Effect of the culture medium and incubation time on auxins production by bacteria isolated from the roots of pine seedlings (Pinus silvestris l.). Studies were performed on the effect of culture medium and incubation time on the production of auxins by bacteria. The bacteria studied produced more auxins in the mineral medium containing glucose and tryptophan than in that enriched with casamino acids and yeast extract. The amount of auxins elaborated depended both upon the strain and the age of the culture. Some strains produced the largest amounts of these substances after 7 days of incubation while others required a longer period. Most of the substances showing auxin activity were located on the chromatograms at Rf 0.3-0.4 and 0.8-1.0.


Adenylate energy charge and the steroid 11 alfa-hydroxylase activity in Monosporium olivaceum during growth and starvation. During growth of Monosporium olivaceum its energy charge, E.C., (i.e. the adenylates ratio ATP + 0.5 ADP/ATP + ADP + AMP) increased from an initial value of 0.59 up to 0.85 after 25 hr of growth and then decreased to 0.51. The increase of energy charge was followed by the decrease of the activity of the 11 alpha-hydroxylase of cortexolone. This occured very clearly in the starved mycelium. Highest hydroxylation activity was observed when the lowest E.C. level (0.39-0.33) was reached.


Changes in the cellular content of the pool constituents of Monosporium olivaceum -- a steroid hydroxylating mould. Changes of the metabolic pool constitutents of Monosporium olivaceum -- a mould capable of steroid hydroxylation were examined. The experiments were carried during growth and starvation of the microorganism. The highest activity of the 11alpha-hydroxylase was observed in the mycelium which contained the lowest level of free amino acids, glucose, and mannitol. It is suggested that the inhibition of biosynthetic processes and the decrease of the respiration rate, the activity of the NAD(P)H regenerating systems maintained, provide the optimal physiological conditions for the activity of the steroid hydroxylases.


Physiology and characterization of a fungal milk-clotting enzyme from Aspergillus flavus. Aspergillus flavus produced extracellularly an active rennin-like enzyme when grown aerobically in whey media. The enzyme was detected at early stages of growth reaching a maximum after three to four days at 25 degrees. The activity was destroyed by heating to temperatures higher than 50 degrees, whereas the presence of skim milk during heating preserved the enzyme activity, at least, up to 70 degrees. Calcium chloride significantly stimulated the milk-clotting activity up to 1% final concentration. The clotting time was inversely proportional to protein concentration in the range 0.2-0.6 mg/ml and the enzyme exhibited marked stability when stored at 37 degrees at pH 6.


Preliminary studies on the kinetics of utilization of acetate by bacterioplankton in Mikołajskie Lake. The kinetics of the utilization of acetate by bacterioplankton in natural waters was measured according to a modification of the method of Wright and Hobbie (1965, 1966) which enabled investigations in natural conditions in the open. A number of measurements made with this method allowed the determination of the natural concentration of the substrate in lake water and potential maximal rate and time of utilization of the substrate by bacterioplankton. Measurements were made in different zones and at various depths in Mikołajskie Lake in July, August and Septemper of 1974.


Studies on the biodegradation of nonionic surfactants applied in the polyestre fibre industry. III. Effect of adaptation on the respiratory activity of organisms and on the biocenosis of inoculated sludge. Manometric studies were conducted in 3 series. In the first one the sludge used for inoculation was non adapted to water purification containing surfactants. In the second and third series it was adapted to the sludge load 0.11 and 0.28 mg surfactant/mg d.w./day respectively. Increase in oxygen uptake was proportional to the degree of the adaptation of microorganisms only within limited range of surfactant concentration not exceeding 500 mg/l for Cirrasol FP and 1000 mg/l for Cirrasol SF and Cirrasol TCS. The toxic action of surfactants against Ciliata was noted at concentration above 100 mg/l and for Mastigota and Sarcodina above 500 mg/l, even by using adapted sludge for inoculation.


Control of proliferative growth in androgen responsive organs and neoplasms. Growth of a normal androgen-responsive organ appears to be ordered by the function of three constraint mechanisms which are sensitive to the intranuclear concentration of androgens. For the complete expression of these constraint mechanisms, several properties underlying hormonal responsiveness must be manifest by the cell, including the presence of cytoplasmic receptor, the ability to transfer androgens into the nucleus, the competence to form nuclear receptor, and the fidelity of the interaction between androgens and chromatin. Cytoplasmic receptor alone is not an exclusive indication of hormonal dependence in vivo, but its presence is associated with enhanced ability of the cell to incoropate androgens into the nucleus. Androgens are required for the initiation of DNA synthesis and cell proliferation, and nuclear receptor may not be required for these responses. On the other hand, it is possible that the function of the latter molecule is concerned with negative feedback or cellular autolysis.


Antiandrogens.

Antiandrogens. Many antiandrogens, mainly steroidal and some nonsteroidal agents, have been synthesized and tested in several available biological assays. Unfortunately, many of these compounds have other biological activities which make it difficult to ascertain the precise mechanism of antiandrogenic action. The blocking of androgen action can be accomplished by a number of ways: (1) the inhibition of gonadotropin release and/or synthesis, (2) the interference with testosterone and/or dihydrotestosterone biosynthesis, (3) the blocking of protein synthesis, and (4) the competition with androgens at receptor sites. Although the major reason for the development of antiandrogens is to utilize them in certain clinical situations, some have become important tools in studying androgen action, particularly on the molecular level. The clinical effectiveness of some antiandrogens in prostatic hyperplasias, hirsutism, and acne represents an important advance in therapeutics, but the search for more potent antiandrogens with minimal side effects should continue.


A competitive-inhibition radioimmunoassay for influenza virus envelope antigens. A double-antibody competitive-inhibition radioimmunoassay for influenza virus envelope antigens is described. A viral antigen preparation from influenza A virus recombinant MRC11 [antigenically identical to A/Port Chalmers/1/73 (H3N2)] consisting of haemagglutinin and neuraminidase was labelled with radioiodine. Rabbit antisera were allowed to react with the labelled antigen and the resultant antigen-antibody complexes were precipitated with the appropriate antiglobulin. The competitive-inhibition radioimmunoassay was very sensitive in elucidation of differences even among closely related influenza virus strains. Attempts have been made to eliminate neuraminidase from radioimmunoprecipitation in order to obtain a competitive-inhibition radioimmunoassay system for haemagglutinin alone.


Persistent SV5 virus infection in continuous cell cultures. A continuous line of guinea pig kidney cells (CGPK/H) and a continuous line of mouse fibroblasts (L/H) spontaneously infected with parainfluenza virus SV5 were found. These cultures showed no enhanced cell degeneration or symplast formation, nor was haemagglutinin accumulation or infectious virus demonstrated in them. However, regular reproduction of ribonucleoprotein (RNP) characteristic of parainfluenza viruses, morphologically complete virions and antigens producing antibody to SV5 virus were found in the cells. Focal haemadsorption neutralized by antiserum to SV5 virus was also demonstrated. The infection persisted in the cell populations for over 2 years (the observation period) under standard conditions of cell dispersion and subcultivation.


Two variants of concealed trigeminy.

Two variants of concealed trigeminy. Long rhythm strips were analyzed from three patients with frequent unifocal ventricular extrasystoles. The predominant rhythm in each patient was concealed trigeminy; i.w., the number of conducted sinus beats, S, between extrasystoles satisfied the equation S = 3n --1, where n is any positive integer. In one of these patients, about one-fourth of the values of S did not satisfy this equation; however, all such exceptional values of S satisfied the equation S = 3n. In the other two patients, very few of the exceptional values of S fit the equation S = 3n, but they did conform to the equation S = 3n --2. It is proposed that in all forms of concealed trigeminy, there is a characteristic 3:1 block in the re-entry loop that is responsible for the extrasystoles. Furthermore, it is postulated that immediately after each manifest extrasystole, the 3:1 block is converted transiently to a 4:1 or 2:1 block in the "3n" and "3n--2" variants, respectively. After the first penetration of the block site after a manifest extrasystole, if the re-entrant impulse is concealed, the 3:1 ratio is resumed in both variants until the next manifest extrasystole appears.


Intermittent parasystole with concealed extrasystolic bigeminy during myocardial infarction. A 72-year-old man demonstrated persistent intermittent ventricular parasystole for 6 days during an acute inferior wall myocardial infarction. Entrance block failure occurred at a critical interval of 1.24 sec. after parasystolic beats with resetting of the parasystolic cycle length. The first ectopic beat in each parasystolic series showed coupling to its second preceding conducted beat, and successive cycle lengths in each series usually showed gradual shortening. Concealed extrasystolic bigeminy was also demonstrated between parasystolic series with the number of intervening conducted beats conforming to the formula 2n + 2. After interpolated parasystolic beats, this formula became 2n + 3.


identification of Cryptococcus neoformans in cytologic preparations of cerebrospinal fluid. Review of routine Papanicolaou-stained cerebrospinal fluid preparations from 13 patients who had meningeal cryptococcosis documented by other methods demonstrated the yeast in 11 cases. Special stains greatly facilitated the detection of the organisms in two samples and discriminated them from artifacts. An increased number of cells was present in nine cases. Correlation with the clinical data revealed that every patient but one had a malignant lymphoma, most commonly Hodgkin's disease. The exception was a patient who had disseminated carcinoma of the breast treated with adrenal corticosteroids. The clinical history and the cellularity of the smear should alert one to the possibility of cryptococcosis.


Orosomucoid contents of pleural and peritoneal effusion of various etiologies. Orosomucoid and the contents of seven other proteins were determined by radial immunodiffusion in sera and in pleural or peritoneal fluids of 66 patients. No significant difference between concentration of individual proteins in effusions of neoplastic origin and those of nonneoplastic inflammatory origin was found. In noninflammatory fluids, the protein concentrations were lower, but their distribution was similar to distributions in the other fluids tested. These findings support the view that the protein content of effusions reflects changes in vascular permeability.


Serum alpha-fetoprotein in adults, in women during pregnancy, in children at birth, and during the first week of life: a sex difference. In order to investigate whether a sex difference exists in alpha-fetoprotein serum concentration, blood samples were collected from 133 adult men, 52 adult women, 239 pregnant women at different ages of gestation, 16 girls and 18 boys at birth, and seven girls and seven boys during the first week of life. In the case of 15 boys and 10 girls, blood samples were also collected from their mothers at the time of delivery. Serum AFP was measured by a radioimmunoassay method. In all neonates the mean value was 625 times higher than in all mothers at the time of delivery and 13,000 times higher than in adults. No difference was found between mothers of girls and those of boys both during pregnancy and at the time of delivery. However, at birth, the fetal AFP serum concentration was 1.8 (p less than 0.001) times higher in boys than in girls. This sex difference was maintained during the first week of life. The half-life of serum AFP was 5 to 6 days in the neonate. The origin and the possible physiologic significance of such sex difference in AFP deserve further investigations.


