Cross-reactions between Haemophilus influenzae and nineteen other bacterial species. Forty-one different antigens were demonstrated in an antigen preparation obtained by sonication of a Haemophilus influenzae (H. influenzae) type b strain, using crossed immunoelectrophoresis and antiserum obtained from rabbits. Antigens were characterized by absorption experiments with whole heat-killed bacteria, temperature resistance and protein and polysaccharide staining. Cross-reactions between H. influenzae type b and 19 other bacterial species were studied by various quantitative immunoelectrophoretic methods, using the reference system. A non-capsulated H. influenzae cross-reacted extensively (41 antigens) with H. influenzae type b and Haemophilus parainfluenzae, and Haemophilus haemolyticus showed considerable cross-reactivity with H. influenzae type b (26 and 32 antigens respectively), while antigens from eight other bacterial species cross-reacted to varying degrees with one to five H. influenzae antigens.


O-antigenic cross-reactivity in Fusobacterium nucleatum. Lipopolysaccharides (LPS) of the same chemotype isolated from seven strains of Fusobacterium nucleatum were examined by indirect haemagglutination and inhibition of haemagglutination in unabsorbed and absorbed antisera. Four common major antigenic specificities were detected, and two of the LPS examined contained antigenic specificities not shared with any of the other LPS. In addition, a few weak cross-reactions were observed, also with some LPS of other chemotypes.


Chloride transport by self-exchange and by KCl salt diffusion in gramicidin-treated red blood cells. The permeability of gramicidin-treated human red blood cell membranes to K+ and Cl- has been measured at normal ionic strength (1) by tracer exchange at steady-state distribution of salt, and (2) by net transport of salt in the presence of a salt concentration gradient. Under both conditions KCl was the only inorganic salt in cells and medium. In the studies of self-exchanges the electrical driving force on the ions was zero. Calculaton of permeability coefficients from net salt transport was simplified because the experiment was designed as a special case of the Nerst-Planck diffusion regime, i.e. the single salt case. Gramicidin altered the cell membranes from being anion to become cation selective. Gramicidin increased the potassium exchange without affecting the chloride exchange measurably. The chloride exchange showed saturation kinetics as does chloride exchange in normal cells. The net transport of KCl in the presence of a constant concentration gradient increased to a constant value with increasing gramicidin concentration. At high gramicidin concentrations (0 degree C, pH 7.2) the "chloride permeability coefficient" calculated from tracer exchange (1.9 x 10(-6) cm/s) was 290 times the chloride permeability coefficient calculated from net salt transport (0.65 x 10(-8) cm/s). The latter value corresponds to a chloride conductance of 4.2 x 10(-6) ohm-1 cm-2. The chloride permeability coefficient was 2.1 x 10(-6) cm/s at 25 degrees C (pH 6.8) indicating a value of 3 for the Q25. It appears that normal red cells are anion selective in the sense that anion permeability exceeds cation permeability with a factor of more than a hundred between 0 degrees C and body temperature. The anion exchange, i.e. the Hamburger shift, is a tightly coupled transport process which is several orders of magnitude faster than anion transport by salt diffusion.


Immuno-chemotherapy of malignant lymphoma using OK-432, a streptococcal agent. Clinical trials of immuno-chemotherapy were conducted on malignant lymphoma patients. Patients during the period from 1972 through 1977 were allocated to two groups retrospectively according to the mode of treatment, i.e., chemotherapy alone (historical control group, 35 patients) and chemotherapy with OK-432 (treated group, 15 patients). Comparisons were made of the two groups, which were homogeneous with regard to induction chemotherapy, maintenance chemotherapy, stage and histologic type of disease. The treated group had a higher remission rate, and a longer remission duration and survival than the control groups, especially in patients with Hodgkin's disease but the difference was not statistically significant owing to the limited number of cases.


Heat-stable somatic antigens of a group of unclassified fluorescent pseudomonads (UFP). (i) Isolates belonging to a group of unclassified fluorescent pseudomonads (UFP) are similar to Pseudomonas aeruginosa not only in cultural behaviour but also in basic serological properties of their somatic antigens. Living bacteria and cultures subjected to prolonged heating at 100 degrees C or above agglutinated readily in homologous O serum, whereas after exposure to 60 degrees C, ethanol, saturated sodium chloride and formalin the cells of both species became practically inagglutinable. Surface factors inhibiting the agglutination of living bacteria in O sera being absent, the slide technique was chosen as a routine method for the serological grouping of UFP. (ii) The O antigens of UFP were different in serological specificity from those of P. aeruginosa: the two organisms were related only by minor "common" antigens detectable with bacteria heated at 130 degrees C. One hundred and ten out of 115 UFP isolates were classified into 17 serological groups; O groups 2, 5, 10, 12 and 16 were each further divided into two subgroups. Five isolates reacted in several sera or were unstable. (iii) Serological grouping of UFP is reproducible and adequate for the tracing of isolates and may be helpful for a rapid differentiation of the organism from P. aeruginosa.


Subpopulations of human peripheral blood cells: analysis of granulocytic progenitor cells by flow cytometry and immunologic surface markers. Normal human peripheral blood cells were separated into different populations based upon isopycnic sedimentation, E rosetting, and EAC rosetting. Each population was characterized according to morphology, surface markers, granulocytic colony formation in semi-solid media, and stainable RNA content by acridine orange (AO) flow cytometry. These techniques enrich for a population of cells that is characterized by a lymphoid morphology, a high granulocytic-macrophage progenitor cell cloning efficiency, a lack of surface markers, and a high stainable RNA content not found in the other two populations of peripheral blood lymphocytes (T cells and B cells). The stainable RNA content serves as a new metabolic marker for the population of cells in which the preponderance of granulocytic progenitor cells reside.


[Histochemical studies on the structure and the age changes of Descemet's membrane in cattle (author's transl)]. Descemet's membrane of 15 cattle-corneas, aged 6 month to 13 years was studied by various histochemical and morphometric methods. 3 zones could be differentiated. Zone 1 is relatively small and does not show significant changes with age; zone 2 comprises the main part of Descemet's membrane, its diameter increased with age. Zone 2 is rich in sulfated glycosaminoglycans. Zone 3, below the endothelial lining, does not develop before the 3rd and 5th year of life and stains different from zone 2.


[Effect of pyridoxine on histamine liberation and degranulation of rat mast cells]. Pyridoxine, one of the B vitamins, has been shown to be useful in the treatment of childhood bronchial asthma by Collip et al. (1975). A double-blind study with 76 asthmatic children followed for five months indicated significant improvement in asthma following pyridoxine therapy (200 mg daily) and a reduction in dosage of bronchodilators and cortisone. Other reports have shown that nicotinamide, another B vitamin shows inhibitory activity in rat mast cell degranulation and histamine release (Bekier et al. 1974, Wiczolkowska and Maslinski, 1975, 1976). These results induced us to investigate if pyridoxine, like nicotinamide or disodium cromoglycate, exhibits pharmacological inhibitory activity in rat mast cell degranulation and histamine release induced by antigen or other non-immunological stimulants. We found that pyridoxine at concentrations of 10 (-3) M, or greater significantly inhibited rat mast cell degranulation and histamine release induced by phospholipase A, compound 48/80, antigen (egg albumin) or a mixture of dextran and phosphatidyl serine, respectively. In these experimental models, pyridoxine shows a pharmacological profile similar to nicotinamide and disodium cromoglycate, although weaker than the latter. In spite of this, the lack of toxicity of this vitamin at relatively high doses (1 or 1.5 g), the possibility that other mechanisms of action may be involved, such as the improvement in tryptophan metabolism reported by Collip following pyridoxine therapy, suggest that this vitamine merits additional research.


Mycoplasma pneumonia presenting as meningoencephalitis and hemolytic anemia. Simultaneous occurrence of cold agglutinin hemolytic anemia and central nervous system disease due to Mycoplasma pneumonia is extremely rare. A patient is described who initially presented with severe hemolysis and diffuse neurologic dysfunction which appeared during the period of roentgenographic resolution of her pneumonia. Spectrum, pathogenesis, and antimicrobial treatment of the neurologic complications are reviewed.


[Specific soluble cell-wall antigens of "Candida tropicalis" demonstrated by crossed immunoelectrophoresis (author's transl)]. In the present study, soluble cytoplasmic extracts of Candida albicans and C. tropicalis were prepared and showed respectively 67 and 47 precipitates in crossed immunoelectrophoresis performed with a commerical goat anti-C. albicans immunserum. Purified cell walls of C. tropicalis have been prepared, and corresponding antiserum was got in rabbits. Although important cross-reacting antigens exist between soluble cytoplasmic extracts of C. albicans and C. tropicalis, the C. tropicalis cell wall antiserum precipitated antigens in crossed immunoelectrophoresis only in C. tropicalis soluble cytoplasmic extracts. The present study: - confirms that C. albicans shared 70% of the antigens with C. tropicalis; - demonstrates that soluble cytoplasmic antigens prepared by homogenization of yeasts contain cell wall constituents; - allows us to prepare a specific immune serum for C. tropicalis.


C- and G-banding patterns and chromosomal morphology of some breeds of Australian cattle. A cytogenetical study using metaphase chromosomes from cultured lymphocytes, was made of 2 Banteng (Bibos banteng) steers and 218 bulls representing 13 purebreeds (Bos taurus type, Bos indicus type and Sanga) and 7 cross-breeds. Studies were made of photographic karyotypes of Giemsa stained and C-banded chromosomes of bulls of each breed and of B-banded chromosomes from 3 breeds of Bos indicus and one cross-breed Australian Friesian Sahiwal) cattle. The relative lengths of chromosomes of Bos taurus and Bos indicus bulls were compared and significant difference in relative lengths of the X chromosomes were noted between these two species. There was a differences in morphology of the Y chromosomes; Sanga, Banteng and Bos taurus type breeds had a small submetacentric Y chromosome, except for the Jersey which had a metacentric Y chromosome. All Bos indicus type bulls had an acrocentric Y chromosome but the Droughtmaster breed had two forms of the Y chromosome (submetacentric and acrocentric). The C-banding patterns of the autosomes and X chromosomes were similar for all breeds while those of the Y chromosomes of Bos indicus type cattle allowed their accurate identification. G-banding patterns of Bos indicus resembled those of Bos taurus and enabled pairing of homologous chromosomes. Centromeres of the autosomes were unstained but those of the sex chromosomes were darkly stained.


