases and not in others is not clear.


Chromosomal characteristics of six cultured lymphoblastoid cell lines originating from Marek's disease lymphomas. Cytogenetic observations were made on 6 cell lines (MOB-1, MOB-2, MOB-3, MSB-1, HPRS Line 1, HPRS Line2) originating from Marek's disease lymphomas and 2 clones (1104-B, 1104-X-5) of a cell line established from an avian lymphoid leukosis tumor. The modal chromosome number was within the diploid range in all the lines except HPRS Line 1 and HPRS Line 2, both of which had a mode at about 60. Karyotypes were grossly abnormal in 4 cell lines: trisomy for No. 1 in MOB-2; the heteromorphic No. 1 pair in MSB-1, and marker chromosomes derived from rearrangements involving No. 3 or No. 5 and unidentified elements in HPRS Lines 1 and 2. The MOB-1 line which had been characterized by cells with an apparently normal karyotype was completely taken over by cells with a heteromorphic No. 1 pair morphologically similar to the one found in MSB-1 by the 95th day of continuous growth in vitro. BUdR-acridine orange differential staining technique revealed, however, different banding patterns in these abnormal chromosomes.


Studies on the mode of oestrogenic inhibition of hepatic synthesis of alpha2u-globulin and its corresponding messenger ribonucleic acid in rat liver. 1. The possible mechanism of the oestrogenic inhibition of the androgen-dependent synthesis of alpha2u-globulin in rat liver was explored by a correlative study of the amounts of alpha2u-globulin, its corresponding mRNA and circulating testosterone in oestrogen-treated male rats. 2. Daily treatments of mature male rats with oestradiol-17beta (10 microgram/100g body wt.) decreased and ultimately stopped the hepatic synthesis of alpha2u-globulin as determined by both hepatic and urinary concentrations of the protein. The oestrogen-mediated decrease in the hepatic synthesis of alpha2u-globulin was correlated with a decrease in the mRNA for this protein. 3. Withdrawal of oestrogen resulted in the recovery of alpha2u-globulin synthesis and an increase in mRNA for alpha2u-globulin. 4. At higher doses of oestradiol-17beta (50 microgram/100g body wt.), synthesis of alpha2u-globulin was totally suppressed. In addition, this treatment resulted in an extended period of androgen-insensitivity during which treatment with androgens induced synthesis of neither alpha2u-globulin nor its corresponding mtrna. 5. it is concluded that the oestrogenic inhibition of alpha2u-globulin synthesis is mediated by an oestrogen-dependent decrease in the hepatic content of translatable mRNA for alpha2u-globulin.


Serological analysis of cellular and viral DNA polymerases by an antiserum to DNA polymerase gamma of human lymphoblasts. An antiserum has been prepared against a highly purified DNA polymerase gamma from NC37 cells, a normal human lymphoblast cell line. The antiserum does not possess enzyme neutralizing activity, but does bind specifically to DNA polymerase gamma. When tested in a double antibody immunoprecipitation assay, the antibody does not cross-react with DNA polymerases alpha or beta, purified from NC37 cells, or with reverse transcriptases of avian, murine, or primate RNA tumor viruses. Antisera prepared against purified reverse transcriptases similarly do not recognize DNA polymerase gamma, either in an enzyme neutralization assay or in the more sensitive double antibody immunoprecipitation assay. The availability of an antiserum to DNA polymerase gamma will allow the further characterization of enzyme activities isolated from cellular material and suspected of being related to viral reverse ttranscriptases. In those cases where such activities do not immunologically resemble known viral DNA polymerases, the anti-DNA polymerase gamma will help determine the viral or cellular nature of the unknown activity.


Flash-induced 515 nm absorbance change in chloroplasts with various granum contents. The 515 nm absorbance change was studied in mesophyll and bundle sheath chloroplasts of maize, which contain different amounts of grana. The amplitude of the 515 nm signal (induced by 3 micro seconds flashes repeated at 4 s intervals) has shown a correlation with the granum content of the samples. However, upon addition of N-methylphenazonium methosulphate the 515 nm signal became independent of the amount of grana: in agranal thylakoids a large pool of silent Photosystem I was activated and, as a result, the amplitude of the 515 nm signal of agranal chloroplasts increased to the level exhibited by granal chloroplasts. These data show that the 515 nm absorbance change is not limited to small closed vesicles like grana, but in the presence of suitable electron donors single lamellae of bundle sheath chloroplasts can also be active.


Localization of the basic protein and lipophilin in the myelin membrane with a non-penetrating reagent. The localization of proteins in myelin was studied by the use of a non-penetrating reagent. Tritiated 4,4'-diisothiocyano-2,2'-ditritiostilbene disulfonic acid was used to label the isolated myelin membrane. The membrane was labelled, the basic protein and the hydrophobic protein, lipophilin, were isolated. After 10 min of exposure to the reagent, the specific activity of lipophilin was found to be 10 times greater than that of the basic protein. Water shock did not alter the specific activities. However, sonication increased the specific activity of lipophilin but not that of basic protein. When the isolated proteins were labelled with 3H-labelled 4,4'-diisothiocyano-2,2'-ditritiostilbene disulfonic acid, the specific activity of the basic protein was 10 times that of lipophilin. We concluded that the low specific activity of basic protein isolated from the labelled membrane was due to the inaccessible position of this protein in the membrane bilayer.