A clinical and laboratory evaluation of immune serum globulin from donors with a history of hepatitis: attempted prevention of post-transfusion hepatitis. A controlled trial of passive immunization for prevention of post-transfusion viral hepatitis was carried out in order to determine whether effective levels of antibody were present in the "convalescent" immune serum globulin used in the study. This globulin was prepared selectively from plasma of donors giving a history of overt viral hepatitis two or more years earlier. The proportion of contributors to the globulin who had B hepatitis was unknown but the final product contained a low titer of antibody to the surface  antigen of hepatitis B virus (anti-HBs). The failure of 20 ml of immune serum globulin to reduce the incidence of type B post-transfusion hepatitis (7/93) below that of placebo-treated controls (8/102) was not unexpected in view of the globulin's low titer of anti-HBs. However, more than two thirds of the post-transfusion cases were not type B and were as plentiful among globulin recipients (17/93) as among controls (17/102). Although some of the donors from whom the immune serum globulin was obtained may once have had the same type(s) of hepatitis as the non-B cases currently observed in transfusion recipients, the globulin apparently did not contain enough specific antibody to confer protection in the dose schedule tested.


Role of altered vascular permeability amyloid formation. Heavily radiolabeled plasma proteins were injected intravenously into mice with experimentally induced amyloid lesions, and the distribution of the labeled protein with respect to vascular spaces and the amyloid lesions determined by autoradiography. Radiolabeled albumin and IgG entered early amyloid lesions so quickly and in such high concentration as to indicate that much of the volume of these early lesions must be occupied by circulating plasma. Entry of these same proteins into advanced lesions was definitely reduced. High-molecular-weight proteins remained largely in vascular spaces. In situ saline perfusion resulted in considerable removal of protein from early lesions but very little alteration in their greenish-yellow birefringence when stained with Congo red. An initial alteration in vascular permeability with circulation of plasma proteins in tissue spaces quickly followed by a progressively increasing deposition of fibrous proteins on an underlying cellular framework may be the sequence of events in amyloid formation.


Evidence for an active immune response in acute hyperthyroidism (Graves' disease). Activated lymphocytes, identified by an autoradiographic labeling method, were found to be present in the peripheral blood of the majority of 20 patients with acute hyperthyroidism (Graves' disease). The number of such cells in the blood was significantly greater than that found in 30 healthy controls (p less than 0.00001), and in 13 patients who had previously suffered from acute hyperthyroidism, and who were judged to be euthyroid following therapy (p less than 0.025). This latter group included two patients in whom such activated lymphocytes had been found in the blood during the acute phase of their illness. Furthermore, there were significant differences between the number of such activated circulating lymphocytes in the group of patients with acute hyperthyroidism and five patients suffering from hyperthyroidism due to a toxic thyroid nodule (p less than 0.001), five patients suffering from primary myxedema (p less than 0.001), or in 14 patients with a nontoxic multinodular goiter (p less than 0.05). Identification and counting of circulating T and B lymphocytes by fluorescent immunolabeling and rosette-forming techniques in a small number of the patients with acute hyperthyroidism failed to reveal significant differences from the normal. The results suggest that in acute hyperthyroidism there is active stimulation of the cellular immune system, and that this effect is specific to the early, untreated phase of the disease. This response is different to other thyroid diseases, including hyperthyroidism due to a toxic thyroid nodule.


Hypercalcemia in epidermoid carcinoma of the head and neck. Hypercalcemia occurs rarely at the time of diagnosis in patients found to have epidermoid carcinoma of the head and neck. It is particularly unlike in those patients who present with potentially curable lesions. Only 2 of our 307 patients who presented with potentially curable lesions were hypercalcemic at the time of diagnosis. Two hundred thirty-eight of these patients were followed up for two years or until death. Of the 139 who did not remain free of disease, hypercalcemia of clinical significance developed in ten (7.2 per cent). Pseudohyperparathyroidism was the suspected cause in seven of twelve patients. Debilitating symptoms were present in all patients with serum calcium levels greater thn 12.0 mg/100 ml. Treatment, when given, was effective in alleviating gastrointestinal and central nervous system problems. Although usually temporary, symptomatic improvement was appreciated by most patients and family members.


The dynamics of alpha-fetoprotein and albumin synthesis in human and rat liver during normal ontogeny. The dynamics of synthesis of alpha-fetoprotein (AFP) and albumin have been studied in rat and man by immunocytochemical localization of each protein in the liver and its quantitation in serum at different periods of normal intrauterine and extrauterine life. The proteins are present only in the cytoplasm of hepatocytes. During physiologic development, an inverse relationship exists between the serum concentrations of the two proteins, the latter for each appearing to be directly related to the number of hepatocytes synthesizing it. AFP-containing cells are randomly distributed, while albumin-containing cells are more uniformly spread out. Both, however, are often preferentially located around venous channels. In the case of AFP-containing cells, the last to disappear with advancing postnatal age are seen around the hepatic veins. When both proteins are present in serum, several hepatocytes seem to syntehsize the two simultaneously, though others contain only one of them. It is unlikely that in the physiologic state different populations of hepatocytes are assigned to synthesize AFP and albumin separately.


[Hydroxy-ethyl-starch as plasma substitute in transurethral prostatectomy with the "cold-punch" method (author's transl)]. Each of 24 patients undergoing transurethral prostatectomy under spinal anaesthesia received 1000 ml HES and, depending from the clinical situation, some patients received blood-transfusion. Afterwards two groups were formed: group A (11 patients), who received HES only, and group B (13 Patients) with additional blood-transfusions. The following parameters were monitored simultaneously: blood pressure, heart rate, stroke index, cardiac output, active blood volume, hemoglobin and hematocrit in whole blood as well as in the irrigatin fluid of the bladder. They showed HES to be a useful plasma substitute for older patients. It is well tolerated and has a slow stabilising effect on circulation, which was effective for several hours. Furthermore, HES-infusion reduced the average need of bloodtransfusion by 500 ml.


Orthodontic root resorption studied by electron microscopy. The present study supports previous findings that root resorption takes place simultaneously with and after the elimination of hyalinized tissue. The cementoid layer and the more mature periodontal collagen fibers adjacent to cementum are possible barriers preventing root resorption. The microenvironment around hyalinized tissue is favorable for the introduction of hard-tissue resorbing cells. The findings show that the elimination of hyalinized tissue leads to the removal of the cementoid and the mature collagen thus leaving a raw cemental surface without a barrier. It is therefore hypothesized that such an area is readily attacked by odontoclasts. Once resorption lacunae are established, the cementum is resorbed from the rear as an undermining process. By continued orthodontic force application the resorption process will proceed even after all hyalinized tissue is eliminated. If the orthodontic force is discontinued or falls under a certain level, the resorption lacunae are repaired.


Morphologic resemblance of zygomycete spores to Pneumocystis carinii cysts in tissue. Structures resembling the cyst form of Pneumocystis carinii were noted in the lungs of alloxan-diabetic rabbits to whom spores of several species of fungi belonging to the class zygomycetes had been administered by intranasal instillation. At 24 to 168 hours after the instillation of spores, these rabbits succumbed to diabetes or zygomycete infection, or both, and the possibility of a mixed infection with both fungus and P. carinii could not be ruled out on morphologic criteria alone. To resolve the dilemma, rabbit lung tissue was examined by the fluorescent antibody procedure with labeled P. carinii antibodies. The labeled antibodies did not stain structures that resembled P. carinii but stained P. carinii cells in control rat lung infected with this organism. Investigators should be aware of the difficulty that may arise in differentiating the cyst form of P. carinii from spores of some fungi. The direct fluorescent antibody procedure for P. carinii appears to be very useful for differentiating P. carinii cysts from fungal spores.


[Survival of children and results obtained with a new chemotherapic combination in disseminated neuroblastoma treatment (author's transl)]. A new polichemotherapic combination, including B.C.N.U., is essayed at six patients with disseminated neuroblastoma. Survival of these children and secundaries effects of the drugs are presented.


Uniformity in the clonal repertoire for the immune response to phosphorylcholine in mice. A comparison of the clonal nature of the immune response to phosphorylcholine (PC) was made in nine different inbred mouse strains. Quantitative idiotypic analysis showed that anti-PC antibodies from each strain were composed of antibodies bearing binding-site idiotypic determinants indistinguishable from two different BALB/c myeloma proteins, T15 and M511. Idiotypic determinants of two other PC-binding proteins, M167 and M603, were not detected. Isoelectric focusing of the light (L) chains verified the presence of antibodies similar to T15 and M511 in each strain and indicated the presence of two additional antibodies, one of which has an L chain which cofocuses with M603. Fractionation of anti-PC antibody with anti-idiotypic antibody showed that immunoglobulins bearing T15 and M511 idiotypic determinants are separate and contain L chains that are unifore and resemble those of T15 and M511, respectively. Thus, these mice which differ genetically at multiple loci including the heavy chain allotype complex locus each possess, at least in part, an equivalent set of clonotypes specific for PC. This indicates that the genes encoding these antibodies must be contained in the germ line.


Role of early viral surface antigens in cellular immune response to vaccinia virus. Infection of mice with the vaccinia virus strain WR, Elstree or DIs, a coditional lethal mutant of vaccinia virus, resulted in the generation of vaccinia virus-specific sensitized cytolytic T lymphocytes (CTL). It could be shown by cross-reactivity between the three strains and by inhibition experiments with specific antisera that early vaccinia surface antigens are sufficient for the generation of specific CTL in vivo and for the lysis of infected target cells in vitro.


Isolation of lymphocyte surface antigens. II. Identification of non-beta 2 -microglobulin-associated human T cell-specific antigen. Iodinated cell surface components from human thymus lymphocytes labeled by the lactoperoxidase method, were solubilized by papain digestion and then 3 M KCl extraction of the residual cell pellet. Antiserum to human thymus bound three components from this material, mol. wt. approximately equal to 40 000, 20 000 and 12 000 daltons. This antiserum was absorbed with cultured human lymphoblasts (CHL) until it no longer bound CHL antigens or the HLA-beta 2-microglobulin complex. It continued to bind labeled antigens from thymus, peripheral blood lymphocytes and "T" cell-enriched fraction of tonsil lymphocytes. The absorbed antiserum bound a component from papain-solubilized thymus antigens which had an estimated molecular weight of approximately 40 000 daltons and which was not associated with beta 2-microglobulin. This component seemed to be a human T cell-specific antigen.