Efficiency and safety of methidathion applied as a pour-on systemic insecticide for control of cattle lice. Methidathion applied to cattle as a pour-on insecticide for control of lice (predominantly Linognathus vituli, but also Haematopinus eurysternus and Damalinia bovis) caused a reduction in the lice population of approximately 98% to 99% in laboratory trials at the minimum recommended dose of 4 mg/kg and a 98.8% to 100% efficiency was achieved in field trials. In a comparative efficiency trial in the laboratory methidathion at 3.5 to 5 mg/kg reduced the lice population by 98.8% to 99%, fenthion by 98.5% at 4.5 mg/kg and famphur by 99.7% at 16.5 mg/kg. Methidathion was tolerated by calves aged 15 to 20 weeks at dose rates up to 40 mg/kg indicating an approximate 7 fold safety margin, but 1 of 4 calves treated at 50 mg/kg died following treatment. Treatment with fenthion at 50 mg/kg, 7.4 times the average recommended rate, famphur at 75 mg/kg, 3 times the average rate and chlorpyrifos at 85 mg/kg, 5 times average rate, caused reductions in whole blood cholinesterase activity of 52%, 27% and 47% respectively which were similar to the reductions in cholinesterase activity found in calves treated with methidathion at similar levels above the recommended commercial dose rates. It was found that 2 day old calves were more sensitive to treatment with methidathion than calves 9 or 16 days old. A further 11,900 cattle of varying age, breed and sex were treated with methidathion under field conditions at the recommended rate of 4 to 8 mg/kg, and 534 cattle were treated at 24 mg/kg without any signs of toxicity.


Genetic regulation of CFA expression in interspecific avian hybrids and somatic cell hybrids. Chicken fetal-leukemic antigen (CFA) is an oncodevelopmental antigen present on embryonic and neonatal chicken peripheral red blood cells (RBCs) but is not restricted to fetal stages of development in other avian species. Crosses between white Leghorn chickens and Japanese quail resulted in adult hybrids whose peripheral RBCs were positive for CFA. Of the four CFA determinants normally found in adult quail RBCs, only two were present on quail-chicken hybrid RBCs. Adult quail--chicken hybrid RBCs also possessed on CFA determinant associated with early development in both quail and chicken and one chicken-specific CFA determinant. Evidence is presented for the possible association of CFA-positive adult peripheral RBCs and the level of circulating reticulocytes. Crosses between pheasant and turkey (both with CFA-positive adult RBCs) resulted in hybrid adult RBCs expressing only a portion of the parental CFA determinants. Through the formation of somatic cell hybrids between adult chicken and embryonic Japanese quail RBCs, it was possible to induce the appearance of CFA determinants normally restricted to embryonic chicken RBCs. Approximately 50% of the hybrid cells showed reexpression of CFA, and this induction was both time and temperature dependent. Hybridization between RBCs of adult chicken and those of either adult Japanese quail or adult turkey failed to elicit the reexpression of chicken-specific CFA.


Biochemical and immunological studies of three genetic variants of 3-phosphoglycerate kinase 2 from the mouse. Three electrophoretic variants of 3-phosphoglycerate kinase 2 (PGK-2A,PGK-2B, and PGK-2C) were purified from DBA/2J, C3H/HeJ, and C57L/J mice, respectively. PGK-2C exhibits only 2% of the specific activity of PGK-2A and PGK-2B in the reaction leading to the formation of 1,3-diphosphoglycerate. Compared to PGK-2A and PGK-2B, PGK-2C exhibits broader coenzyme specificity and lower Kms for substrate and coenzymes.  Incubation at 45C revealed immunionactivation and double immunodiffusion studies showed that mice carrying any one of these three PGK-2 alleles have similar amounts of proteins for PGK-1 and PGK-2 in testes.  The results of these studies suggest that low PGK-2C activity in C57L/J mice is a result of a structural rather than a regulatory gene mutation.


Genetic variation in mouse salivary amylase rate of synthesis. Heterozygotes from matings of the mouse strains YBR/Cv and C3H/As have about 3 times more YBR-amylase than C3H-amylase in the saliva.  The determinant for this quantitative effect is located on linkage group XVI close to or with in the structural gene for salivary amylase.  The quantitative effect is the result of an increase in the rate of synthesis of YBR-amylase, and the determinant is cis acting.  Studies of other mouse strains suggest that regulatory genetic elements may modulate salivary amylase production.


Phosphorescence of protein-bound eosin and erythrosin. A possible probe for measurements of slow rotational mobility. We used a pulsed dye laser working at 540 nm to excite triplet-state formation of eosin and erythrosin, either bound or unbound to bovine serum albumin, in aqueous solution anaerobically at pH 8 and 20-22 degrees C. Delayed emission from radiative transitions of the triplet state was readily detectable, both as delayed fluorescence and as red phosphorescence. Detection of the triplet state by measurement of phosphorescence at 645 nm upwards was at least 100-fold more sensitive than by absorbance measurements of ground-state depletion at 500 nm. When immobilized in poly(methyl methacrylate), the phosphorescence of eosin and erythrosin was polarized with an anisotropy parameter [Jablonski (1961) Z. Naturforsch. A16, 1-4] of about 0.25. The phosphorescence of erythrosin is sufficiently intense to be distinguishable from the long-wavelength end of fluorescence under conditions of continuous rather than pulsed excitation. Our observations suggest that phosphorescence depolarization of eosin or erythrosin probes could be used as a highly sensitive method of measuring rotational relaxation times in region from 10(-5) to 10(-3) s, such as those of the uniaxial rotation of membrane proteins.


Antiarrhythmic effectiveness of creatinol O-phosphate in man. Antiarrhythmic effectiveness of N-methyl-N-(beta-hydroxyethyl) guanidine O-phosphate (creatinol O-phosphate, COP) has been investigated in 10 patients with ischemic heart disease and frequent premature ventricular contractions (PVCs). Each patient received a random succession of treatment with the drug (1020 mg) and with a reference substance (solvent of COP) both administered i.m. twice a day over a 3-day period. In each patient a Holter ECG was recorded in the basal state and during the last 24 h of each treatment. Heart rate, PQ and QTc showed no changes. On the contrary, PVCs were significantly lower (P less than 0.01) with COP than with the reference substance. The drug showed a prevailing effect during daytime. Results are discussed in view of experimental observations suggesting that COP has a "membrane effect".


[Expression of viral antigens on the membrane of normal and leukemic thymocytes of AKR mice]. Expression of antigenous determinants of structural proteins G-MuLV (p10, p12, p30, gp14, gp17) and R-MuLV (gp69/71, p15) on thymocytes of normal and leukemic AKR mice was studied by membrane immunofluorescence. From this sign sharp difference between normal and malignant thymocytes was shown. A possible role in the antitumour immunity of antigens to structural viral proteins MuLV expressed on the leukemic cell membrane is discussed.


[Differences in the properties of antibodies and natural antiglobulin-homoreactants revealed by the method of human IgG chemical modification]. The naturally occurring antiglobulin factors - homoreactants, contained by human IgG preparations, are inactivated as a result of incubation in a solution of sodium rhodanide (3 - 5 M) and sodium desoxycholate (0.005M). Staphylococcal antitoxin contained by the same human IgG preparations is resistant to the action of the reagents. The data obtained indicate the differences in the structure of homoreactants and antibodies.


[Ketotifen, a selective histamine liberator]. Ketotifen-induced histamine release from isolated rat mast cells is inhibited under conditions of ATP content depletion in the mast cells. The histamine release is a temperature-dependent process and is blocked at a low temperature. It is concluded that ketotifen is a selective histamine liberator. Time course of ketotifen-induced histamine release corresponds with the process of partial cellular uptake of histamine released.


[Intensity of RNA and protein synthesis in fibroblasts of mice in wound healing]. The RNA and protein synthesis in mouse fibroblasts was studied on the 3d and 7th day of wound healing by electron-microscopic autoradiography with double-labeling technique. It was shown that the ratio between the protein synthesis in cytoplasm and the RNA synthesis in the nucleus of some cells significantly differed resulting in relative predominance of one of the two processes studied in the fibroblasts during both terms of wound healing. On an average the fibroblasts were characterized by a higher level of the protein synthesis on the 3d day of wound healing. This average high level was provided by an increase in the fibroblasts, in which the protein synthesis prevailed over the RNA one.


[Effect of embryonal organ-specific RNA on myocardial hypertrophy]. The relative weight of the heart and the thickness of muscular fibers in the left ventricular myocardium of rats were evaluated 70-91 and 120-227 days after creating stenosis of the abdominal portion of the aorta. Half of the animals were inoculated with cardiac type-specific embryonal RNA, the rest of the animals being intact. The data were treated by the variational statistical technique, by dispersion and mathematic analyses. Administration of RNA decreased the rate of myocardial hypertrophy development.


Use of antisera against bovine (NCDV) and simian (SA11) rotaviruses in ELISA to detect different types of human rotavirus. Two ELISA systems for the detection of human rotaviruses were developed. In the first system antibodies to Nebraska calf diarrhea virus (NCDV) were used for coating the solid matrix and for the preparation of the enzyme conjugate. In the second system antibodies to human rotavirus and antibodies to simian rotavirus (SA11) were used for coating the solid matrix and for the preparation of the enzyme conjugate respectively. The second ELISA system proved to have a broader spectrum for the detection of human rotaviruses. By using the two ELISA systems, the different types of human rotavirus could be distinguished. The ELISA tests developed were 8 to 64 times as sensitive as electron microscopy (EM) and (or) counterimmunoelectrophoresis (CIEP). The antigen detected by ELISA was shown to be different from that detected by the hemagglutination test.


The kallikrein-kinin system in blood pressure homeostasis. 1. The acute effects of the kallikrein inhibitor aprotinin (498 ki.u./min), and the kininase II inhibitor SQ 14,225 (250 MICROGRAM), GIVEN INTRAVEnously during saralasin-induced angiotensin blockade, were studied in conscious sham-operated rats and rats with benign and malignant two-kidney, one-clip Goldblatt hypertension during dietary sodium restriction. 2. The blood pressure of conscious sham-operated rats increased significantly in response to aprotinin. It remained unchanged after SQ 14,225 in contrast to the significant vasodepressor effect seen when SQ 14,225 was given to the same rats under surgical stress and pentobarbital anaesthesia. 3. Benignly hypertensive rats showed a consistent vasopressor response to aprotinin and a marked vasodepressor response to SQ 14,225. The effects of both inhibitors were markedly and significantly blunted in malignantly hypertensive rats. 4. Our demonstration that two agents with known opposite actions on the kallikrein-kinin system produced predictable and opposite effects on blood pressure may indicate that this system is involved in the homeostatic regulation of blood pressure. It may play an important antihypertensive role in benign two-kidney, one-clip Goldblatt hypertension, a role which might be impaired in malignant hypertension.