Enhancement of opiate binding by various molecular forms of phosphatidylserine and inhibition by other unsaturated lipids. A study was undertaken on the possible involvement of phospholipids on stereospecific opiate binding to a rat brain membrane fraction comprised mainly of synaptic membranes. The addition of acidic phospholipids such as phosphatidylserine, phosphoinositides, and phosphatidic acid significantly enhanced opiate binding. With the exception of phosphatidylserine, when the acidic phospholipids contained a polyunsaturated acyl group, they were actually inhibitory, along with neutral phospholipids derived from brain. Both the C18:0, C18:1 form (derived from myelin) and the C18:0, C22:6 form of phosphatidylserine (derived from synaptic membranes) produced as much as a 45% enhancement in opiate binding. Unsaturated fatty acids were highly inhibitory, the degree of inhibition being related to the degree of unsaturation. Both phospholipase A and C were inhibitory; and the inhibitory effect of A could not be prevented by albumin or overcome with the addition of phosphatidylserine. With the use of the cross-linking agent, dinitrodifluorobenzene, it could be demonstrated that the phosphatidylserine of synaptic membranes appeared to be preferentially associated with membrane protein. The enhancement of opiate binding by phosphatidylserine diminished with increasing degree of cross-linking.


The effect of phenethyl alcohol on in vitro DNA synthesis in Escherichia coli. The effect of phenethyl alcohol on DNA synthesis was examined using several in vitro systems of Escherichia coli H560; i.e., ether-treated cells, membrane fractions and folded chromosomes fortified with DNA polymerase. In all systems, the incorporation of deoxyribonucleotides was much reduced for the phenethyl alcohol-treated cells compared with the non-treated cells. The total activity of DNA polymerases in polA1 cells (mostly DNA polymerase II) was not impaired for the phenethyl alcohol-treated cells and the reduction of the rate of DNA synthesis in vitro was ascribed to the reduction of the chromosomal template activity which was related to trypsin sensitive protein components. The analysis of chromosomes from the phenethyl alcohol-treated cells revealed the remarkable reduction of a protein component of molecular weight approx. 58 000 in contrast with a protein component of molecular weight approx. 30 000.


The peptide antibiotic gramicidin D. A specific reactivator of tyrocidine-inhibited transcription. The sporulating bacterial strain Bacillus brevis (ATCC 8185) produces two types of peptide antibiotics, the linear gramicidins and the cyclic tyrocidines. Their effects on transcription in vitro were studied: 1. RNA synthesis catalized by RNA polymerase and DNA as template, both from B. brevis cells, is inhibited by the addition of gramicidin maximally to 50% as compared to controls without gramicidin. This inhibition is dependent on the concentration of gramicidin as well as on that of RNA polymerase. 2. Transcription of DNA is also inhibited by tyrocidine. This inhibition is partially compensated by the addition of gramicidin. Here, the action of gramicidin is again dependent on its concentration and on that of RNA polymerase. 3. This counteraction of gramicidin occurs only with RNA polymerase from B. brevis. Enzyme preparations of other origin are additively inhibited by gramicidin when previously inhibited by tyrocidine. The specific action pattern of gramicidin and tyrocidine in connection with the B. brevis RNA polymerase supports the notion that the two peptide antibiotics may be involved in a gene regulatory mechanism during sporogenesis.


Characterization of a chemotactic and cytotoxic proteinase from human skin. A proteinase (EC 3.4.-.-) active at physiological pH has been isolated from human skin utilizing gel filtration and affinity chromatography techniques. The proteinase has a molecular weight of approx. 28 000 and it is inhibited by alpha 2-macroglobulin, alpha 1-antitrypsin, C-1 inactivatory, soybean trypsin inhibitor and diisopropyl fluorophosphate. 2njection of 1 ng of purified proteinase into rabbit skin induces polymorphonuclear leukocyte infiltration of the cutis. Inhibition of enzyme activity with diisopropyl fluorophosphate inhibits the chemotactic effect. Addition of 0.2 microgram/ml of purified proteinase to fibroblast cultures kills the cells within minutes. This proteinase may play a significant role in modulating the inflammatory response after cellular injury.


Activatable cholesterol esterase and triacylglycerol lipase activities of rat adrenal and their relationship. Activatable cholesterol esterase and triacylglycerol lipase of rat adrenal were 58-69% recovered in the 100 000 X g supernatant fraction. Activatable triacylglycerol lipase activity was differentiated from the activity of acid lipase and lipoprotein lipase also found in this fraction. Cholesterol esterase was activated 39.7 +/- 13.6% (S.D.) and triacylglycerol lipase 11.9 +/- 2.9% in a reaction dependent on ATP, cyclic AMP, and protein kinase. The two activities were shown by differential inhibition by an organophosphate, and by partial separation on salting out, to be largely due to separate enzymes. The two enzymes bound tightly to substrate emulsions with quantitatively similar distribution between competing emulsions, suggesting concerted binding. Coinciding gel filtration patterns reinforced, The hypothesis of a lipase complex. Cholesterol esterase comprised a major component of higher apparent Km for substrate and molecular weight 3-10(5)-6-10(5) by gel filtration and a minor component of lower apparent Km and heterogeneous molecular weight above 1 million, which was found mostly in complex and lipid.