Structural and serological relationships among different antibodies from the same rabbit antiserum. II. The idiotypes of ten antibody components from an anti-streptococcal serum. In studies aimed at elucidation of the idiotypic relationships among antibodies produced by a single rabbit, ten antibodies were isolated from the serum of a rabbit immunized with streptococcal Group C vaccine. Four of these antibodies were obtained in amounts sufficient to prepare homologous (rabbit) anti-idiotype antisera. The antisera were used to test for cross-reactivities among all ten antibodies by binding and inhibition of binding assays. Idiotypic cross-reactions among the antibodies were detected by three of the four antisera, while the fourth antiserum reacted only with the antibody that was used as the immunogen for its preparation. Two antibodies demonstrating the highest degree of cross-reactivity had readily distinguishable VH allotypic (a1) subspecificities. The idiotypic reactions and cross-reactions could be inhibited by haptens prepared from the Group C carbohydrate. Bleedings taken from the three immunization periods of this rabbit indicated that the four idiotypes were expressed in approximately equal amounts in each immunization period and that all idiotypes dropped to undetectable levels between immunization periods.


[Sarcoidosis and Whipple's disease. Association? Relation?]. On the basis of a personal case in which two successive diagnoses of sarcoidosis and Whipple's disease were made at an interval of four years, the authors discuss the possible association or relationship between these two disorders, both diffuse, affecting the reticulo-histiocytic system and diagnosed on a histological basis. Review in the literature of 54 cases of Whipple's disease with pleuro-pulmonary lesions makes it possible on the one hand to associate with the latter numerous examples of extra-intestinal involvement, the histological appearance of which of a non-specific epithelioid granuloma does not indicate the diagnosis during the long pre-intestinal phase, and on the other hand to explain that during this period, the presence of pleuro-pulmonary lesions leads to an initial diagnosis of the probability of sarcoidosis. The practical consequences are as follows: The appearance of intestinal signs during "sarcoidosis" should lead to intestinal or mesenteric node biopsy to seek the macrophages with highly PAS positive cytoplasm characteristic of Whipple's disease. Their discovery necessitats long term antibiotic therapy which has recently transformed the outlook in this disease, invariably fatal in the past.


Coeliac plexus block with alcohol for relief of upper abdominal pain due to cancer. One hundred cases of coeliac plexus block in which absolute alcohol was used as a sclerosing agent, performed over a period of 10 years between 1963 and 1973, are analysed. A success rate of 91% in relieving the pain of advanced intra-abdominal malignant disease is claimed and reasons for failures are advanced. The technique described is relatively simple and a 3-day hospital stay is all that is required. It is recommended that this procedure be carried out before the invalid is subjected to repeated injections of narcotic drugs or any form of surgery for pain relief.


[The immunology of psoriasis].

[The immunology of psoriasis]. In recent years, numerous studies have reported the existence of multiple immune disturbances in psoriasis. Estimation of serum immunoglobulins shows an increase in IgA and a possible decrease in IgM. The percentage of psoriatic patients with an increased IgE (450 I.U./ml) is increased in comparison with the normal population. IgA levels in salivary secretions are markedly increased. There are anti-IgG antibodies as well as anti-IgG activity on circulating lymphocytes in 45 p. 100 of psoriasis sufferers. This anti-IgG activity has also been described by certain authors in the psoriatic epidermis. More recently, the abnormal presence of anti-stratum corneum auto-antibodies fixed on the corneal layer has been noted. Cell immunity explored by intra-dermal tests and by the DCNB test is little disturbed. However, certain studies have shown a dissociated deficiency in circulating T-lymphocytes. Finally, it is now well established that BW17, B13 and BW38 histocompatibility antigens are found with a high degree of frequency in psoriasis. The role of these immunological disturbances in psoriatic disease remains to be defined.


Slide staining device for use during space flight. A slide staining device is described that performs Gram and Wright stains during space flight. Reagents and liquid wastes are contained within a closed system.


An electron microscopic technique for the study of elastic tissue in small arteries and arterioles of the kidney. Elastic tissue content and distribution were studied by electron microscopy of small arteries and arterioles of kidneys obtained from normotensive Wistar and spontaneously hypertensive rats, normal dogs and patients with hypertension and glomerular diseases. Specificity and sensitivity of silver tetraphenyl porphyrin sulfonate stain are discussed. This specific electron microscopy technique should prove useful in analysis of elastic tissue in pathologic states of other organs, especially where elastic tissue normally appears to be sparse or absent.


Selective uptake of contact allergens by the Langerhans cell. Using histochemical staining techniques, it has been possible to demonstrate the selective uptake of ten different contact allergens by the Langerhans cells. It is postulated that the Langerhans cells form a reticuloepithelial system that clears the epidermis of foreign chemicals. The Langerhans cell is viewed as the site of hapten binding and antigen formation and hence the central target cell in immune contact dermatitis reactions.


Storage and analysis of samples of water, fish, and mud from environments contaminated with abate. Methods for extraction, cleanup, and analysis of samples of water, mud, and fish containing trace quantities of Abate have been developed. Water was extracted by high-speed stirring of 10 ml of hexane in a 300-ml sample. The extracts were evaporated and analyzed by gas chromatography with a limit of detection of 0.00003 ppm. Dried mud samples were extracted by shaking with acetone. An aliquot of the acetone extract was diluted with water and the Abate extracted into 10 ml of hexane by high-speed stirring. The extracts were analyzed by gas chromatography. Fish were extracted with methylene chloride, cleaned up on a silica gel column, and analyzed by gas chromatography. The limit of sensitivity of the methods for mud and fish was found to be 0.001 ppm. Fish samples were stored for 3 weeks in 10% formalin containing 5% sodium thiosulfate without significant loss of Abate residues. A biological magnification of greater than 100 was observed in fish exposed to Abate for 16 hr at concentrations of 0.02 and 0.002 ppm.


Antithrombin III deficiency causing postsplenectomy mesenteric venous thrombosis coincident with thrombocytopenia. The most commonly recognized cause of mesenteric venous thrombosis following splenectomy is hypercoagulation secondary to reactive thrombocytosis. A case is reviewed in which hypercoagulation followed splenectomy for idiopathic thrombocytopenic purpura (ITP) in spite of persistent thrombocytopenia.  Episodic mesenteric venous occlusion occurred due to antithrombin III deficiency. This hypercoagulable state may be the cause of primary acute mesenteric venous occlusive disease. Symptoms and signs suggesting thrombosis in the portal circulation demand immediate coagulation studies since even in the thrombocytopenic patient thrombotic proglems can occur. Surgical intervention is the treatment of choice for segmental small bowel ischemia; warfarin therapy is indicated when there is evidence of antithrombin III deficiency.


[A new technic for LE cell formation. Preliminary report]. In special designed vials using coverslides glued to segments of polyethilene test tubes of 0.15 ml of fibrin free blood were incubated during 45 min to 37 degrees C in a humid chamber. Blood samples were obtained from patients with disseminated Lupus erythematosus (DLE) and normal subjects (N). Adhered polymorpho-nuclear cells (PNC) to glass were washed with Hank's solution; immediately 0.15 ml of DLE or N serum containing 5X10(6) lymphocytes (L) were added to culture cells and incubated at 37 degrees C during 30 min. Lymphocytes were previously incubated at 37 degrees C during 30 min with either N serum or DLE serum. Thereafter the segment of polyethilene test tube was detached from coverslides and cells attached to glass was washed with Hank's solution and stained with Wright solution. PMN of DLE and N in presence of L of DLE and N incubated with fresh serum of DLE formed 5 to 15% of LE cells. Determining factor for LE cells formation is the serum of DLE. Slides contained only PMN and LE cells which make easy the observation of results. All possible combinations with PMN and DLE serum and N serum allowed inclusion of several negative or positive control groups.


Aldolases of the lactic acid bacteria. Demonstration of immunological relationships among eight genera of Gram positive bacteria using an anti-pediococcal aldolase serum. Reciprocal qualitative and quantitative immunological experiments employing an anti-Pediococcus cerevisiae aldolase serum confirmed many of the interspecific relationships demonstrated previously among lactic acid bacteria with antisera prepared against the Streptococcus faecalis fructose diphosphate aldolase. The extent of immunological relatedness observed between the Lactobacillus and Pediococcus aldolases was markedly gses indicating that the pediococci share closer phylogenetic ties with the rod-shaped lacotbacilli than with their spherical counterparts in the streptococci. In addition to confirming the existence of definitive, but distant, relationships between the lactic acid bacteria and certain gram positive nonsporeforming anaerobes, immunological cross-reactivity was also demonstrated between the pediococcal aldolases and those of Aerococcus viridans.


Immunological comparison of enzymes of the beta-ketoadipate pathway. beta-Carboxy-cis,cis-muconate lactonizing enzyme and gamma-carboxymuconolactone decarboxylase catalyze sequential reactions in the beta-ketoadipate pathway; the subunit sizes of the enzymes from Pseudomonas putida, biotype A, are 40 000 and 13 000, respectively. The cross reaction of antisera prepared against the enzymes was tested with the isofunctional enzymes formed by representatives of other bacterial species. Despite the differences in the subunit sizes of the enzymes, the antisera revealed the same general pattern: cross reaction was observed with the corresponding enzymes formed by other strains in the fluorescent Pseudomonas RNA homology group I and generally was not observed with enzymes from other Pseudomonas species or from other bacterial genera. Exceptions were provided by representatives of Pseudomonas cepacia. Members of this species are classified outside the fluorescent Pseudomonas RNA homology group. Nevertheless, the gamma-carboxymuconolactone decarboxylases from these organisms formed precipitin bands with antisera prepared against the corresponding enzyme from P.putida, biotype A; the lactonizing enzymes from the two species did not appear to cross react. Immunodiffusion experiments with gamma-carboxymuconolactone decarboxylase indicated that a common set of antigenic determinants for the enzyme is conserved among strains that have been classified together by other criteria; the relative immunological distances of the decarboxylases of each taxon from the reference P.putida, biotype A, enzyme were indicated by spurring patterns on Ouchterlony plates. These results suggested that the interspecific transfer of the structural gene for the enzyme is not a common event in Pseudomonas.