Differentiation antigens of the lymphocyte cell surface. Many alloantigens and xenoantigens of lymphocytes are not found generally on other tissues, and this suggests that much of the lymphocyte cell surface is differentiated in comparison with other cell membranes. These differentiation antigens are probably molecules that mediate lymphocyte-specific functions, and are also of interest in that they provide markers for different lymphoid cell types and may be important as target antigens for immunosuppressive anti-(lymphocyte) sera. The purification of differentiation antigens will be important in allowing their molecular properties to be discovered, and will also lead to the production of strong, specific antisera that can be used in functional studies. Radioimmunoassays have been developed for the analysis of anti-(lymphocyte) sera, and these assays provide advantages in purification studies over other techniques. The features of these assays are discussed, and studies of differentiation antigens of rat lymphocytes are described. These include the purification and characterization of the rat Thy-1 antigen, and preliminary studies on two other rat lymphocyte differentiation antigens.


The morphology of normal and denatured polytene chromosomes from Drosophila melanogaster. Normal and denatured polytene chromosomes from Drosophila were compared by light, transmission (TEM) and scanning (SEM) electron microscopy, under different conditions of pretreatment, fixation and specimen preparation. Apart from variations in contrast, acid-alcohol squashes of normal chromosomes were quite comparable in terms of resolution and image quality, regardless of mode of visualization, i.e. stained, unstained, phase contrast, etc. These chromosomes do not appear markedly different from the unfixed or "native" chromosomes isolated by other workers. Thus at the light microscope level at least, any soluble components extracted by acid-alcohol fixation seem to play minor structural roles only. SEM examination of similar squash preparations provided useful topographic views of selected chromosome regions, particularly in favourable spreads where metal coating and tilt angles were optimized. Information on the internal organization of polytene chromosomes was sought by subjecting isolated preparations to alkali-urea denaturation. Under these conditions, chromosomes gradually extend laterally to give rise to lampbrush-like structures, remarkably similar to the meiotic chromosomes of amphibian oocytes. This effect is at least partially reversed by return of alkali-urea treated preparations to physiological Ringer solution, where chromosomes contract again and near-normal banding patterns are re-established. It is suggested that apart from bulk differences due to polyteny, the basic structure of polytene chromosomes and true lampbrush chromosomes may be rather similar.


The identification of human H-Y antigen and testicular transformation induced by its interaction with the receptor site of bovine fetal ovarian cells. beta 2m(-), HLA (-) Daudi human male Burkitt lymphoma cells excreted a group of protein subunits that shared three distinctive characteristics; their conspicuously longer half-lives compared to more hydrophilic Daudi excreted proteins, their tendency to form progressively larger polymers by means of interchain disulfide bridges, and the extreme hydrophobicity of these polymers. The plasma membrane of extragonadal somatic cells absorbed 1.2 to 2.8% of these hydrophobic proteins. The unoccupied H-Y receptor sites residing on the plasma membrane of bovine fetal ovarian cells, on the other hand, selectively absorbed polymers of 18,000 mol. wt. subunits, and this antigen-receptor interaction, if allowed to continue for five days, induced the formation of tunica albuginea and seminiferous tubules in bovine XX embryonic indifferent gonads. In this manner, human H-Y antigen excreted by Daudi cells has functionally been identified as a series of polymers derived from 18,000 mol. wt. subunits. While, the H-Y antigenic determinants were retained even by the largest polymeric form that became irreversibly water insoluble, the receptor binding activity was shown only by 36.8% of the available polymeric forms of 18,000 mol. wt. subunits, at the most. Nevertheless, once bound to the receptor site, these polymers were rapidly reduced to the monomeric form on the plasma membrane of bovine fetal ovarian cells. Accordingly, the 18,000 mol. wt. monomer might actually represent the functional form of H-Y antigen.


The immature intestine: implications for nutrition of the neonate. The survival and prognosis of the prematurely born human infant are dependent on a successful transition from the intrauterine to the extrauterine environment. This is largely a consequence of the maturation of sufficient gastrointestinal function to provide adequate nutrition. However, the gastrointestinal tract of the premature infant, and to some extent, of the full-term infant, may be unprepared to provide the requisite absorptive function. Data presented in this symposium emphasize the dissociations in the development of human gastrointestinal function. Morphological maturation is completed early in gestation while glucose absorption increases with gestational age. Sucrase and maltase activities appear early; lactase activity begins to increase at 30 weeks and increases steadily to term. The latter pattern is accompanied by increased production of cortisol and thyroid in the fetus. The intraluminal phase of fat digestion is immature even in the full-term neonate. Both pancreatic secretory function and bile salt metabolism mature postnatally. Despite this relative immaturity, breast milk fat is absorbed with great efficiency by the term infant, and breast milk provides other important influences on intestinal development: mitogenic factor, immunological support, control of intestinal flora. The goals of nutrition support of the premature infant have been to maintain intrauterine growth standards; yet premature infants receiving pooled breast milk from mothers at 40 weeks or more may be given too little protein for their needs. Human milk from mothers of premature infants may be a more appropriate nutrient source. Supplements with higher contents of amino acids may lead to amino acid imbalance or hyperammonaemia. Additional stresses and requirements are imposed by illness or congenital anomalies. While we must apply current research findings to clinical care, we must also extend our knowledge of extrauterine human development. The ultimate measure of success in this field will be the physical and neurological capacities of infants followed prospectively.


Cytology and cytochemistry of the hypophysis of the freshwater teleost Rhamdia hilarii (Val., 1840). The characterization of cell types in the rostral pars distalis, proximal pars distalis and pars intermedia of the Bagre fish hypophysis during the reproductive cycles, as well as a cytochemical study of mucosubstances were undertaken. The cytophysiological relationship among the different acidophilic, basophilic and chromophobic cells with the several hypophyseal hormones was also considered.


[Immunopharmacological study of buckwheat hypersensitivity (author's transl)]. Effect of dialysate from buckwheat extract on immediate hypersensitivity reactions. To investigate the hypersensitivity reaction of buckwheat, the aqueous extract of buckwheat was separated into dialysate (BWD) and non-dialysate (BWND). BWD neither decreased a release of anaphylactic mediator from the chopped lung tissue nor inhibited Schultz-dale reaction in the ileum of guinea pig sensitized with BWND. Heterologous PCA mediated by rabbit anti-BWND serum in guinea pig and homologous PCA in rats using anti-BWND IgE serum were inhibited by i.d. or i.v. treatment of BWD. Radioallegosorbent test (RAST) and PK reaction were inhibited in a dose dependent fashion by BWD mixed with human serum sensitive to buckwheat, although BWD required approximately 10,000 times more than BWND to produce a RAST inhibition. RAST, however, was not affected when BWD was added to 125I-labelled anti-human IgE. Anti-buckwheat IgE antibody in sensitized animals and atopic patients was specifically absorbed with an insoluble copolymer of BWD-conjugated BSA. BWD did not form a precipitin line against anti-BWND IgG antibody in immunoelectrophoresis. Rat IgE serum-induced degranulation and histamine release from mast cells were inhibited in a dose dependent fashion by BWD and the inhibition was specific to anti-BWND IgE antibody. BWD may be useful as an hyposensitization agent in buckwheat hypersensitivity, since BWD is a haptenic substance capable of neutralizing specific IgE antibody on mast cells.


[Pharmacological properties of dl-2-(3'-t-butylamino-2'-hydroxypropylthio)-4-(5'-carbamoyl-2'-thienyl) thiazole hydrochloride (S-596), a new beta-adrenergic blocking agent (author's transl)]. We developed a new beta-adrenergic blocking, antiarrhythmic compound S-596 and compared the findings with those of propranolol or practolol. S-596 antagonized the positive chronotropic and inotropic actions of adrenaline in isolated guinea-pig atria, and blocked the relaxant response to adrenaline of isolated guinea pig tracheal strips. In anesthetized mongrel dogs, S-596 given intravenously inhibited increases in heart rate and myocardial contractile force and decreases in systemic blood pressure induced by isoproterenol. In conscious dogs, the oral administration of S-596 reduced the isoproterenol induced tachycardia and the maximal effect was attained one hour after administration. In this regard S-596 was about 5 times more potent than propranolol, and S-596 was significantly longer-acting than propranolol. Thus, S-596 has greater beta-blocking activity than propranolol. S-596 has a lesser degree of myocardial depressant action than propranolol in spontaneously contracting rat or guinea pig atria and no intrinsic sympathomimetic activity in reserpinized rats. S-596 also has a weaker local anesthetic and antiarrhythmic activity than propranolol, as determined in guinea pigs and rabbits.


Immunodiffusion systematics of the primates. Part V. The Platyrrhini. Evolutionary relationships between New World monkeys and marmoset genera and the place of the Ceboidea within the primates are considered in terms of the immunological specificity of ceboid proteins. Antigenic distances between the New World primates are measured using antisera produced in rabbits to nine ceboid genera: Alouatta, Aotes, Ateles, Callicebus, Cebus, Chiropotes, Lagothrix, Saimiri and Saguinus. A cladogram constructed on the basis of increasing degrees of antigenic distance between branches depicts Ceboidea as a monophyletic assemblage within which Alouatta is grouped with the Atelinae genera, Lagothrix and Ateles, Chiropotes joins Cacajao and Cebus joins Saimiri. The joining of the cebid genera Aotes and Callicebus with callithricid genera Callimico and Saguinus into a single complex lineage suggests that Cebidae is a paraphyletic or polyphyletic taxon. A phylogenetic taxonomy for Platyrrhini is proposed.


[Effect of kidney function on the elimination and action of colloidal plasma substitutes]. Hypovolemia and prophylaxis of thrombosis are the main indications for colloidal plasma substitutes. This is also true for patients with renal failure. Due to their difference in molecular structure colloidal plasma substitutes, which are in clinical use--dextran, hydroxyethyl starch, gelatin--, are eliminated tbrough different pathways. However, the kidney plays an important role in the elimination of all three substances. Therefore pharmacokinetics and pharmacodynamics of colloidal plasma substitutes, which are based on dextran, hydroxyethyl starch and gelatin, were studied in patients with normal or impaired renal function. Renal insufficiency proved to affect the elimination and therapeutic efficacy of these colloids in different and characteristic ways. Consequently this requires special indications and dosage recommendations for patients with renal insufficiency.


Induction of antibody responses in vivo by antigen specific helper factor. T helper cell factors (HF) have been preparated against protein and synthetic antigens by restimulating in vitro induced helper cells with small amounts of antigen. HF when tested in vitro are antigen specific and capable of replacing T cells, initiating a thymus dependent IgM response. In the in vivo adoptive transfer assay HF is capable of replacing T helper cells and is active at very high dilution, inducing both IgM and IgG responses. When tested on unirradiated DNP primed animals the HF was able to initiate a thymus dependent anti-hapten response, comparable to that seen with helper T cells, of both IgM and IgG class. It was also shown that HF acts on anti-Thy 1 treated spleen cells. The results indicate that antigen specific helper factors may be one of the physiologic mediators of T-B interactions in intact animals.