Evidence suggesting the presence of common antigenic determinant between dynein and tubulin. The present experiments showed that the guinea pig antiserum prepared against the main polypeptides of 14 S dynein from Tetrahymena cilia reacted with sea urchin sperm flagellar dynein and with bovine brain high molecular weight protein to give rise to a precipitin line confluent with that formed between the antiserum and Tetrahymena dynein. Furthermore, it was found that this antiserum also reacted with tubulins from Tetrahymena cilia, sea urchin sperm flagella and bovine brain to give rise to the confluent precipitin line. Among muscle proteins, only actin preparation from rabbit skeletal muscle reacted with the anti-Tetrahymena dynein serum, whereas neither rabbit skeletal muscle myosin, chicken skeletal muscle tropomyosin nor chicken skeletal muscle troponin reacted with the antiserum. These results suggest that dynein and tubulin and probably actin share an antigenic determinant regardless of different protein species and of different animal species. The common antigenic determinant was detected only when the proteins denatured with urea/sodium dodecyl sulfate/beta-mercaptoethanol/N-ethylmaleimide were used, but it was not detected at all when the native proteins were used. This implies that a certain common antigenic determinant which is involved in the precipitin line formation exists in the primary structures of dyneins and tubulins and probably actin, and is hidden inside the tertiary structures of the native protein molecules.


Immunochemical purification and characterization of ovine alpha-fetoprotein. Ovine alpha-fetoprotein was successfully isolated from fetal sheep serum by using rabbit anti-ovine alpha-fetoprotein linked to an agarose immunoadsorbent column. Antibody used in this affinity chromatography column was produced by immunizing a rabbit with highly purified alpha-fetoprotein-antibody complex to yield a monospecific antiserum to ovine alpha-fetoprotein. Following affinity chromatography, alpha-fetoprotein was further purified by preparative polyacrylamide disc gel electrophoresis ultimately yielding a 105-fold purification. The purified alpha-fetoprotein was homogeneous on analytical polyacrylamide disc gel electrophoresis. Ovine alpha-fetoprotein was found to be immunochemically related to human alpha-fetoprotein and to exhibit a molecular weight and amino acid composition similar to other mammalian alpha-fetoproteins.


Subunit structure of human alpha 2-macroglobulin (alpha 2-MG) with respect to its interaction with trypsin. SDS-polyacrylamide gel electrophoresis of a recently prepared alpha 2-macroglobulin solution showed only the polypeptide chains of 190,000 molecular weight. Reduction-alkylation of this preparation followed by gel-filtration on a Sephadex G-200 column in 5.2 M guanidine hydrochloride was unable to separate a fraction of 83,000 molecular weight as previously described. Nevertheless, after incubation of a mixture alpha 2-macroglobulin-trypsin during 45 minutes at 37 degrees C, approximately 60 per cent of the preparation were converted in a component with 83,000 molecular weight as detected in SDS polyacrylamide gel. That component was isolated on Sephadex G-200 in guanidine hydrochloride and corresponds to the subunit, fraction II. According to the results of the present work together with those of previous studies, it can be assumed that alpha 2-MG is a 780,000 molecular weight protein (19S) formed of two half-molecules of equal weight (11-12S). The half-molecule contains two polypeptide chains of 180,000-190,000 molecular weight, each of them having, in its middle, a specific region particularly susceptible to attack by proteases.


Objective identification of cell cycle phases and subphases by automated image analysis. Frequency distributions of integrated optical density, perimeter, projection, area, form factor, average optical density, and mean dispersion path of nuclear images of Feulgen-stained HeLa S3 cells were obtained by automated image analysis at the base threshold of 0.04 OD. The mean values and standard deviations of these geometric parameters were then computed versus increasing values of threshold (0.08--0.32 OD). There is clear evidence of differential chromatin dispersion and convolution during the cycle of synchronized HeLa S3 cells at different times after selective mitotic detachment. The combination of average OD, form factor, and mean dispersion path at base threshold with the threshold dependence of nuclear morphometric parameters permits objective identification of cell cycle phases and their subphases, by characterizing variations in chromatin geometry within and between phases, regardless of whether DNA content remains constant (early G1, middle G1, late G1), varies only slightly (late G1-early S or late S-G2 transitions), or varies significantly (early S-middle S).


[Synthesis of alpha-fetoprotein and albumin human embryo hepatocytes]. Population aspects of specific secreted proteins (alpha-fetoprotein and serum albumin) were analyzed in the cultured human embryo hepatocytes (6 to 12 weeks' gestation). A method based on local hemolysis in gel of sheep erythrocytes conjugated with antibodies specific of proteins in question. The great majority of individual hepatocytes synthesized both proteins.