Physico-chemical and immunological properties of allophycocyanins. Allophycocyanins were purified from diverse cyanobacteria and one rhodophytan alga (Cyanidium caldarium). The native proteins are trimeric molecules with the structure (alpha beta)3. Representative native allophycocyanins and their alpha and beta subunits were characterized with respect to molecular weight, amino acid composition, isoelectric point, absorption and fluorescence spectra and immunological properties. All of the allophycocyanins studied were strikingly similar with respect to each of these properties. Renatured alpha and beta subunits of allophycocyanin were distinct immunologically from each other, and both cross-reacted with the antiserum to the native protein. Trimeric allophycocyanin was readily reconstituted from the purified alpha and beta subunits. Formation of hybrid allophycocyanins was demonstrated by direct isolation and characterization of the hybrid proteins and by immunological techniques. The results support the view that allophycocyanins are a highly conserved group of proteins.


Cytochemical studies on the localization of methanol oxidase and other oxidases in peroxisomes of methanol-grown Hansenula polymorpha. The localization of methanol oxidase activity in cells of methanol-limited chemostat cultures of the yeast Hansenula polymorpha has been studied with different cytochemical staining techniques. The methods were based on enzymatic or chemical trapping of the hydrogen peroxide produced by the enzyme during aerobic incubations of whole cells in methanol-containing media. The results showed that methanol-dependent hydrogen peroxide production in either fixed or unfixed cells exclusively occurred in peroxisomes, which characteristically develop during growth of this yeast on methanol. Apart from methanol oxidase and catalase, the typical peroxisomal enzymes D-aminoacid oxidase and L-alpha-hydroxyacid oxidase were also found to be located in the peroxisomes. Urate oxidase was not detected in these organelles. Phase-contrast microscopy of living cells revealed the occurrence of peroxisomes which were cubic of form. This unusual shape was also observed in thin sections examined by electron microscopy. The contents of the peroxisomes showed, after various fixation procedures, a completely crystalline or striated substructure. It is suggested that this substructure might represent the in vivo organization structure of the peroxisomal enzymes.


Immunoglobulins in chronic alcoholics: relation to liver histology and effect of 2-month abstinence therapy. Serum immunoglobulin levels and alpha1-fetoprotein were measured in 49 male alcoholics entering an abstinence program. These parameters were evaluated in relation to liver histology. Of 49 studied alcoholics, 32 had elevated serum immunoglobulin levels, chiefly IgA, which occurred in 29 patients. IgA elevation was also observed in patients with normal histological picture of the liver. Of 17 alcoholics with normal immunoglobulin levels, in 9, i.e. 53%, liver histology was normal. In the group of 32 alcoholics with elevated immunoglobulin levles, normal liver histology was encountered only in 8 patients, i.e. 25%. Two-month abstinence in 18 alcoholics with previously elevated IgA levels caused their decrease in 12 patients, i.e. 66%. A decrease in IgA was noted even in patients with symptoms of histological worsening. Of 49 alcoholics, only 1 showed significantly elevated alpha1-fetoprotein.


[Use of automatic devices for histological processing and staining of the tissues and certain characteristics of preparation of histological specimens]. The experience in using automatic devices AT-2, AT-3, AT-4 and in this connection some peculiar features of making histological preparations are discussed. Some modifications of treatment and paraffin embedding of pieces of the tissue are suggested. Recommendations concerning economizing reagents used in the automatic apparatus referred to above without lowering the quality of preparations are given. The technique described makes it possible to prepare specimens and to carry out investigations of bioptic materials in the course of 2-3 days (including the fixation period).


Particles resembling oncornaviruses. Spontaneous release from cultured meningioma cells. A cell culture from thoracic cord meningioma tissue obtained at surgery was maintained for several passages in cell culture. These cells spontaneously released particles with an RNA of high-molecular weight (90S to 95S) and a density similar to that of type C oncornaviruses. The implications of these results and similar findings by others are discussed.


The cell-mediated immune response to ectromelia virus infection. Secondary response in vitro: specificity, nature of effector and responder cells and requirements for induction of antigenic changes in stimulator cells. An in vitro culture method was used to study secondary cell-mediated responses to ectromelia virus infection in mice. Infected, syngeneic spleen cells or peritoneal cells were efficient "stimulator" cells when cultured with "responder" cells obtained from mice infected with ectromelia 4-6 weeks previously. The kinetics of generation of cytotoxic cells in cultures were determined; a peak occurred on days 4-5. A separation procedure performed on the cytotoxic cells showed that activity was associated mainly with the Ig-negative subpopulation (T cell-rich) and that H-2 compatibility between cytotoxic cells and target cells was required. The secondary response was virus-specific, at the level of both induction and target cell lysis, at least so far as ectromelia and lymphocytic choriomeningitis (LCM) viruses are concerned. Seperation of responder cells prior to culture showed that a potent secondary response was generated with the Ig-negative (T cell-rich) subpopulation and only a weak response was observed when the responder cells were Ig-positive (rich in B cells). Infected stimulator cells did not appear to secrete significant amounts of soluble antigen into the medium over 4 days of culture. Thus, antigenic patterns effective in memory T cell stimulation may be largely associated with the surfaces of infected cells.Pretreatment of ectromelia virus with UV- or gamma-irradiation did not impair its ability to induce antigenic changes in stimulator cells. Stimulator cells treated with UV-or gamma-irradiated virus for 1 h and then immediately with pactamycin to inhibit further viral protein synthesis and replication were efficient stimulators, thus indicating that antigenic changes are induced very rapidly on the surface of stimulator cells after uptake of virus. These treatments are being used to further characterize the cellular requirements in the stimulator population.


The specificity of nematode allergens in the diagnosis of human visceral larva migrans. Extracts of nematodes have been used as skin test antigens in the diagnosis of nematode infections for many years. Ascaris lumbricoides and Toxocara canis are two nematodes commonly involved in human parasitism, the latter being associated with the clinical condition of Visceral Larva Migrans. In vitro and in vivo experiments reported in this paper confirm experimentally, as well as clinically, the existence of cross-reacting antigens between T. canis and Ascaris spp., and probably between T. canis and other nematodes. These cross-reactions compromise the usefulness of skin tests in the diagnosis of such parasitic infections.


Comparative immunochemical studies of the serum low-density lipoprotein in several animal species. The antigenic reactivities of the serum LDLs of several groups of animals (mammals, birds, snakes, and fish) have been compared by qualitative and quantitative immunological techniques. The cross-reactivities of the various LDLs were evaluated qualitatively by gel diffusion techniques and quantitatively by the precipitation and inhibition of precipitation tests. The degrees of cross-reaction between human and animal LDLs ranged from 80-88% in the Old World monkeys to 36-58% in the pig, 26-37% in the guinea pig, 1-10% in the chicken, and about 1% in the snakes and fish. These antigenic relationships are consistent with the taxonomic classification of these species, but are relatively closer than those observed for other serum proteins (e.g., albumin, transferrin, and hemoglobin). These results indicate a relatively high conservation of LDL structure through evolution, and are consistent with a common function for this lipoprotein in the transport and metabolism of cholesterol in the higher animals.


[Presence of alpha-2-macroglobulin on the membrane of peripheral human blood lymphocytes : a new lymphocyte maker]. The presence of alpha-2-macroglobulin (alpha-2-M) on the surface of peripheral blood lymphocytes from normal human subjects and from patients with chronic lymphatic leukaemia (CLL) and with ataxia-teleangectasia was studied by indirect immunofluorescence technique. Same experiments were done on purified subpopulations of normal blood peripheral lymphocytes (B and T). The percentage of alpha-2-M bearing lymphocytes is 17 +/- 6% as regard to normal subjects (Ig-bearing cells are 14%): in CLL the percentage of alpha-2-M bearing cells generally is significantly lower than that of Ig-bearing cells (IgM-IgD). On selected subpopulations, 90% of alpha-2-M bearing cells are present among non T-cells and only 5% among T-cells (probably due to not absolute purification). Blocking experiments using anti-Ig sera did not affect significantly the percentage of alpha-2-M bearing cells and using anti-alpha-2-M sera did not affect that of Ig-bearing cells. The percentage of E-rosette forming cells is not affected by pretreatment of peripheral lymphocytes with anti-Ig and/or anti-alpha-2-M sera. Hypothesis is set forth that alpha-2-M bearing cells could be a lymphocytes subpopulation made by a subgroup of B-cells and by K-cells, taking part in that immunoregulatory system in which collaborate many serum alpha-globulins and T-suppresor cells.


[The influence of frequency and blockade of the autonomic nervous system on the functional behaviour of the human conduction system. Part A: Conduction velocity (author's transl)]. 32 patients were studied by His-bundle recordings to examine the extent to which frequency and autonomic nervous system influence the conduction velocity in the subdivisions (atrium, AV-node, His-Purkinje system) of the normal PR-interval. The measurements were performed during sinus rhythm and three electrically induced atrial frequencies before and after intravenous administration of 1 mg Atropine (15 patients) and 0.4 mg Visken (17 patients). In influencing the atrial conduction velocity frequency dominates the blockade of both components of the autonomic nervous system. Increase in frequency lengthens the intraatrial conduction time. Blockade of parasympathicus and sympathicus does not significantly influence the changes in intraatrial conduction velocity induced by increase of frequency. Patients with prolonged intraatrial conduction respond in the same way to cycle length shortening and blockade of the autonomic tone as patients with normal conduction. The results are discussed with respect to acetylcholine and catecholamine influence on the electrophysiological properties of the atrial myocardium. The AV-node is the part of the conduction system most sensitive to the influence of both cycle length shortening and blockade of the autonomic nervous system. Artificially induced cycle length shortening prolongs the intranodal conduction time to a different individual level for each patient. Blockade of the parasympathicus not only shortens this interval but also reduces the steepness of the AH-time induced by atrial pacing. Blockade of the sympathicus has the opposite effect. The most likely explanation for these results is the abolishing of the functional dissociation within the AV-node by blocking the autonomic influence on this structure. The conduction velocity in the His-Purkinje system is influenced neither by atrial pacing nor by blockade of both components of the autonomic nervous system.


[The influence of frequency and blockade of the autonomic nervous system on the functional behaviour of the human conduction system. Part B: Refractory periods (author's transl)]. 32 patients were studied by His-bundle-electrocardiogram and programmed atrial stimulation to examine to which extent frequency and autonomic tone participate in influencing the effective (ERP) and functional (FRP) refractory periods of the atrium and AV-node. The measurements were performed during three electrically induced atrial frequencies before and after intravenous injection of 1 mg Atropine (15 patients) and 0.4 mg Visken (17 patients). For the atrium, frequency dominates the blockade of both components of the autonomic nervous system in influencing both refractory periods. Increase in frequency shortens both ERP and FRP of the atriu. The blockade of parasympathicus and sympathicus does not significantly influence the changes in atrial ERP and FRP induced by atrial pacing. The AV-node responses most sensitive to both pacing induced cycle length shortening and blockade of the autonomic tone. Cycle length shortening prolongs the nodal ERP. the FRP is either shortened or prolonged. Blockade of the parasympathicus shortens both nodal ERP and FRP. Blockade of the sympathicus lengthens both parameters. This behaviour of both refractory periods in response to atrial pacing and blockade of the autonomic tone are discussed with respect to the "gate mechanism" in the conduction system. In the majority of patients blockade of the parasympathicus shifts the "gate" from the AV-node to the atrium. Blockade of the sympathicus has the opposite effect in some cases.