Hypothesis on the origin of the strong alloreactivity. The high proportion of alloreactive T lymphocytes and many of the available data on T cell receptors can be explained by one single hypothesis with four basic assumptions: A) The functional induction of T lineage cells in the thymus inherently causes a selection for V-regions that bind to major histocompatibility antigens (MHA). The type of MHA determines the functional pathway of the T cell. B) This process selects with the highest probability for binding sites with high affinity for the self-MHA, yet binding sites with high affinity for non-self-MHA and low affinity for self-MHA will also be selected with a low but finite probability. C) This positive selection for self-MHA binding V-regions is followed by a rigorous selection against self-reactive T cells during the subsequent thymic or post-thymic phase of tolerance induction. D) Most crucial for the hypothesis is, finally, the assumption that the second (negative) selection operates with a higher affinity threshold than the first (positive) selection. The negative selection thus spares T cell clones with low affinity for self-antigens. This provides a strong selective advantage for two major groups of cells, namely alloreactive cells most of which recognize nonself-MHA in complex with nonpolymorphic non-MHA determinants, and cells that recognize nonself-determinants in complex with self-MHA with different degrees of restriction. One of the predictions of this hypothesis is that the proportion of alloreactive cells is relatively small among the T lineage cells that leave the thymus but increases largely during the post-thymic development of the peripheral T cell pool. The hypothesis is not biased in respect to the underlying germ line repertoire of V genes, and is in fact compatible with the simple assumption that T and B cells use the same sets of V genes.


Studies of the producer cell of interferon in human lymphocyte cultures. The producer cells of interferon were studied in human leucocyte cultures stimulated by a variety of stimulants, including phytohemagglutinin (PHA), pokeweed mitogen (PWM), Corynebacterium parvum (CP) and Herpes Simplex Virus (HSV). When the cells were fractionated by the use of neuraminidase-treated sheep red blood cells (SRBC), the T cell population responded with interferon production to PHA and PWM but not to CP or HSV. However, the non-T population showed a vigorous response to the latter two stimuli. In contrast, nylon column eluate cells enriched for T cells responded well to CP and HSV with production of interferon. To resolve these contradictory data, we have used combinations of techniques. Nylon column eluate cells were further separated by SRBC and it was found that the nylon non-adherent rosetting cell did not produce interferon in response to HSV or CP whereas the nylon-nonadherent non-rosetting cell did. In additional experiments more elaborate techniques were used. Leucocytes were treated by plastic adherence and iron filings, passed over a nylon column and subsequently over an Ig-anti-Ig column, and then rosetted with SRBC. Again only the non-rosetting population produced interferon. In parallel experiments the capacity of the different cell populations to lyse three types of target cells in a chromium release assay as a test for "natural killer" (NK) cell activity was investigated. There was some correlation between interferon production and NK cell activity. Thus, our data indicate that interferon is produced by non-T, non-B cells, possibly by cells related to NK cells.


Studies on renin activation in normal human plasma. The phenomenon of plasma renin activattion by acid dialysis and preincubation with trypsin was studied in normal human plasma. Activation of plasma renin by exposure to pH 3.3 was shown to require at least one dialysis step and could be inhibited by the presence of Trasylol, indicating the involvement of a protease in acid activation. Amniotic fluid exposed to pH 1.5 to destroy renin and renin substrate was also found to contain an enzyme capable of activating plasma renin. The Michaelis-Menten constant Km and the molecular weight of activated "renin" were found to be similar to those of normal plasma renin. Inactive renins or renin-like enzymes were partially purified from plasma by affinity chromatography on concanavalin A, precipitation with (NH4)2SO4 and isoelectric focusing. Trypsin and acid exposure gave similar results with regard to the activation of this zymogen, suggesting that trypsin and acid dialysis may increase plasma renin activity by the same mechanism.


[Improvement of blood viscosity after infusion of low-molecular hydroxyethyl starch (Expafusin) in healthy subjects]. In healthy volunteers, 500 ml of a 6% low molecular weight hydroxyethyl starch solution (Expafusin) was infused intravenously within 30 minutes. Blood samples for the measurement of rheological parameters were obtained before and after the infusion. Blood and plasma viscosity were significantly decreased after the infusion of HES. Red cell deformability as measured by a filtration technique was slightly improved. Red cell aggregation as measured by a light transmission method was markedly reduced. Repeated infusion of 500 ml Expafusin indicates that the improvement of the flow properties of blood can be maintained for more than 3 hours.


[Effect of various plasma substitutes on blood groups]. In order to test the influence of the plasma substitutes Macrodex 6%, Plasmasteril and Expafusin on subsequent blood-typing and serologic tolerance tests, blood specimens of known donors of blood group 0 Rh negative were incubated with the respective plasma substitutes in various dilutions and serologic tests performed. In the AB0 and Rh system, none of the plasma substitutes tested caused falsely positive agglutinations. Reading of cross-match in the albumin test was not impeded in any of the substitutes. The higher molecular dextran Macrodex 6% and the high molecular hydroxyethylene starch Plasmateril appeared to have positive blood-group serologic reactions. With the low molecular hydroxyethylene starch Expafusin no irritations occurred in all tests of cross-matching.


Rapid dextran infusion in essential hypertension. Hemodynamic parameters were studied before and after rapid dextran infusion in 34 men including 17 patients with sustained essential hypertension and 17 normotensive controls. In both groups of patients, dextran infusion induced a significant increase (p less than 0.001) in central venous pressure (CVP), cardiac output (CO), and stroke volume. The percent change in stroke volume was significantly higher in hypertensives (p less than 0.001) than in controls. Three indices of volume expansion were calculated: 1) the ratio between the change in CO and the change in volume, which was significantly higher in hypertensives (p less than 0.025), 2) the ratio between the change in CO and the change in CVP, which was similar in both groups, and 3) the ratio between the change in volume and the change in CVP, which was significantly reduced in hypertensives (p less than 0.001). In the overall population, the latter ratio was negatively correlated with the change in CO (or in stroke volume) induced by expansion ( r = -0.75). The results provided evidence that: 1) the slope of the relationship between CO and blood volume was steeper in hypertensives than in normotensives, and 2) the steeper slope was due to a reduction in the effective compliance of the vascular bed, causing a greater elevation in CO per unit rise in volume.


Six years' experience with cryosurgery in the oral cavity. Ninety selected patients with a benign or malignant lesion in the oral cavity were treated by cryosurgery. A clinical study was carried out during a 6-year period to investigate the value of cryosurgery as supplemental or substitute therapy. Good results have been obtained in the treatment of small to moderate, superficially situated angiomas. Cryotherapy has also been found to be satisfactory in the treatment of papillary hyperplasia of the palate. As symptomatic treatment we employed the freezing procedure to painful erosive lichen planus. Until now, a casual treatment has not been possible because of an unknown etiology. Used as symtomatic therapy, cryosurgery may be of some use in these cases, especially to relieve pain. Good results have been obtained in the treatment of oral leukoplakia. The pathologically changed mucous membranes could be completely eliminated in most of the cases without severe scar formation or impairment to functions. Cryosurgery for palliation was employed in 11 incurable tumors. Results have been disappointing. Twenty-one localized malignant neoplasms were treated by cryosurgery to cure. The tumor was completely destroyed in 67% of the cases.


The surgical treatment of cancer of the esophagus. Summary of replies to a questionnaire. In an attempt to assess the different methods of treatment of tumors of the esophagus we contacted 170 surgeons (in 1975-1976). Seventy-six (45%) answered a questionnaire concerning the surgical management of esophageal tumors, the reasons for non operability and non resectability, the place of radiotherapy, the validity of palliative procedures, the exposure and type of resection for tumors at various levels in the esophagus, the technical details of the operations, the resectability and mortality rate, the anastomotic leak rate and its treatment, and the five-year survival rate after operation. The replies to the questionnaire are summarized.


1-N-Alkyl analogs of butirosin.

1-N-Alkyl analogs of butirosin. Butirosin (Ia), 5''-amino-5''-deoxy-(Ic), 3',4'-dideoxy-(Ie), and 5''-amino-3',4',5''-trideoxy-butirosin A (If) were converted into the corresponding 1'''-deoxo derivatives, Ib, Id, Ig, and Ih by borane reduction. In addition, xylostasin (IIIa) was converted into its 1-N-ethyl derivative (IIIb) by reductive ethylation. Their antibacterial activities were discussed.


The biochemical effect of auromomycin on bacterial cells. The mechanism of action of auromomycin was studied with intact cells of Bacillus subtilis. The antibiotic exhibited a preferential inhibition of DNA synthesis over RNA and protein syntheses. The strand scission of cellular DNA was induced by the drug, and DNA was degraded into acid-soluble fragments. The results suggested that DNA is the chemoreceptor of auromomycin.


The distribution of desmosomes and ruthenium red-bound cell surface carbohydrates during palatal closure in the hamster. The distribution of ruthenium red (RR)-bound cell surface carbohydrates and desmosomes was studied during epithelial adhesion and seam formation stages of palatal development in the hamster. Before closure, RR-positive material was observed on the plasma membrane of the superficial cells and in the intercellular spaces of the palatal epithelium. At the time of initial contact, RR-bound substance was absent or reduced on the outer plasma membrane and desmosomes were forming between the epithelial cells of the opposing palatal shelves. Except in the oral and nasal triangular areas, RR-positive material was absent in the epithelial seam. Quantitative analysis showed a three-fold increase in the number of desmosomes from the vertical stage to the epithelial fusion stage which correlated well with the disappearance of RR-bound material. Desmosome formation appears crucial during initial adhesion between epithelia of the opposing palatal shelves and during the formation and maintenance of the seam.


The cyclic AMP-dependent initiation of DNA synthesis by T51B rat liver epithelioid cells. The brief rise in the cellular cyclic AMP content which occurs late in the prereplicative phases of rat hepatocytes in vivo and T51B rat liver epitheloid cells in vitro seems to be necessary for the initiation of DNA synthesis. Thus, the extracellular calcium-deprivation in T51B rat liver cells in culture which induces a late G-1 block is rapidly reversible (cells surge into S phase within one hour) either by creating a cyclic AMP surge by the addition of calcium or 3-isobutyl-1-methyl xanthine (a cyclic 3',5'-nucleotide phosphodiesterase inhibitor) or by the exogenous addition of low concentrations of cyclic AMP itself (i.e., 10(-8)-10(-5) M). On the other hand, prevention of the calcium-induced cyclic AMP surge by imidazole (a cyclic 3',5'-nucleotide phosphodiesterase activator) blocked the initiation of DNA synthesis by the calcium-deprived T51B cells.


Role of hydrogen bonding in red cell aggregation. The role of hydrogen bonding in red cell aggregation induced by dextran was studied with the use of urea, an inhibitor for hydrogen bonding. In order to avoid hemolysis of red cells by the high concentration of urea, the studies were performed on human red cells hardened in glutaraldehyde. The degree of red cell aggregation at Hct = 45% was estimated by the use of a coaxial cylinder viscometer. The viscometric aggregation index (VAI) was calculated from viscosity values at shear rates of 52 sec-1 (eta H) and 0.05 sec-1 (eta L); VAI = (eta L - eta H)/eta H. Red cells with surface charge intact and with charge removal by neuraminidase treatment were studied. Urea at high concentrations, e.g., 6 M, significantly inhibited red cell aggregation induced by dextran. These findings indicate that hydrogen bonding plays an important role in dextran-induced red cell aggregation. An understanding of the nature of the forces involved in red cell aggregation serves to establish the physicochemical principles of cell-to-cell interactions induced by macromolecules.


Electrochemical detection of tryptophan metabolites following high-performance liquid chromatography. Electrochemical detection of tryptophan metabolites following separation on a reversed-phase high-performance liquid chromatography column was compared with other means of detection. Of 29 compounds studied, 26 could be detected at a sensitivity comparable to that of fluorescence derivatisation procedures. Response was linear over a wide range of concentrations and selectivity was shown to be superior to ultraviolet detection when analysing urine. Additionally, it was possible to control selectivity so that only those tryptophan metabolites from the tryptophan hydroxylase pathway were detected. This is of particular value in the study of distrubances of serotonin metabolism and is unique to this type of detector.


Antigenic components of Taenia saginata and their relevance to the diagnosis of bovine cysticercosis by immunoelectrophoresis. 22 antigenic components of Taenia saginata proglottides were detected by immunoelectrophoretic (IEP) study. Cross-absorption of the hyperimmune serum, raised in rabbits against T. saginata, with heterologous and host antigens showed that two thirds of these components were not specific for the cestode. Among the calves experimentally infected with T. saginata eggs, 3 precipitation patterns in IEP were identified depending on the evolutionary stage of the infection. The hydrosoluble extract of T. saginata was used as antigen. A short precipitating arc present near the antigen well and appearing 3 to 4 weeks post-infection of the calves and a long precipitating arc extending towards the anodic end of the slide and appearing in later stages of infection were found of high diagnostic value. No false-positive reaction or cross-reaction were found. Among the calves harbouring less than 50 cysticerci, false-negative reactions were seen.


Immune response to herpesvirus antigens in adults with acute cytomegaloviral mononucleosis. The immune response of eight patients with mononucleosis caused by cytomegalovirus (CMV) was measured early in their illness--when virus was present in their urine and/or blood--and subsequently during convalescence. Levels of CMV-specific antibody rose early in the illness, but the proliferative response of mononuclear cells to CMV antigen did not reach the level characteristic of CMV-immune donors until several months later. The production of interferon by mononuclear cells in response to CMV antigen was also low early in the illness. Although these patients had prior immunity to herpes simplex virus and varicella-zoster virus, their mononuclear cells responded poorly to antigens prepared from these viruses. The proliferative response to these antigens returned to normal in parallel with the development of a normal response to CMV. It is suggested that acute CMV mononucleosis suppresses the proliferative response of human mononuclear cells.


[The antenatal diagnosis before 20 weeks of an omphalocoele and a failure of development of the abdominal wall (author's transl)]. We report 2 cases of malformations occuring in the region of the umbilicus which were diagnosed by amniocenteses before the 20th week of pregnancy (with levels of alphafetoprotein) and echography. Failure of development of the abdominal wall was only diagnosed for certain after the therapeutic abortion had been carried out. The omphalocoele was diagnosed for certain by echography at the 19th week after anencephaly and spina bifida had been eliminated in spite of an abnormally high rise in the level of amniotic fluid alphafetoprotein (100.000 microgram/l). We have not found any similar cases in the literature at this gestational age. These observations prove the value of combining the estimation of alphafetoprotein level in the amniotic fluid when making antenatal diagnoses because of the age of the mother (after the age of 38), with echography specially directed to searching for such apparent morphological abnormalities.


Effect of specific antibodies on chronic echovirus type 5 encephalitis in a patient with hypogammaglobulinemia. The effects of specific antibodies in chronic echovirus type 5 (echo 5) encephalitis were investigated in a patient with x-linked hypogammaglobulinemia. Virus was detected in cerebrospinal fluid (CSF) and blood despite treatment with commercial human gammaglobulin that contained low titers of antibodies to echo 5 (0.6 x 10(4) units per injection). Virus disappeared from blood and CSF when plasma containing high concentrations of antibodies (total dose, 1--4 x 10(4) units/kg) was administered intravenously. Maximal inhibition of virus was achieved in culture and in the patient's CSF when the titer of antibody to echo 5 in CSF was greater than or equal to 16 units/ml. Although the patient died, hyperimmune plasma improved the neurologic status and eliminated detectable virus from the blood and CSF.


Chemotherapy of herpes simplex virus encephalitis in mice by direct intracerebral injection. The effects of 9-beta-D-arabinofuranosyladenine-5'-phosphate (ara-A-5'-P) on herpes simplex virus encephalitis in mice were studied. This drug is a monophosphate derivative of adenine arabinoside (ara-A). When administered by the intracerebral (ic) route, ara-A-5'-P was highly effective in virus-infected mice in the 17-20 g weight range provided therapy was begun before clinical symptoms of encephalitis appeared. The most effective dose schedule was found to be three daily ic injections totaling 400 micrograms/day (20 mg/kg) beginning on the third day after injection. Such treatment resulted in a survival rate of 50%-75% among infected mice after six months. In the early stages of many experiments, 90%-100% of the controls died whereas all of the treated animals survived.


Chlamydia psittaci: growth characteristics and enumeration of serotypes 1 and 2 in cultured cells. The growth characteristics of Chlamydia psittaci serotypes 1 and 2 (ovine and bovine origin) were studed in mouse L cells. Formation of inclusions and yield of infectious progeny for serotype 1 were maximal when host cells were treated with cycloheximide and the pH in the cell culture medium was 7.2-7.4. The number of cells that contained inclusions and the infectivity yield for serotype 2 were maximal when the pH was 6.6-7.0. Treatments with diethylaminoethyl dextran and cycloheximide increased inclusion formation in serotype 2 but decreased the yield per infected cell. For both serotypes, centrifugation of the chlamydiae onto monolayers infected 50-1,200 times the number of cells infected with use of stationary adsorption. An enumeration method for L-cell infectious units was developed that gave higher titers than the chicken embryo 50% lethal dose after yolk sac inoculation.


Simultaneous occurrence in the same serum of hepatitis B surface antigen and antibody to hepatitis B surface antigen of different subtypes. The simultaneous occurrence of hepatitis B surface antigen (HBsAg) and antibody to HBsAg (anti-HBs) of different subtypes in the serum of a hemodiaylzed patient was studied. The w(a) subdeterminants were involved. The HBsAg belonged to the ayw3 subtype, and the anti-HBs exhibited monospecific anti-w2 activity. Both the HBsAg and the anti-HBs were detectable by counterelectrophoresis (CEP). The specificity of the antibody was demonstrated by CEP in tests against 128 sera containing HBsAg of 12 different subtypes and in absorption experiments with eight sera containing HBsAg of eight different subtypes, as well as by radioimmunoassay in the liquid phase. The monospecific antibody was selectively directed against the w2 subdeterminant of the adw2 subtype and was designated anti-w2.


Immunodiagnosis of systemic candidiasis: mannan antigenemia detected by radioimmunoassay in experimental and human infections. A radioimmunoassay (RIA) that detects candida mannan was developed so that immunodiagnosis of systemic candidiasis could be improved. The RIA was evaluated in an animal model of disseminated disease and in a panel of patient sera. Mannan antigenemia was detected with the RIA in 52% of 29 rabbits with systemic candidasis, but not in 60 normal rabbits or 31 rabbits with systemic aspergillosis. In an evaluation of human sera, mannan antigenemia was detected in five of 11 patients with systemic candidiasis, one of three patients with invasive gastrointestinal candidiasis, and one patient with a sustained candidemia associated with an infected intravenous catheter. Mannan was not detected in sera from 11 patients with superficial candida infections, seven patients colonized with Candida, three patients with chronic mucocutaneous candidiasis, eight patients with other systemic mycoses, or 22 normal donors. This study demonstrates the utility of this RIA for early, specific immunodiagnosis of invasive candidiasis.


Relationships among raffinose plasmids determined by the immunochemical cross-reaction of their alpha-galactosidases. Plasmid-encoded alpha-galactosidase served as a marker enzyme for the recognition and comparison of raffinose (Raf) plasmids present in strains of Escherichia coli. Immunochemical relationships were established among Raf plasmids of 39 independent isolates from man and domestic animals (from three continents) by using antiserum against alpha-galactosidase. Immunodiffusion revealed three serological subclasses of alpha-galactosidase, which are correlated with the biological and geographical origin of the host strains. It is concluded that the raf determinants of all Raf plasmids tested have evolved from a common ancestor.


Phosphonoformate inhibits reverse transcriptase. The new antiviral substance phosphonoformate (PFA) has been tested in a cell-free system for its effect on reverse transcriptases from an avian retrovirus (avian myeloblastosis virus, AMV) and from mammalian retroviruses (Rauscher leukaemia virus, RMuLV; bovine leukaemia virus; baboon endogenous virus; simian sarcoma virus; visna virus). The observed inhibitory effect of PFA has been compared with that of a structurally related substance, phosphonoacetate (PAA). Phosphonoformate, at a concentration of 100 microM, reduced the activities of all the above mentioned polymerases by 90% when (rA)n.(dT)10 was used as a template/primer. The dose-response curves for AMV and RMuLV polymerases primed with (rA)n.(dT)10 showed PFA to be a 1000-fold more active than PAA; the RMuLV polymerase activity was reduced to 50% after incubation with 0.7 microM-PFA and 0.7 mM-PAA, respectively. There was no difference in PFA inhibition of virus-associated and purified reverse transcriptase activity. Results with various synthetic templates showed that both the RNA- and the DNA-dependent polymerase activities of reverse transcriptase were inhibited by PFA. The endogenous polymerase activity of AMV was inhibited to 50% at 100 microM-PFA, while PAA had no effect. The PFA inhibition was dependent on whether Mg2+ or Mn2+ was used as divalent cation in the assay. Phosphonoformate arrested DNA synthesis immediately after being added to the assay system. The mechanism of inhibition of the AMV polymerase was non-competitive with respect to substrate and template and the apparent inhibition constants were 16 microM and 9 microM, respectively.