Heterologous specificity of reticulin RS antibodies. Reticulin antibodies were detected by the indirect immunofluorescence technique in 1450 sera from miscellaneous patients. R1 pattern on human and rat tissues was found only in 2 cases with adult coeliac disease. Reticulin Rs staining, involving intrasinusoidal cells of rat liver as well as interstitial and endothelial structures of various rat tissues, was demonstrated in 3.7% of cases, without correlation with gastro-intestinal or skin disorders. Rs pattern, due to IgG antibodies, could be absorbed by rat erythrocytes. Only Rs positive sera showed high titres of anti-rat IgG-type hemoagglutinins. Rat RBC agglutinating activity was demonstrated by a modified indirect Coomb's test. Heterologous Rs antibodies appear unrelated to particular diseases and should not be mistaken for the R1 and R2 patterns, which may be useful for diagnostic procedures.


Antigens in human glioblastomas and meningiomas: Search for tumour and onco-foetal antigens. Estimation of S-100 and GFA protein. Extracts of glioblastomas and meningiomas were analysed by quantitative immunoelectrophoresis for the presence of foetal brain antigens and tumour-associated antigens, and levels of 2 normal brain-specific proteins were also determined. The following antibodies were used: monospecific anti-S-100 (glia specific); monospecific anti-GFA (glial fibrillary acidic protein), (astroglia specific); polyspecific anti-foetal brain (12-16th week of gestation); a polyspecific anti-glioblastoma antiserum, absorbed with insolubilized serum, haemolysate and normal brain extract; polyspecific anti-alpha-foetoprotein; and monospecific anti-ferritin. Using the antibodies raised against the tumours, several antigens not present in foetal or adult normal brain were found in the glioblastomas and the meningiomas. These antigens cross-reacted with antigens present in normal liver and were therefore not tumour-associated. S-100 was found in glioblastomas in approximately one tenth the amount in whole brain homogenate, whereas GFA was found 2-4 times enriched. The 2 proteins were absent in meningiomas. The possible use of the GFA protein as a marker for astroglial neoplasia is discussed. Five foetal antigens were found in foetal brain, but none in the tumours. alpha-Foetoprotein could only be demonstrated in foetal tissue extracts, including foetal brain, but not in tumours. Ferritin was detected in all tumour extracts, although the amounts determined were unrelated to histological tumour type.


Reaction of peptide 89-169 of bovine myelin basic protein with 2-(2-nitrophenylsulfenyl)-3-methyl-3'-bromoindolenine. The C-terminal half of the bovine myelin basic protein, peptide 89-169, was treated with BNPS-skatole [2-(2-nitrophenylsulfenyl)-3-methyl-3'-bromoindolenine], and the products were isolated by repeated gel filtration through Sephadex G-50. They consisted of uncleaved peptide 89-169 in which approximately 30% of the tyrosine had been monobrominated and the tryptophan converted to oxindolealanine, peptide 116-169 modified by partial bromination (30%) of the tyrosine, and two chromatographic forms of peptide 89-115. The major form contained the lactone of dioxindolealanine at the C terminus; the minor form contained the uncyclized oxidation product. Each form of peptide 89-115 was resolved into several components by electrophoresis in polyacrylamide gels (10%, w/w) containing 1 M acetic acid and 8 M urea. The presence of three of these components could be explained by partial deamidation of Asn-91 and Gln-102. Studies on the oxidation of tryptophan-containing model peptides by BNPS-skatole indicated that the reaction can also include partial bromination of the dioxindole and its lactone and partial cleavage at the amino peptide bond of the tryptophan.


Characterization of the apolipoproteins of rat plasma lipoproteins. Purified fractions of three major rat high-density lipoproteins (HDL) and one rat very low-density lipoprotein (VLDL) were isolated by Sephadex gel chromatography or preparative sodium dodecyl sulfate gel electrophoresis. These proteins were characterized by amino acid analysis, end-group analysis, molecular-weight determination, polyacrylamide gel electrophoresis, and circular dichroism. One of these rat proteins, of molecular weight 27 000, appears to be homologous with the human A-I protein. However, rat HDL possesses two additional major components not reported in human HDL - an arginine-rich protein of molecular weight 35 000 and a protein of molecular weight 46 000. The arginine-rich protein of the rat is similar in size and amino acid analysis to the arginine-rich protein reported in human VLDL. A major component of rat VLDL of 35 000 molecular weight appears similar or identical to the arginine-rich protein in rat HDL by every criterion employed for their characterization.


ATP-dependent proton translocation and quenching of 9-aminoacridine fluorescence in inside-out membrane vesicles of a cytochrome-deficient mutant of Escherichia coli,. 1. ATP-dependent proton translocation and ATP-dependent quenching of the fluorescence of 9-aminoacridine were measured in inside-out vesicles derived from a cytochrome-deficient mutant of Escherichia coli. 2. ATP-dependent quenching of fluorescence was inhibited by nigericin gramicidin, NH4Cl, and carbonylcyanide-m-chlorophenylhydrazone. Inhibition was also produced by the ATPase inhibitors N,N'-dicyclohexylcarbodimide (DCCD) and diphenyl phosphorazidate (DPA), and by the respiratory chain inhibitors piericidin A, 2-heptyl-4-hydroxyquinoline N-oxide, and An2+. The inhibition of ATP-dependent fluorescence quenching by the ionophores, uncouplers, and respiratory chain inhibitors was not due to an effect on ATPase activity which was insensitive to these agents. 3. By use of the ATPase inhibitors DCCD and DPA, or by replacing ATP with GTP, ITP and CTP, a correlation between the ATPase activity and the rate of ATP-dependent membrane energization, as measured by fluorescence quenching, was obtained.


The action of a carbonsuboxide dimerized gramicidin A on lipid bilayer membranes. Gramicidin A was dimerized with carbonsuboxide as bifunctional reagent. The effect of the resulting malonyl-bis-desformylgramicidin on lipid bilayer membranes was investigated and compared with the effect of the monomer gramicidin. It was found that the single channel conductance and the ion selectivity are very similar to the behaviour of the monomer molecule, whereas the channel forming kinetics and the life time of the single channel of the malonyl-bis-desformylgramicidin differ strongly from the behaviour of the monomer gramicidin. The electrical relaxations are very small and possibly associated with some structural changes of the membrane after a voltage jump. The single channel lifetime of the malonyl-bis-desformylgramicidin is measured in minutes, whereas for the same lipid system the single channel lifetime in the case of the monomer gramicidin is restricted to 1-2 s. It is concluded that the malonyl-bis-desformylgramicidin-molecule itself (as a single molecule) forms an ionic channel without further association.


Binding of heparin to human antithrombin III as studied by measurements of tryptophan fluorescence. Corrected fluorescence excitation and emission spectra of human antithrombin III have been determined. The fluorescence observed originates almost entirely from tryptophan residues. Reduction of the disulfide bonds followed by carboxymethylation did not change the fluorometric properties of the protein. The binding of heparin to antithrombin III caused a marked fluorescence enhancement by about 30% of the intrinsic protein emission intensity. Various samples of heparin yielded different binding curves. Heparin fractionated by gel filtration seemed to be bound to two sites on antithrombin III with association constants of 0.6-10(6)m-1 and 0.2-10(6)M-1 respectively. Heparin, prepared by affinity chromatography on matrix-bound antithrombin III appeared to be bound to only one site with an association constant of 2.3-10(6)M-1. Under similar conditions heparin caused no increase of the intrinsic protein emission intensity when added to reduced and carboxymethylated antithrombin III. The implications of these findings are discussed.


Immunological relation between 14 S dynein and 30 S dynein from the cilia of Tetrahymena pyriformis. The immunological relation between 14 S dynein and 30 S dynein obtained from Tetrahymena cilia was investigated by using antisera specific for each dynein subunit or some dynein subunits separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Although 14 and 30 S dynein main subunits have different electrophoretic mobilities, our immunodiffusion tests showed that there exists a close immunological relation between them. At least three immunologically different polypeptides designated polypeptides A, B and C are included in the 30 S dynein main band which has been recognized as a single component by electrophoresis, and that the polypeptides designated A',B' and C' are included in the 14 S dynein main bands. Polypeptides A and A',B and B', or C and C' appeared to have a certain common antigenic determinant(s). Polypeptide C of 30 S dynein was shown to possess a certain antigenic determinant(s) specific for 30 S dynein, besides the determinant common with that of polypeptide C' of 14S dynein. The second main component of 30 S dynein proved to be a specific polypeptide of 30 S dynein but not to be a degraded product of the main polypedtide. All antisera reacted with native dynein molecules to some extent, but did not inhibit dynein ATPase (ATP phosphohydrase, EC 3.6.1.3) activity significantly.


Mammalian epidermal keratin: isolation and characterization of the alpha-helical proteins from newborn rat. Neutral buffer-insoluble proteins extracted from newborn rat epidermis with alkaline urea have been purified by chromatography on Sephadex G-150 columns run in the presence of sodium dodecyl sulfate. Two proteins with apparent molecular weights of 60 000 and 68 000, respectively have been isolated and characterized. Spectropolarimetric studies show both of them to be alpha-helical in contrast to the non-helical heavier and lighter species also solubilized with alkaline urea. The amino acid composition of the two proteins, their electrophoretic behavior and their immunological characteristics are essentially identical. Both proteins appear to be major constituents of rat epidermal tonofilaments.


Diphtheria toxin: the effect of nitration and reductive methylation on enzymatic activity and toxicity. Diphtheria toxin is rapidly inactivated upon reaction with tetranitromethane. inactivation is partially prevented in the presence of the substrate NAD. The loss of enzymatic activity and of toxicity is concomitant with the modification of one tyrosyl residue per molecule, located in the fragment A. Completely inactivated toxin (more than 5 nitrotyrosines per molecule) is a good toxin antagonist for HeLa cells binding sites indicating that the integrity of its fragment B is preserved. Methylation of lysyl residues leads to a decrease of toxicity and enzymatic activity but only after the modification of about 20 lysines per molecule. This methylated toxin however can still bind NAD and seems to possess a functional fragment B. Enzymatic site of diphtheria toxin fragment A seems thus to contain one essential tyrosyl residue implicated in the binding of NAD and at least one lysine not implicated in this dinucleotide binding.