Priming increases the amount of interferon mRNA in poly(rI).poly(rC)-treated L cells. Priming by mouse interferon pre-treatment resulted in an accumulation of interferon mRNA in poly(rI).poly(rC)-treated L cells, starting early in the period of interferon synthesis. On electrophoresis, the priming activity of an interferon preparation co-migrated with the antiviral activity, which suggests identity of the functional principle(s) for these activities.


Somatic cell genetics of human interferon production in human-rodent cell hybrids. Forty-two primary human-mouse cell hybrids, derived in two separate experiments, were treated with Newcastle disease virus (NDV): eight hybrids were found to produce human interferon and this was shown in every case to be predominantly of the fibroblast type. An extensive analysis was made in terms of karyotype and marker enzymes on all the eight hybrids producing interferon and also on five hybrids which did not produce interferon, five randomly selected hybrids and eleven subclones resistant to diphtheria toxin. The results suggest that, contrary to previous reports, a gene on chromosome 5 is not involved in production of human interferon. Its production was however correlated with the presence of chromosome 9 in the hybrids. Analyses of two sets of human-Chinese hamster hybrid subclones from two different crosses were also consistent with the assignment of a human interferon gene to chromosome 9.


Modulation by cell trypsinization of Sendai virus expression in African green monkey kidney cells: first infection and establishment of a carrier state. Two African green monkey kidney (AGMK) cell lines, 37RC (interferon-producing) and Vero (non-interferon-producing), were infected by egg-grown Sendai virus passaged in eggs at high and low m.o.i. The appearance of haem-adsorption, and cytopathic effect (c.p.e.) as well as the presence of haemagglutinating virions in the supernates were much more pronounced with a virus seed obtained with 10(-3) diluted passages than with a seed obtained with undiluted inoculum. They were also independent of interferon production (data obtained in 37RC and Vero cells were almost superimposable). In studies carried out with the virus seed prepared at high dilution the establishment of infection was maximal when monolayers were infected as soon as 5 h after trypsinization and cell seeding, regardless of cell density. Virus in supernates obtained from cultures infected 5, 20 or 50 h after seeding exhibited a greatly reduced infectivity for monkey cells, but not apparently for chick embryos. Trypsin treatment of the virus supernates restored their infectivity for AGMK cells more efficiently for virus released from cells infected 5 h after seeding than for virus released from cells infected later after seeding. In keeping with these observations, virus in supernates from cultures infected 5, 20 or 50 h after seeding contained increasing amounts of auto-interfering virions. The decreased infectivity obtained in cell supernates was not accounted for by major differences in virus RNA synthesis. Similarly, the optimum infection established in cells seeded only for 5 h was not due to increased virus adsorption. Several lines of cells surviving first infection with egg-grown Sendai virus have been obtained and kept in cultures for 3 to 18 months. Virus release and c.p.e. apparently become reduced in the cells surviving the first infection until the newly repopulated monolayers must undergo trypsinization. Weekly protease treatments of the cells reactivate all parameters of virus infection which again will tend to disappear slowly and then reappear following each trypsinization ('intermittent' carrier state). The establishment and the 'intermittent' reactivation of these lines were not prevented by the inclusion in the medium of anti-Sendai virus serum. Temperature-sensitive ts functions do not seem to play an important role in this virus-host relationship.


Comparative biochemical and serological analysis of five isolates of a single serotype of foot-and-mouth disease virus. A comparison has been made of some of the biochemical and serological characteristics of five isolates of foot-and-mouth disease virus (FMDV), serotype A. Three of the viruses have been assigned to the same subtype, A22; the other two belong to different subtypes, A5 and A24. RNA competition hybridization and two-dimensional electrophoresis of the oligonucleotides produced by ribonuclease T1 showed that the three A22 viruses formed a group which could be distinguished from the A5 and A24 viruses. However, the three A22 viruses showed some differences by both tests. Analysis of the virus polypeptides by polyacrylamide gel electrophoresis methods also distinguished the A22 viruses as a group distinct from the A5 and A24 viruses, but small differences within the A22 group were observed using electrofocusing techniques. Serological differences were observed between the viruses using complement fixation tests and by competition radioimmunoassay with antisera obtained from guinea pigs infected with these viruses. The greatest similarity occurred between the viruses previously subtyped as A22, with A5 and A24 being distinct from the A22 group and from each other. The relationship of the biochemical and serological data is discussed.


Type-C virus antigen detection in co-cultures of human leukaemic bone marrow and dog cells. Bone marrow of leukaemic patients, non-leukaemic patients and normal individuals were co-cultivated with the canine cell line A7573. These co-cultures were screened for retrovirus antigens by means of the indirect cytoplasmic immunofluorescence assay (IFA). Rabbit antisera directed against the major structural protein (p30) of woolly monkey (simian) sarcoma leukaemia virus (grown in human lymphoid cells) and Rauscher murine leukaemia virus were used for testing. After 2 months in culture, 6 of 17 co-cultures containing cells from leukaemic patients showed positive staining in the IFA with the anti-simian virus serum. In control dog cells fluorescence was never observed. Five of the six positive cultures were derived from leukaemic children. One of 12 co-cultures of the non-leukaemic group and one of nine normal bone marrow co-cultures were positive with the simian virus antiserum. None of the 38 co-cultures stained positive in the IFA with Rauscher virus antiserum. Absorption of the simian virus antiserum with calf serum or mouse mammary tumour virus had no dramatic effect in the IFA on positive control cells or on cells of a positive co-culture. However, absorption with purified simian virus (grown in rat cells) completely abolished these fluorescence reactions. The results provide evidence that simian sarcoma-leukaemia virus related information was present in the original bone marrow samples and that co-cultivation with permissive mammalian cells enabled the detection of virus footprints.


Biochemical properties of paramyxovirus Duck/Mississippi/75 neuraminidase. The neuraminidase activity of two strains of Duck/Mississippi/75 virus:DK/Mississippi/320 and DK/Mississippi/334 was studied. These neuraminidases hydrolyse the alpha 2 leads to 3 and alpha 2 leads to 8 ketosidic bonds of different substrates such as fetuin, N-acetyl neuramine lactose and colominic acid, but do not hydrolyse the alpha 2 leads to 6 bonds of mucin type I and type II. The kinetic values of the neuraminidases, Michaelis constant, maximal and initial velocities and the effect of pH, temperature and detergents were also evaluated. The isolates differ mainly in the optimal pH and temperature conditions of activity. As with other paramyxovirus neuraminidases, the enzyme of DK/Mississippi/75 was destroyed by ionic but not non-ionic detergents.


Comparative studies on LS antigens induced by vaccinia and cowpox viruses. The constituents of LS antigen from cells infected with vaccinia virus and with cowpox virus were compared by immunoprecipitation and SDS-polyacrylamide gel electrophoresis (SDS-PAGE). Antiserum to the LS antigen from cells infected with vaccinia virus reacted with at least five polypeptides in cells infected with either virus. Four of these polypeptides were similar sizes in cells infected with the two viruses. However, one major polypeptide with a mol. wt. of about 100 000 (100 K) detected in cells infected with vaccinia virus was not found in cells infected with cowpox virus. Conversely, a polypeptide with a mol. wt. of about 60 000 (60K) was detected only in cells infected with cowpox virus.


Comparison of the antibodies elicited by the individual structural polypeptides of foot-and mouth disease and polio viruses. Antibody produced against preparations of VP1, one of the four structural polypeptides of foot-and-mouth disease virus, neutralized the virus and reacted with both full and empty particles in radioimmunoassays (RIA). Antiserum against VP2 reacted with artificial empty particles of the virus but not with full particles. In contrast, none of the individual polypeptides of poliovirus produced antisera which neutralized the virus nor reacted with it in RIA. However, antisera produced with VP1 and VP2 reacted with artificial empty particles in RIA.


Purification of adenovirus 4 type-specific hemagglutinins for use in diagnostic counterelectrophoresis tests. Three type-specific hemagglutinins of adenovirus (AV) 4 were purified, tested for various activities, and used as immunogens in rabbits. The components were purified by anion exchange and exclusion chromatographies to purification factors of 33, 20, and 37, respectively, and were identified as fiber (polypeptide, mol wt 70,000; incomplete hemagglutinin), penton (mol wt greater than 400,000; incomplete hemagglutinin; possession of cytotoxic activity), and dodecon (mol wt greater than 400,000; complete hemagglutinin; appearance by electron microscopy). Rabbit antisera to these hemagglutinating components possessed type-specific hemagglutination-inhibition and serum neutralization antibodies and were thoroughly evaluated for potency and specificty in immunodiffusion (ID), immunoelectrophoresis, and counterelectrophoresis (CE) tests. The anti-fiber and anti-dodecon sera were highly type-specific in ID and CE tests with a battery of commonly isolated AV serotypes; the anti-penton serum was similarly specific after it was absorbed with AV 2 hexons. These sera were thus proved to be suitable reagents for use in both the routine diagnostic ID test and the more rapid CE test.


Uptake of thallium-201 in enlarged thyroid glands: concise communication. We have investigated the thyroid uptake of Tl-201 in 37 patients with various types of goiter, and in six with normal thyroids. Significant thallium uptake was found in all cases in which there was thyroid enlargement, including Graves' disease, toxic thyroid nodule, primary hypothyroidism, simple goiter, Hashimoto's disease, thyroid carcinoma, and thyroid adenoma. If goiter was absent, however, there was no demonstrable uptake--e.g., in secondary hypothyroidism, subacute thyroiditis, and the normal controls. Thallium uptake did not correlate with thyroid function tests such as BMR, T3-RU, T3, T4, TSH, antithyroid antibodies, or the 24-hr I-131 uptake. In 23 patients with diffuse goiter, on the other hand, maximum Tl-201 uptake correlated well with thyroid weight: r = 0.836 (p less than 0.001); y = 0.02 x + 0.06.