The different electrophoretic forms of post gamma-globulin their antigenic identity and their structural variability. Post gamma-globulin, first described as a constant component of protein of cerebrospinal fluid and urine from patients with tubular disorders has also been found in other biological fluids. Post gamma-globulin from a single individual always migrated as several bands after storage. Three electrophoretic forms, immunochemically identical, have been isolated by gel chromatography, preparative continuous flow electrophoresis and ion-exchange chromatography. Their molecular weight was found to be approximately between 11 000 to 12 000. No difference between the three forms could be detected. The N-terminal amino acids were found to be Lys, Arg and Leu respectively for the three forms of post gamma-globulin. The "slow" and "fast" forms of post gamma-globulin seemed to differ by elimination of small basic peptides or amino acids from the N-terminal end of the protein. No enzymatic activity of post gamma-globulin was found, but this requires further investigations.


Cytochemical aspects of Mercenaria mercenaria hemocytes. The hemocytes of the hard clam M. mercenaria were of three types: an agranulocyte, a small, and a large granulocyte. The agranulocyte, with only a thin periphery of cytoplasm surrounding the nucleus, had no visible cytoplasmic granules in living preparations but did exhibit a few centers of nonspecific esterase activity. This cell type represented 2% of the hemocyte population. The small granulocyte possessed four distinct granule types and comprised 61% of the total cell population. Large granulocytes accounted fro 37% of all hemocytes. While they contained the same four granule types identified in the small granulocyte, only one-third the total number were present. The nucleus of all three hemocyte types appeared morphologically similar. The four types of granules observed were a blunt, dot-like, a refractile and a filamentous granule. Blunt granules were identified as mitochondria, based on their ability to reduce Janus Green B to diethyl safranin, the presence of NADH dehydrogenase activity and boundary staining with Sudan black B. Dot-like granules were identified as lysosomes on the basis of neutral red staining, localization of acid phosphatase and nonspecific esterase activity and staining with Sudan black B. Refractile granules were demonstrated to be membrane-bound, lipid-filled structures that reacted positively with Sudan black B and Oil red O, respectively; these granules act as lipid storage centers. Nuclear similarity of the three cell types suggest that these cells might represent different stages of maturity, rather than three distinct cell lines. This was also indicated by the similar yet graded cytochemical reactions and the varying degree of motility and phagocytic activity demonstrated by hemocyte types.


[Effect of chronic stimulation of the negative and positive emotionogenic centers of the hypothalamus on development of cardiac arrhythmias]. Chronic experiments were conducted on 46 rabbits; a study was made of the changes in cardiac activity arising in prolonged (up to 1--2 weeks) electric stimulation of the ventro-median nuclei of the hypothalamus. Disturbances of the cardiac rhythm in the form of tachycardia, atrial flutter and fibrillation, ventricular extrasystole and paroxysmal ventricular tachysystole occurred mostly during the first days of stimulation and ceased after inderal infusion. Functional "weakness" of the cardiac pacemakers was revealed in the animals subjected to 1--2-week stimulation of the ventromedian nuclei. This was indicated by the post-stimulation suppression of the pacemaker automaticity and the appearance of Liciani's periods. Disturbances of the cardiac rhythm originating in stimulation of the ventromedian nuclei ceased in stimulation of the lateral hypothalamic field.


Combination chemotherapy ("CHOP-Bleo") in advanced (non-Hodgkin) malignant lymphoma. Forty-seven adults with advanced malignant lymphoma (the majority in stage IV) were treated with a combination of cyclophosphamide, hydroxyldaunorubicin (Adriamycin), vincristine (Oncovin), prednisone, and bleomycin (CHOP-Bleo). The complete remission (CR) rate was 66%. The overall response (complete + partial remission) was 92%. The CR rate in patients with diffuse histiocytic lymphoma (DHL) was 69%. Only 3 of the 18 patients with DHL in CR have relapsed; the projected median duration of response was calculated to be greater than 2 yr. In patients with nodular poorly differentiated lymphocytic lymphoma (NPDL), the CR rate was 62%. One of the eight patients with NPDL in CR has relapsed; the projected median duration of complete response will be greater than 4 yr. The median survival for all patients entered in this study has not been reached; however, it was estimated that it will be greater than 3 yr. The survival curves became flat at 70 wk for the patients with DHL and at 1 yr for the patients with NPDL. Major complications during chemotherapy with CHOP-Bleo were myelosuppression and alopecia. Only six severe infections occurred during myelosuppression. No hemorrhagic problems were observed. This study indicates that combination chemotherapy with these agents is effective in increasing the CR rate and survival in patients with diffuse histiocytic lymphoma. In patients with NPDL, further observation will be needed to assess the effect of this combination on survival.


T and B lymphocyte antigen-positive null cell leukemias. To clarify the nature of null cell acute lymphoblastic leukemia (ALL) we tested null lymphoblasts and other leukemic cells for human T and B lymphocyte antigens detected by reciprocally absorbed rabbit antisera prepared against autologous T and B lymphoblast cell lines HSB-2 and SB. Blasts from 4 of 4 patients with T cell leukemia and from 2 of 10 patients with null cell leukemia expressed T cell but not B cell antigens. Blasts from 8 of 10 patients with null cell leukemia, one patient with B cell leukemia, and 3 of 3 patients with acute myelogenous leukemia expressed B cell but not T cell antigens. T lymphocyte antigen-positive null cell ALL appear to be a form of T cell ALL. B antigen-positive null cell leukemia could arise from either immature B cell precursors or from multipotential stem cells.


Repair in arterial tissue. A semiquantitative scanning-electron-microscopic and light-microscopic study on the effects of toxic doses of noradrenaline on the endothelium of the rabbit thoracic aorta. The effects of various numbers of intravenous toxic doses of noradrenaline observed at various time intervals after the injections were studied by scanning electron microscopy (SEM) and surface light microscopy of silver-stained rabbit thoracic aorta and by transmission light microscopy. Hexagonal endothelium: a conspicuous change was observed by SEM and surface light microscopy, increased with the length of observation time, and to a certain degree with the number of noradrenaline injections; this change occurred coincidently with development of media necroses, intimal thickening, dilatation of the vessel and various gross changes. Silver-binding endothelial cells: a discrete lesion prevailed in experiments based on one injection of noradrenaline and seemed partly independent of time of observation. The nature of these cells and their role in the morphology of noradrenaline lesion remained obscure. Hexagonal endothelium and silver-binding endothelial cells may have a partial common pathogenetic mechanism, but a mutual relationship was improbable.


A double-blind trial of the effects of candicidin on patients with benign prostatic hypertrophy. 62 patients with benign prostatic hypertrophy, leading to bladder outflow obstruction, took part in a randomised double-blind trial. Those patients in the treatment group took 6 capsules of a polyene macrolide, candicidin, each day (300 mg daily dose). Subjective improvement was noted in both the treated and the control group. This improvement was not confirmed by pressure flow analysis of micturition. The results of this trial do not support the view that candicidin is a useful drug in the treatment of benign prostatic hypertrophy.


Fibreoptic endoscopic palliative intubation of inoperable oesophagogastric neoplasms. Palliative intubation for inoperable malignant strictures at the cardia was done on 16 occasions in 13 patients using fiberoptic endoscopy. Preliminary dilatation was performed, and the Celestin tube was mounted on an introducer and passed over a guide wire inserted with a fiberoptic endoscope. Only one death resulted from the procedure and all the 12 patients who left hospital were swallowing satisfactorily on discharge. The method provided a simple and relatively safe means of relieving dysphagia and improving nutrition.


Concentration of thyroxine-binding globulin: value of direct assay. The concentration of thyroxine-binding globulin (TBG) in the serum can now be measured by direct assays that are simple and inexpensive. Comparison of a direct measurement of TBG concentration with a widely used indirect method (Thyopac-3) showed that the indirect method was inaccurate when TBG concentrations were high. This will result in an increase in the derived free thyroxine index (FTI), so that euthyroid patients with a raised TBG concentration may be at risk of being labelled thyrotoxic. Correction of serum total thyroxine (T4) concentration according to the actual TBG concentration (T4:TBG ratio) provided a better correlation with thyroid state than FTI.


Reutilization of precursor following axonal transport of [3H]proline-labeled protein. In further studies on axonally transported protein in the goldfish visual system, the turnover of rapidly transported [3H]proline-labeled protein was examined. It was found that: (1) a fraction of the rapidly transported protein has a relatively short half-life; (2) [3H]proline released following proteolysis of transported protein is efficiently reutilized for tectal protein synthesis, as inferred from an increased labeling of nuclear protein in the contralateral tectum (COT) relative to that in the ipsilateral tectum (IOT); (3) a small amount of [3H]proline arrives in the COT by axonal flow of the free amino acid; and (4) [3H]leucine and [3H]asparagine are less efficiently reutilized than [3H]proline. These findings may relate to the phenomenon of transneuronal transfer of radioactivity which has been observed with [3H]proline as precursor. The extensive reutilization of [3H]proline may account for part or all of the labeling at secondary synaptic sites. The results suggest that asparagine may be highly suitable for radioautographic identification of primary neuronal fields.


Response of cultured mammalian cells to the exotoxins of Pseudomonas aeruginosa and Corynebacterium diphtheriae: differential cytotoxicity. The sensitivities of 21 mammalian cell lines to the exotoxins of Pseudomonas aeruginosa and Corynebacterium diphtheriae were measured. Each line exhibited 1-4 log differences in sensitivities to the two toxins. No species-specific sensitivities were noted for Pseudomonas exotoxin while diphtheria exotoxin was most potent in cells of monkey origin, followed by human and hamster cells. Rat- and mouse-derived cell lines were very insensitive to diphtheria exotoxin. The rates of cellular intoxication by both toxins exhibited apparent first-order kinetics and were indistinguishable from one another when equipotent doses were used. Our preparation of diphtheria exotoxin appeared to have a slightly higher ADP-ribosylating efficiency than did Pseudomonas toxin. However, neither toxin exhibited cell line-specific differences in ribosylating efficiencies which could have explained the wide range in potencies for intact cells. Our results suggest that there are significant differences in the mechanisms of cellular intoxication by Pseudomonas and diphtheria exotoxins and that these differences probably exist in the attachment or internalization stages of toxin action.


Leukocyte adherence inhibition by soluble tumor antigens in breast cancer patients. The authors modified and refined the Leukocyte Adherence Inhibition Assay (LAI) first described by Halliday, et al. in 1972 by standardizing the protein concentration of tumor-associated antigens (TAA) and by utilizing paired normal tissue extracts as controls to eliminate interference of HL-A histocompatibility antigens and organ-associated antigens. When dose response studies were performed, a progressively larger percentage of patients reacted to the LAI test with increasing concentration of tumor extracts, but the optimal concentration was found to be 200 mug/ml, where 42 out of 66 (63%) leukocytes from 54 breast cancer patients reacted to the breast cancer extracts. At this dose range, only three out of 39 (7%) normal donors and four out of 30 (13%) patients with other types of cancer were positive. When breast cancer patients were tested against TAA of colon cancer and malignant melanoma, one of 24 (4%) and two of 24 (8%), respectively, were positive. Although a higher response rate (72%) was noted in Stage II disease, this was not statistically different from Stage I and Stage III disease. Likewise, no difference was noted in LAI at varying phases following the mastectomy.


The place of radiation therapy in the treatment of non-Hodgkin's lymphomas. The use of Hodgkin's disease as a model for the evaluation and management of the non-Hodgkin's lymphomas may not be appropriate. This latter group of different syndromes and diseases differs significantly in their clinical presentation from each other as well as from Hodgkin's disease. Survival must be separated from relapse-free survival since the latter is a measure of the effectiveness of any individual therapy being applied. Localized nodal lymphoma is uncommon, but important to identify since it is potentially curable by irradiation. Stage I nodular, non-histiocytic lymphomas treated by radiation results in significant, extended, relapse-free survival. All other localized nodal lymphoma is associated with a high proportion of patients relapsing outside the treatment portal. Whole body irradiation is a useful systemic agent causing regression for an extended period of time in stage III or stage IV nodular lymphoma. Chemotherapy seems to have a limited value in nodular lymphomas, with no clear evidence that combination chemotherapy is more effective than single agents. In diffuse lymphomas, aggressive chemotherapy shows more promise, with diffuse histiocytic lymphoma having extended relapse-free survival.


Radiotherapy and rectal cancer.

Radiotherapy and rectal cancer. Studies utilizing preoperative radiotherapy as an adjunct to surgery are reviewed, with results indicating marked increase in survival. Results of postoperative irradiation also appear beneficial, although less extensive data are available. Irradiation for inoperable and recurrent rectal cancer has demonstrated symptomatic relief, particularly avoidance of colostomy, and can render some patients operable and resectable. Adjuvant immunotherapy after irradiation and operation is suggested for further research.


Potentiation of the chemotherapeutic action of bleomycin by combination with inosine on HRS-sarcoma. The chemotherapeutic action of Bleomycin, applied at a dosage which induced only a non-specific tumor inhibition on HRS-sarcoma, was potentiated by combination with inosine. This combination, without increasing toxicity, effected both a significant tumor inhibition and a significant curative action.


Combination chemotherapy with methyl-CCNU (NSC-95441), cyclophosphamide (NSC-26271), vincristine (NSC-67574), methotrexate (NSC-740), and bleomycin (NSC-125066) in advanced bronchogenic carcinoma. Forty consecutive eligible patients with advanced bronchogenic carcinoma and no prior chemotherapy were treated with a 5-drug combination of methyl-CCNU, cyclophosphamide, methotrexate, vincristine, and bleomycin. Of the 36 patients who completed at least one 6-week course of treatment and were considered evaluable, 4 (11%) had partial tumor response. Response by cell type was as follows: 2(14%) of 14 patients with squamous cell carcinoma, 2(18%) of 11 with oat cell carcinoma, and none of 11 with adenocarcinoma. Toxicity in the group of 36 evaluable patients consisted of nausea and vomiting in 24 patients (67%), severe leukopenia (white blood cell count less than 1000 cells/-mm3) in 7 patients (19%), severe thrombocytopenia (platelet count less than 100,000 platelets/mm3) in 14 patients (39%), and bleomycin pulmonary toxicity in 2 patients (6%). This combination does not appear to be more effective than single-agent chemotherapy for bronchogenic carcinoma.


Efficient detection of soft agar grown colonies using a tetrazolium salt. The use of 2-(p-iodophenyl)-3-(p-nitrophenul)-5-phenyl tetrazolium chloride at concentrations of from 0.25-1.0 mg/ml, when added to plates containing soft agar grown colonies, colors the cell clusters a deep brick red against a colorless background. This greatly facilitates the quantitation of such colonies since clusters of cells containing as few as 16-32 cells are visible macroscopically. Incubation for 20 h is optimal although coloration takes place as early as 6-8 h.


Immunological tumour profile: organ-specific carcinoma diagnosis in patients employing the macrophage electrophoretic mobility test. The macrophage electrophoretic mobility (MEM) test provides a highly sensitive in vitro technique for the detection of cell-mediated immunity in man. The principle involved is the lymphokine-mediated reduction of the negative surface charge of guinea pig macrophages shown by the slowing of the macrophages during cell electrophoresis. Lymphocytes from 162 patients were tested by MEM. They were exposed to a battery of KCl extracts from normal and malignant human tissues, to encephalitogenic protein (EP), to carcinoembryonic antigen (CEA), and to thyroglobulin. Variable lymphocyte responses to EP, CEA and KCl extracts from different cancers gave MEM reaction profiles common to patients with carcinomas of the same organ origin.


The influence of antisera specific for alpha-fetoprotein and mouse serum albumin on the viability and protein synthesis of cultured mouse hepatoma cells. The cytotoxic potential of heterologous rabbit antibody directed against mouse serum albumin (MSA) and alpha-fetoprotein (AFP) was investigated in vitro with a cell line (Hepa) derived from the mouse hepatoma BW7756. Anti-AFP in the presence of complement could kill Hepa cells at concentrations of anti-MSA that were virtually nontoxic. The specificity of the anti-AFP was defined by demonstrating that Hepa cell toxicity was dependent upon and paralleled the secretion of AFP in synchronized cultures. Furthermore, neither antiserum could be shown to be significantly toxic to mouse neuroblastoma cells (Neuro-2A). Immunoglobulin purified from pools of antisera was also highly effective in producing cytotoxicity even in a complement-free system. This reaction proceeded more slowly, requiring nearly 48 hr to reach maximum effect in comparison to the 12 hr for complement-mediated toxicity. MSA and AFP are secreted during different phases of the cell cycle. In cultures arrested by isoleucine starvation, labeled AFP appears in the medium 10 hr after release of the blockade in association with S phase. The appearance of labeled MSA is delayed until the first mitosis. Cytotoxic effects of anti-AFP parallel the secretion of AFP in synchronous cultures. Both antisera could be inhibitory to the secretion and synthesis of the proteins of their antigenic specificity. MSA synthesis was more susceptible to this inhibition than was AFP synthesis. The significance of this phenomenon and its association with the differential cytotoxicity of the antiserum are discussed.


The effects of a marginally lipotrope-deficient diet on the hepatic levels of S-adenosylmethionine and on the urinary metabolites of 2-acetylaminofluorene in rats. Hepatic levels of S-adenosylmethionine (AdoMet), of glutathione, and of the microsomal enzymes p-nitroanisole demethylase and benzo(a)pyrene hydroxylase were measured in male and female rats fed a diet marginally deficient in choline and methionine and void of folic acid (lipotrope deficient) or an adequate diet for 0 to 14 weeks with and without added 2-acetylaminofluorene (AAF). The urinary metabolites of AAF were determined throughout the experimental period. After 2 to 4 weeks of dietary administration, the hepatic AdoMet levels were 43% lower in male rats fed the lipotrope-deficient diet than in male rats fed the lipotrope-adequate diet; no differences were found in hepatic AdoMet of females fed the lipotrope-deficient or lipotrope-adequate diets for 2 to 14 weeks. Administration of AAF to lipotrope-deficient female rats for 2 weeks led to a transient decrease in hepatic levels of AdoMet. The administration of AAF for 2 to 14 weeks did not significantly affect hepatic AdoMet in female rats fed the lipotrope-adequate diet or in male rats fed either diet. Female rats fed the lipotrope-deficient diet and treated with AAF excreted decreased proportions of N-hydroxy-2-acetylaminofluorene and increased proportions of 5-hydroxy-2-acetylaminofluorene in their urine. However, the urine of lipotrope-deficient male rats treated with AAF contained increased proportions of N-hydroxy-2-acetylaminofluorene and decreased levels of 5-hydroxy-2-acetylaminofluorene. The urinary excretion of 7-hydroxy-2-acetylaminofluorene by male and female lipotrope-deficient rats treated with AAF was generally similar to that in lipotrope-adequate rats. The lipotrope-deficient diet did not appear to alter the hepatic levels of glutathione, p-nitroanisole demethylase, or benzo(a)pyrene hydroxylase activity was lower in the livers of lipotrope-deficient male rats treated with AAF for 8 to 14 weeks than in the livers of lipotrope-deficient rats not receiving the carcinogen. The altered metabolism of AAF correlated well with the previously reported effects of a marginal lipotrope deficiency on AAF carcinogenesis.


Serum alpha-fetoprotein in a mouse strain (C3H-Avy fB) with spontaneous hepatocellular carcinomas. In a study of the association between circulating alpha-fetoprotein concentrations and spontaneous hepatocellular carcinomas, we examined C3H-Avy fB mice, which with age consistently demonstrate a rapidly increasing incidence of hepatic cancer. Although elevated alpha-fetoprotein levels are observed in association with the majority of these tumors, no elevation of alpha-fetoprotein was observed during the life course of non-tumor-bearing mice despite their age-dependent risk for hepatic cancer. Therefore, whatever the evolutionary or age-related biological changes may be that lead to tumor formation in this mouse, they are not linked to the synthesis of significant amounts of this oncofetal protein.


Immunofluorescent analysis of expression of the RNA tumor virus major glycoprotein, gp71, on surfaces of virus-producing murine and other mammalian species cell lines. The specificity of a single rabbit antiserum pool raised against the purified major glycoprotein, gp71, of Friend murine leukemia virus was determined for a variety of virus-producing mouse, feline, and gibbon ape cell lines by viable cell membrane immunofluorescence absorption. Among murine cells examined, Friend gp71 type specificity was shared only with Rauscher virus-producing cells, and a group specificity was present for all the murine leukemia virus-producing cells tested. Friend and Rauscher murine leukemia virus-infected cells shared interspecies cross-reactivity with feline leukemia and gibbon ape lymphoma virus-producing cells. However, Moloney, Gross, and other virus-producing murine cells shared some, but not all, of these gp71 interspecies determinants with the feline and primate cells. Immunoferritin electron microscopy localized these gp71 antigenic determinants on both virus and cell membranes.