Mice with spontaneous mammary tumors develop type-specific neutralizing and cytotoxic antibodies against the mouse mammary tumor virus envelope protein gp52. Sera from C3H mammary tumor-bearing mice contain cytotoxic antibodies for mouse mammary tumor virus (MMTV)-producing cells, based on (51)Cr release in a complement-dependent serum cytotoxicity assay. The cytotoxic antibodies could be absorbed by purified C3H MMTV gp52 and C3H MMTV-infected cat cells (C3H [MMTV] CrFK) containing cell surface MMTV gp52. However, purified MMTV p27 and uninfected CrFK cat cells were negative. Absorption of the sera with GR (MMTV) CrFK cells also removed all of the cytotoxicity, whereas absorption with RIII (MMTV) CrFK cells was negative, even though all three infected cat cells contained equivalent amounts of gp52. The same C3H cytotoxic sera also neutralized the focus-forming capacity of a C3H MMTV pseudotype of Kirsten sarcoma virus containing MMTV gp52. In contrast, sera from mammary tumor-bearing GR and RIII mice did not neutralize the pseudotype. Furthermore, neutralization could be achieved only by anti-gp52 but not by anti-gp36, -p27, -p14, or -p10 C3H MMTV sera. The gp52's of C3H, GR, and RIII MMTV could also be distinguished by using a type-specific competition radioimmunoassay employing (125)I-gp52 of C3H MMTV and a hyperimmune rabbit anti-C3H MMTV serum. To demonstrate these differences directly, we studied the primary structure of gp52 on the surface of the C3H, GR, and RIII (MMTV) CrFK cells. Two-dimensional tryptic peptide maps of the cell surface lactoper-oxidase-catalyzed iodinated gp52's revealed a greater number of peptides common to the gp52's of C3H and GR MMTVs than to RIII MMTV gp52. These results demonstrate that gp52 is a major target antigen for both cytotoxic and neutralizing antibodies, that the cell surface and virion-associated gp52's of C3H, GR, and RIII MMTV contain both group- and type-specific determinants, and that C3H and GR MMTV gp52's are antigenically more related to each other than to RIII MMTV gp52. Furthermore, C3H mammary tumor-bearing mice develop type-specific antibodies capable of recognizing unique gp52 determinants and, therefore, are able to distinguish the gp52 of C3H MMTV from the gp52's of GR and RIII MMTV.


Characterization of 40,000- and 25,000-dalton intermediate precursors to Rauscher murine leukemia virus gag gene products. Under steady-state labeling conditions, Rauscher murine leukemia virus-infected NIH Swiss mouse cells contain at least three major polyproteins derived from the viral gag gene. They have molecular weights of 65,000, 40,000, and 25,000. They have been termed pPr65gag, Pr40gag, and pPr25gag. pPr65gag has been shown by a number of laboratories to be composed of all four core proteins (p15, pp12, p30, and p10). In this paper, Pr40gag was found to contain p30 and p10 antigenic determinants and peptide sequences, whereas pPr25gag was found to contain p15 and pp12. Pr40gag and pPr25gag are rapidly labeled precursor proteins that were detectable early in pulse-chase experiments. Both precursors disappeared during the later stages of the chase period concurrent with the appearance of the mature viral core proteins. pPr65gag and pPr25gag were found to be phosphorylated, pPr25 having a higher specific activity of 32P than pPr65. In spite of this, peptide mapping studies, as well as the identification of the phosphorylated amino acid residues of pPr65, and pPr25, and pp12, indicated that the same sites are phosphorylated regardless of whether the precursors or the mature pp12 are examined.


Hepatitis B viral DNA molecules have cohesive ends. Hepatitis B virus DNA made fully double stranded by a virion DNA polymerase reaction could be converted from circular to linear molecules by heating in 10 mM NaCl at 77 degrees C or in 100 mM NaCl at 90 degrees C for 15 min. Heat-generated linear hepatitis B virus DNA was reannealed to circular molecules by incubating in higher salt concentrations. The identity of the molecular forms was established by their electrophoretic mobility and appearance in electron micrographs. Recircularization was blocked by reacting linear molecules with nuclease S1 or avian myeloblastosis virus reverse transcriptase. These results suggest that the heated linear DNA had single-stranded ends with complementary nucleotide sequences. It also suggests that a discontinuity or nick is present in each strand of the circular DNA molecule after the single-stranded region is made double stranded by the virion DNA polymerase reaction. The difference in contour length by electron microscopy of circular and linear molecules spread under aqueous conditions suggested that the discontinuities in the two strands were about 270 base pairs apart. The amount of nucleotide incorporated into the ends of heat-generated linear hepatitis B virus DNA by reverse transcriptase suggested that the single-stranded ends were about 305 bases in length. This fully double-stranded linear DNA was cleaved with EcoRI or HpaI restriction endonuclease. The sum of the two fragments generated by each totaled 3,510 base pairs, 310 base pairs greater than the contour length of circular hepatitis B virus DNA which represents a third estimate of the distance between the discontinuities in the two DNA strands of circular DNA. Restriction endonuclease cleavage also indicated that the ends of heated linear DNA which correspond to the discontinuities in the two strands of the circular DNA are at unique sites in the DNA with respect to the restriction sites.


Serological relationship of woodchuck hepatitis virus to human hepatitis B virus. Two antigenic systems of the woodchuck hepatitis virus have been identified. The relationship between viral antigens of the woodchuck hepatitis virus and the human hepatitis B virus was determined by using immunoprecipitation, hemagglutination, and immune electron microscopy techniques. Antigens found on the cores of the two viruses were cross-reactive. Lack of cross-reactivity between the surface antigens of the two viruses in immunodiffusion experiments suggested that the major antigenic determinants of the viral surfaces are different; however, results of passive hemagglutination tests indicated that there are common minor determinants. Nucleic acid homology, as measured by liquid hybridization, was found to be 3 to 5% of the viral genomes. The results of this study provide further evidence that woodchuck hepatitis virus is the second member of a new class of viruses represented by human hepatitis B virus. Since virus-infected woodchucks may acquire chronic hepatitis and hepatocellular carcinoma, these antigens and their respective antibodies will be useful markers for following the course of virus infection in investigations of the oncogenic potential of this class of viruses. The nucleocapsid antigen described may be a class-specific antigen of these viruses and, thus, may be useful in discovering new members of the group.


Detection of polymorphism in BALB/c leukemia viruses with mouse antisera. Antisera produced in mice recognize primarily type-specific antigenic determinants on both the major core protein, p30, and the major envelope proteins, gp70 and p15(E), of the endogenous leukemia viruses (MuLV) of BALB/c mice. Three different mouse sera were investigated in detail. (i) Antisera prepared in C57BL/6 mice against the AKR leukemia K36 reacted with the gp70, p15(E), and p30 proteins of MuLV. Certain pools of the C57BL/6 anti-AKR K36 serum contained antibodies which serologically distinguished the p30 proteins of N-ecotropic, B-ecotropic, and xenotropic BALB/c MuLV. (ii) Antisera prepared in BALB/c mice against the BALB/c sarcoma 1315 contained antibodies that reacted with a type-specific antigen of the 1315 MuLV gp70 that is not found on other BALB/c MuLV. (iii) The normal sera of multiparous BALB/c mice contained antibodies that reacted with gp70 and p15(E) proteins of ecotropic MuLV. Sera from some of these mice contained antibodies that serologically distinguished the gp70 of N-ecotropic and B-ecotropic BALB/c viruses. These results emphasize the utility of mouse antisera in the serological typing of MuLV. Furthermore, the antigenic differences observed in the p30 and gp70 proteins should be of particular use in the future analysis of recombinant BALB/c MuLV.


Pheasant virus DNA polymerase is related to avian leukosis virus DNA polymerase at the active site. The DNA polymerase from Amherst pheasant virus (APV), a member of the pheasant virus species of retroviruses, was compared to the DNA polymerases of avian leukosis viruses (ALV) and a reticuloendotheliosis virus (spleen necrosis virus (SNV)). Immunoglobulin inhibition tests and competition immunoassays showed that APV and ALV DNA polymerases are closely related at their active sites. The determinants common to their active sites are not shared by SNV DNA polymerase. Bu using a species-specific radioimmunoassay, it was shown that both APV and SNV DNA polymerases are grossly different from ALV DNA polymerase. The specificity of the relationship of the active sites of APV and ALV DNA polymerases was confirmed by a heterologous radioimmunoassay. Our data indicate that pheasant viruses are evolutionarily linked to ALV.


Isolation of basic protein-acidic lipid complex from myelin. When lyophilized myelin prepared from canine brain white matter was extracted with a solvent mixture of n-propanol-n-hexane (2:3, v/v), it was found that the extract contained less amounts of sulfatide and phosphatidylserine and no protein like so-called proteolipid-protein in comparison with the total chloroform-methanol (2:1, v/v) extract of myelin. Thus, the propanol-hexane myelin residue was re-extracted with chloroform-methanol (2:1, v/v) and the extract was subjected to UV-spectroscopy, TLC and slab SDS-gel electrophoresis, respectively. The UV-absorption pattern of the extract showed a maximum band due to protein at about 280 nm. The lipid portion was found to be predominantly sulfatide (63.6%), phosphatidylserine (26.2%) and other lipids (10.2%), while the major portion of protein was basic protein. The amino acid composition of the extract was also very similar to that of a control porcine basic protein. The major normal fatty acids of sulfatide were C24:0 (34.5%), C24:1 (13.7%), C23:0 (8.3%) and C18:0 (16.0%), while the hydroxy acids were predominantly C24:0 (44.1%), C24:1 (22.2%) and C23:0 (10.4%). Also, the fatty acids of phosphatidylserine were mainly C18:0 and C18:1. The solubilization of the basic protein-acidic lipid complex (about 2.56 by weight in the ratio of protein to lipid) in chloroform-methanol suggested the interaction of the basic protein with acidic lipids which might be related to the molecular organization in myelin. The basic protein-acidic lipid complex was also found to have an encephalitogenic activity which seemed to be a little different from that of basic protein itself.


Changes in cardiac norepinephrine in spontaneously hypertensive and stroke-prone rats. The norepinephrine (NE) concentration of cardiac ventricles was determined by radioenzymatic assay in normotensive Wistar Kyoto rats (WKY), spontaneously hypertensive rats (SHR), and stroke-prone rats (SPR) at 3-6, 14-19, and over 31 weeks of age. There was no difference between strains prior to hypertension, but a progressive decrease in cardiac NE concentration occurred in SHR and particularly in SPR relative to WKY after hypertension was established. This decrease was not due to cardiac hypertrophy. The cardiac neuronal NE storage capacity in rats over 31 weeks of age was analyzed by determining the maximum concentration of NE obtained in a cardiac microsomal fraction, after saturation in vivo with exogenous NE. The results indicated that, after a long period of hypertension, there was a reduction in cardiac NE storage capacity resulting from a loss either of sympathetic nerve endings or of storage vesicles. Moreover, in addition to this reduction in the total size of the cardiac NE store, there was an independent reduction in the degree of filling of this store in both SHR and SPR. This could reflect an increased turnover of cardiac NE in chronically hypertensive SHR and SPR.