Immunofluorescent analysis of expression of the RNA tumor virus major glycoprotein, gp71, on the surfaces of normal murine cells. The expression of the major glycoprotein, gp71, of murine leukemia virus was studied on the surfaces of a variety of normal murine cell lines with a monospecific rabbit antiserum raised against purified Friend murine leukemia virus gp71. Using viable cell membrane immunofluorescence, most established and early passage normal murine cell lines were significantly reactive with the antiserum, irrespective of neoplastic transformation, strain genotype, or whether they were of embryonic or adult tissue origin. The only murine cells tested not detectably expressing gp71 determinants were BALB/3T3 lines. Although some Friend gp71 interspecies reactivity was discernible on normal murine cells, the principal reactivity was shown to be group specific. Fresh thymocytes from BALB/cJ (GIX-), C57BL/6J (GIX-), and 129/J (GIX+) mouse strains were also reactive with Friend gp71 antiserum, and this activity, as well as that of an antiserum prepared against purified AKR gp71, were also group specific. An activity discriminating GIX+ from GIX- thymocytes was not observed with either Friend or AKR gp71 antisera.


Eight-drug combination chemotherapy (MOPP and ABDV) and local radiotherapy for advanced Hodgkin's Disease. Thirty-seven patients with advanced Hodgkin's disease have been treated for greater than or equal to 3 months with a protocol consisting of alternate monthly courses of MOPP (mechlorethamine, Oncovin [vincristine], procarbazine, and prednisone) and ABDV (adriamycin, bleomycin, DTIC, and vinblastine) with local radiotherapy (RT) to areas of originally bulky disease. This therapy produced CR in 19 of 19 previously untreated patients (100%), eight of nine previously treated with RT (89%), and six of nine previously treated with RT and MOPP (67%). The remaining patients are all PRs tending toward CR status. The median time to CR was 3.0 months. The median time in remission to date for the previously untreated patients is 8+ months (2+-14+). After an induction period of eight cycles of chemotherapy patients are maintained on alternate-month treatment continuing the alternating sequence. During this phase three patients have experienced reappearance of disease (one recurrence, one possible second primary lymphoma, and one recurrence in a patient whose original diagnosis is in doubt). The regimen has been well tolerated. All patients were treated as outpatients. Alopecia and neurotoxicity were mild and myelosuppression was moderate. Clinically significant cardiopulmonary toxicity has been limited to mild radiation pneumonitis in one patient and bleomycin pneumonitis which cleared during prednisone in a second patient.


Combination chemotherapy with three marginally effective agents, CCNU, vincristine, and bleomycin, in the treatment of stage III melanoma. Forty-two patients with stage III melanoma were treated with a combination of CCNU, bleomycin, and vincristine. Seventeen of 35 evaluable patients achieved an objective response (48.5%) including five patients with CRs. The median duration of response was 4 months (range, 1-18 months). The responders had a median survival of 278 days (range, 78-1100 days) and the nonresponders had a median survival of 168 days (range 34-619 days). The toxicity off the combination was within an acceptable range with no toxic deaths.


Phase 1-11 study of DTIC and cyclocytidine in disseminated malignant melanoma. Seventeen patients with disseminated malignant melanoma were treated with DTIC (150 mg/m2, Days 1-5) and cyclocytidine (increasing doses sc, Days 1-10) in a phase I-II study. There was one early death. The remaining 16 patients were evaluable for response and toxicity. Two patients (13%) had CR lasting 8+ and 2+ months, while one patient (6%) had a PR lasting 1 month. Nausea and vomiting was seen in seven patients (44%), jaw pain in four (25%), and orthostatic hypotension in two (13%). Hematologic toxicity was not excessive, nor was it cumulative. The overall response rate of 19% was comparable to that reported with DTIC alone. This drug combination does not appear to offer any therapeutic advantage within the dosage range tested in disseminated malignant melanoma.


Experimental studies on the antiarrhythmic action of a lidocaine analog. The electrophysiologic properties of a lidocaine analog (Kö 1173) was experimentally studied in 41 mongrel dogs. The threshold for occurrence of repetitive ventricular extrasystoles was significantly raised by the administration of 2, 4, and 8 mg/kg of the drug, while atrioventricular and intraventricular conduction times were not affected. After ligation of the anterior descending branch of the left coronary artery, the threshold for occurrence of repetitive ventricular extrasystoles was significantly lowered before the administration of Kö 1173 but it returned to control values after the administration 1 mg/kg of the drug. The strength-interval curve shifted profressively to the right when the dose was increased from 2 to 4 and 8 mg/kg. These results indicate that Kö 1173 prevents the decrease in threshold for occurrence of repetitive ventricular extrasystoles in acute coronary insufficiency at a dose which does not affect the conduction system.


"Gomori-positive" neurosecretion in the rat after deafferentation of the medial basal hypothalamus. A portion of the "Gomori-positive" peptidergic neurosecretory (NS) cells in the paraventricular and especially in the supraoptic and postoptic nuclei degenerate three weeks after deafferentation of the medial basal hypothalamus. Most of the remaining NS cells show signs of high activity. Regenerating NS fibres form "muffs" around the blood vessels laterally from the lesion; some of them enter the "isolated" area or persist there if a thin layer of the brain tissue is left somewhere untouched under the basal end of the cut. The regenerating NS fibres are also found outside the nervous tissue: within the scar tissue, in the proliferating connective tissue of the brain sheet below the basal end of the cut and in the mantel plexus area. The NS fibres make close contact with blood vessels invading or penetrating the vascular wall. It is suggested that peptide neurohormones discharged from the "Gomori-positive" NS terminals enter the general blood circulation as well as the portal blood at the site of these newly formed axovasal contacts. It is supposed that under these conditions monoaminergic terminals do not discharge monoamines because no stimulation of monoamine-producing NS cells occurs with deafferentation.


Ultrastructural studies on the secretory cycle of the neurosecretory cells and the formation of Herring bodies in the paraventricular nucleus of the rat. The ultrastructure of the neurosecretory cells in the paraventricular nucleus of the normal male rat was studied by electron microscopy during various functional states. Four morphologically distinct types of neurosecretory cells were observed. It appears that they do not represent different classes of cells but different phases of secretory activity of a single cell type. The perikarya of the neurosecretory cells show a definite cycle of formation and transportation of secretory granules. We have designated the phases of this cycle as: (1) phase of synthesis, (2) phase of granule production, (3) phase of granule storage and (4) phase of granule transport. The neurosecretory granules appear to be moved in bulk into the axons, forming a large axonal swelling filled with granules as a result of one cycle in the neurosecretory process. Thus it may be postulated that a secretory cycle in the perikaryon of the neurosecretory cell seems to result in the formation of a Herring body in its axon, and that its content is then conveyed to the posterior pituitary.


Further evidence for the existence of intralobular nerves in the rat liver. In a recent study (Skaaring and Bierring, 1976) we found cholinesterase-positive nerve-like structures in the lobules of rat liver, and scanning electron microscopy revealed cords having a distribution pattern similar to that of the cholinesterase-positive structures. To obtain further evidence for an intralobular nerve supply the methods of cobalt and Procion Yellow nerve staining (Stretton and Kravitz, 1968; Iles and Mulloney, 1971; Pitman, Tweedle and Cohen, 1972) were adapted, iontophoretic introduction of the dyes being attempted through cut axonal ends in the surface of small excised blocks of rat liver.


Cells transformed by certain strains of Moloney sarcoma virus contain murine p60. It was previously demonstrated that the 60,000 dalton (p60) precursor-like polyprotein containing murine p30 was a constituent of the feline leukemia virus pseudotype of Moloney sarcoma virus [m1MSV(FeLV)]. It is now shown that p60 is detected  in cells of five mammalian species transformed by m1MSV, indicating that p60 is specified by this genome. Moreover, little or no murine p30 is detected in the m1MSV-transformed cells, suggesting that the murine group p30 antigenic reactivity of S + L- cells is ude to p60. Pulse-chase studies in cells producing m1MSV(FeLV) show that p60 is the largest polypeptide detectable during the pulse, and that intracellular p60 is not cleaved into smaller (for example, p30) polypeptides during chase periods of up to 10 hr. The lack of cleavage of p60 is in contrast to the properties of p30 precursors detected in cells containing replicating avian or mammalian RNA tumor viruses. The inefficient cleavage of intracellular p60 and the kinetics of appearance of murine p30 in extracellular m1MSV(FeLV) suggest that p60 cleavage to p30 occurs in cells shortly before virus release. While only p60 was detected in the m1MSV-transformed cells, p60 and p70 were detected in m3MSV-transformed cells, and no immunoprecipitable polypeptides were detected in HT-1 MSV-transformed cells. The observed differences in the intracellular polypeptide expression by each of the strains of MSV suggests differences in genetic content.


The effect of interferon on de novo infection of Moloney murine leukemia virus. The mode of action of interferon in de novo Moloney murine leukemia virus (Mo-MuLV) infection of mouse bone marrow/thymus (TB) cells was studied. Our results indicate that in interferon-treated cells, there is approximately a 2000 fold decrease in the production of infectious MuLV, but only a 10-20 fold decrease in the level of viral specific extracellular reverse transcriptase activity, and only about a 2 fold difference in the number of virus particles observed on the cell membrane as determined by scanning electron microscopic (SEM) studies. Transmission electron microscopic (TEM) studies showed that the proportion of early budding virions, which have shallow crescent-shaped ribonucleoprotein cores (Figure 3A), to virions in later stages of assembly (Figures 3B-3D) is relatively higher in interferon-treated cells than in the untreated controls. From a temperature shift-down experiment on a temperature-sensitive mutant of MuLV, ts 3, which produces viral particles that fail to dissociate from the cell surface at the nonpermissive temperature, we demonstrated that ts 3 virions partially assembled on the cell membrane prior to the addition of interferon are able to complete assembly and to dissociate from the cell membrane on temperature shift-down in the presence of interferon action. Our data suggest that interferon neither inhibits the late stages of virion assembly at which ts 3 virions are arrested at the nonpermissive temperature nor prevents release of the virions. Our findings also indicate that in interferon-treated cells, most of the extracellular virions are noninfectious.


[Presence in immunostimulated cells of an RNA molecule utilizable as template for reverse transcriptase of avian myeloblastosis virus (AMV)]. Cytoplasmic RNA extracted from antigen stimulated immunocompetant cells is transcribed in vitro into DNA by the RNA directed DNA polymerase from avian myeloblastosis virus, in the absence of any added primer. Cytoplasmic RNA from other organs of the same animal, from non-stimulated immunocompetent cells, or from cells in tissue culture is not transcribed in the absence of exogenous primer.