Characterization of the coronary vasodilator and hemodynamic actions of monensin, a carboxylic ionophore. The effects of monensin on coronary blood flow (CBF) and other hemodynamic parameters were studied in anesthetized dogs. A dose-response relationship was established, and it was found that the lowest doses of monensin (5-25 micrograms/kg) produced a dose-dependent increase (3-5x) in CBF with a concomitant decrease in total peripheral resistance (TPR). Pretreatment with diphenhydramine, atropine, indomethacin, or propranolol resulted in no reduction in peak increase in CBF or in the duration of response to monensin. However, the response was partially blocked by aminophylline. Large doses (100 and 200 micrograms/kg) produced a dose-dependent increase in cardiac output, aortic pressure, and LV dP/dt max. The duration of these effects was dose-dependent, ranging from 60 to 120 min or longer. Heart rate remained unchanged with all doses of monensin. Pretreatment with propranolol, H87/07 (a cardioselective beta-blocker), and D-600 given alone or in combination significantly reduced, but did not completely abolish, the monensin-induced increase in LV dP/dt max and aortic pressure responses. The increase in CBF in the left anterior descending coronary artery was not significantly affected by these drug pretreatments. Thus, our studies indicate that monensin has two distinct pharmacological effects--in the lower dose range (less than 25 micrograms/kg) it produces a direct relaxation of the blood vessels resulting in an increase in CBF and a decrease in TPR; at high doses (greater than 25 micrograms/kg) it increases myocardial contractility and aortic blood pressure.


Beneficial actions of bevantolol on subendocardial blood flow and contractile function in ischemic myocardium. The effect of a new cardioselective beta adrenergic antagonist, bevantolol (CI-775), on regional myocardial blood flow and contractile function distal to a severe flow-limiting stenosis of the left circumflex coronary artery was studied in open-chest dogs. Bevantolol (1 mg/kg, i.v.) or saline was administered 30 min after production of left circumflex stenosis sufficient to reduce resting coronary blood flow and contractile force approximately 40%. Regional myocardial blood flow and contractile force were measured with radiolabeled microspheres and Brodie-Walton strain gauge arches, respectively. No significant changes were observed in the saline-treated group. Following bevantolol treatment subendocardial blood flow (1.30 +/- 0.29 to 0.93 +/- 0.19 ml/min/g) and contractile force decreased (11.4 +/- 4.4%) significantly (p less than 0.05) in nonischemic myocardium. Subendocardial blood flow (0.59 +/- 0.14 to 0.81 +/- 0.14 ml/min/g) and contractile force increased (29.3 +/- 3.6%) significantly (p less than 0.05) in ischemic myocardium. These results suggest that bevantolol produces a favorable redistribution of flow to ischemic subendocardium. The increase in flow results in an improvement of contractile function in the ischemic region.


Reversal of serotonin vasodilatation in the dog external carotid bed by sympathetic denervation. In view of the conflicting reports of both constrictor and dilator effects of serotonin on the external carotid vascular bed of dogs, the influence of intraarterial infusions of the amine on blood flow through this territory was assessed by electromagnetic flowmeter techniques. In anesthetized intact dogs, serotonin produced dose-related increases in flow. These vasodilator responses were markedly diminished after ipsilateral vagotomy and were followed by delayed vasoconstriction, which then became the predominant response. Pretreatment with atropine did not modify dilator responses, while resection of the ipsilateral stellate ganglion reversed them to pure constriction. In the internal carotid, serotonin elicited constriction, and this effect was unaffected by vagotomy. These results were interpreted in terms of the hypothesis relating serotonin reactivity to vascular tone. The amine would elicit dilatation through an effect on tonically constricted small vessels. Removal of this tone by section of the vagosympathetic trunk or stellectomy would unmask the constrictor effect of serotonin on large vessels. Such reversal of responses would not occur in the internal carotid, a territory normally devoid of important sympathetic tone.


Myocardial procainamide concentration in canine atria and ventricles. The aim of this investigation was to examine the distribution of procainamide in the canine heart. Eight anesthetized open-chest dogs received intravenous infusions of 14C-labeled procainamide at 40 micrograms/kg/min for 4 hr. Just prior to the end of the infusion period, 51Cr-labeled microspheres were injected into the left atrium to measure regional myocardial blood flow. The heart was then excised and dissected into regional myocardial sections from each cardiac chamber. Samples from each section were combusted, and the liberated 14CO2 was trapped and counted to determine regional myocardial procainamide concentration. The remainder of each section was analyzed for 51Cr to determine regional myocardial blood flow. Plasma procainamide concentration (mean +/- SE) at the termination of the infusion was 2.68 +/- 0.14 micrograms/ml. Left ventricular procainamide concentration was 6.09 +/- 0.46 micrograms/g. The right ventricular drug concentration was 94% (NS), the left atrial concentration was 85% (p less than 0.005), and the right atrial concentration was 79% (p less than 0.005) of the left ventricular concentration. Within the myocardium of each cardiac chamber there was no correlation between drug concentration and blood flow. We conclude that these concentration differences are insufficient to explain differences in procainamide efficacy between ventricular and atrial arrhythmias.


Mechanism of antihypertensive activity of orally administered prazosin in spontaneously hypertensive rats. In conscious, spontaneously hypertensive rats (SHR) oral prazosin (0.03-3.0 mg/kg) resulted in dose-related reductions of systolic blood pressure measured with a tail cuff. In SHR whose tail artery blood pressure was continuously monitored the antihypertensive effect of prazosin (1.0 mg/kg, p.o.) was accompanied by a significant increase in heart rate. Several groups of SHR were pithed 2 hr after oral prazosin (1.0 mg/kg) or placebo. In this preparation the mean carotid blood pressure increases following i.v. injections of angiotensin II or 5-hydroxytryptamine and the positive chronotropic responses to i.v. norepinephrine or electrical stimulation of the spinal cord were similar in control and prazosin-pretreated animals. However, the dose-pressor response curves to i.v. norepinephrine or electrical stimulation of the spinal cord from prazosin-pretreated SHR lay to the right of the control curves. In addition, the slopes of the linear portion of these curves were flatter after prazosin and remained so even after i.v. propranolol was given alone or with cocaine. Prazosin-pretreated SHR responded to phenylephrine with a fall followed by a rise in carotid blood pressure. The depressor effect was abolished and the pressor phase enhanced by i.v. propranolol. The pressor responses to i.v. cirazoline or clonidine were significantly inhibited by prazosin. Finally, prazosin failed to significantly modify the negative chronotropic effects of clonidine observed in pithed SHR whose heart rate was raised by continuous electrical stimulation of the thoracic spinal cord. These results indicate that oral prazosin exerts pronounced antihypertensive effects in the SHR. This action apparently results from impairment of the sympathetic nervous system at the level of vascular postsynaptic alpha-adrenoceptors.


Mexiletine in the prophylaxis of ventricular arrhythmias during acute myocardial infarction. In a double-blind study involving 165 patients we examined the role of mexiletine, a new antiarrhythmic drug, for the prophylaxis of ventricular arrhythmias after acute myocardial infarction. Mexiletine or placebo was given orally to patients on arrival in the coronary care unit, and continuous electrocardiographic tape recordings were used to document arrhythmias. Ventricular tachycardia and R-on-T ventricular ectopic beats were significantly reduced in the mexiletine patients, but too few episodes of ventricular fibrillation occurred for statistical comment. When arrhythmias did occur in the mexiletine group, it was usually early in the study, at which time plasma drug levels were low. Adverse effects were uncommon. Patients who were given therapy, but in whom acute myocardial infarction could not be confirmed, suffered no serious consequences of taking mexiletine. The results demonstrate the benefit and limitations of prophylactic oral antiarrhythmic therapy for patients with acute myocardial infarction.


Effects of phenytoin on refractoriness and conduction in the human heart. Using His bundle electrograms and the atrial (A2) and ventricular extrastimulus (V2) techniques, anterograde and retrograde refractory period studies were performed (in 9 and 12 patients, respectively) before and 10 min after intravenous infusion of phenytoin (DPH; mean plasma level, 17.3 micrograms/ml). DPH had no effect on the duration of the QRS complex or the H-V interval of the sinus beats; it had variable but insignificant effects on the sinus rates and the atrial, A-V nodal, and ventricular muscle refractoriness. With the use of the A2 technique, the effective refractory period (ERP) of the His-Purkinje system (HPS) could not be determined in any patient; the relative refractory period (RRP) of the HPS could be determined in 2/9 patients and shortened in both patients after DPH. With the use of the V2 technique, retrograde functional refractory period (FRP) and RRP of the HPS could be determined in all 12 patients and the retrograde ERP of the HPS in 7/12; DPH significantly shortened all these parameters (p less than 0.001, less than 0.001, and less than 0.005, respectively). Functional refractory period of the ventriculo-atrial conduction system (VACS) could be determined in 11/12 patients during control studies (the remaining one patient had complete ventriculo-atrial block). DPH significantly shortened the FRP of the VACS in those (4) patients (Group I) in whom it was determined primarily by the HPS (p less than 0.025), and had variable but insignificant effects on FRP of the VACS in the other seven patients (Group II) in whom it was determined almost exclusively by the A-V node. DPH significantly decreased the retrograde HPS conduction times of the premature impulses (V2H2 intervals) for the same coupling (V1V2) intervals (p less than 0.001). It is concluded that, in the human heart, DPH exerts its most important effects on the HPS where it significantly decreases refractoriness and enhances conduction of the premature impulses. This study also demonstrates that the V2 technique is far superior to the A2 technique for evaluating the effects of drugs on refractoriness and conduction in the HPS.


Methylprednisolone sodium succinate treatment in global ischemia of the cat isolated heart. Effects of methylprednisolone were studied on isolated, blood-perfused cat hearts subjected to 1 hr of normothermic ischemic arrest. Untreated hearts sustained decreases in peak ventricular pressure pulse, dP/dt, and ventricular compliance. Ischemic hearts also became edematous, gained sodium, and lost potassium and creatine kinase enzyme activity. Steroid treatment did not significantly alter any of these ischemia-induced changes. Methylprednisolone treatment did increase resting coronary flow and also increased the hyperemic response after reperfusion. These results, in isolated hearts, provide no evidence that steroid treatment exerts a direct protective effect on the globally ischemic myocardium.


Paradoxical actions of quinidine in hyperpermeable heart cells. The effect of quinidine on the inotropic state was examined using small fragments of hyperpermeable ventricular muscle in vitro. In concentrations of 10(-5) and 10(-4) M, quinidine had no effect on the force of spontaneously contracting fibers. At a concentration of 10(-3) M, contractile force increased by at least twofold. We conclude that the negative inotropic action of quinidine is not by a direct action on intracellular organelles of the heart.