Datasets:

andersonbcdefg

/

pubmed_pairs_part2

like 0

Binding properties of immunoglobulin combining sites specific for terminal or nonterminal antigenic determinants in dextran.

Binding constants of the dextran-reactive BALB/c mouse IgA myeloma proteins W3129 and QUPC 52 have been determined for each member of the isomaltose series of oligosaccharides and for methyl alphaDglucoside. Protein W3129 has maximum complementarity for isomaltopentaose (IM5) deltaf degrees = 7,180 cal/mol) with 55-60% of the total binding energy directed against methylalphaDglucoside. Protein QUPC 52 gives maximum binding with isomaltohexaose (IM6) (deltaF degrees = -5,340 cal/mol) and has about 70% of its total binding energy for isomaltotriose (IM3), but at most only 5% for isomaltose (IM2) or methyl alphaDglucoside. Protein W3129 precipitates with branched dextrans high in alpha (1 yields 6) linkages and reacts with but does not precipitate a synthetic alpha (1 yields 6)-linked linear dextran. Protein QUPC 52 precipitates both branched and linear dextrans. Thus, the immunodominant group for protein W3129 is mimicked by methyl alphaDglucoside and this protein reacts exclusively at the terminal nonreducing ends of alpha (1 yields 6)-linked dextran chains. Protein QUPC 52 has an immunodominant group which is expressed by IM3 but not smaller oligosaccharides and this protein can react at nonterminal locations along alpha (1 yields 6)-linked dextran chains. Precipitation of linear dextran seems to be a valid although not quantitative assay for antidextrans with nonterminal specificity. Quantitative precipitin reactions with branched and linear dextrans suggest that alpha (1 yields 6)-specific human antidextrans are mixtures of molecules having terminal and nonterminal specificities and that the fraction of each type can vary among individuals. Rabbit antisera against IM3 or IM6 coupled to bovine serum albumin also appear to contain antibodies with nonterminal specificity for dextran chains although a large fraction has terminal specificity. Low molecular weight clinical dextran N-150N (congruent to 60,000) reacted more like linear dextran than like its parent native-branched dextran B512. This is thought to result from an abundance of nonterminal determinants in clinical dextran N-150N but a very small number of functional terminal determinants per molecule. An appreciation of terminal and nonterminal specificities and of the different immunodominant structures in isomaltosyl chains has proven to be of a great value in understanding the immunochemical reactions of dextrans. Moreover, certain previous findings with fructosan-reactive mouse myeloma proteins and human antilevans (55, 84) also suggest terminal and nonterminal specificities for levan chains.

The fine structure of hyaline inclusions (pseudopsammoma bodies) in meningiomas.

"Hyaline inclusions" of a meningothelial meningioma were examined under the electron microscope. These eosinophilic, PAS-positive proteinaceous structures, which under the light microscope are seen in both extra- and intracellular location, were found by electron microscopy to consist of granular masses surrounded by cell membranes with microvilli. The "intracellular" bodies were also surrounded by similar microvilli in a space within the cells. Such spaces, variously known as intracellular ductules and "neolumen formation", have been previously described in mammary cancer, bronchiolar carcinoma and pleural mesotheliomas, among others, and have been, in the latter instances, regarded as signs of secretory differentiation. Thus, hyaline inclusions of meningiomas are different from truly intracellular hyaline bodies of neoplastic astrocytes (found by Rubinstein and Herman to be bodies within autophagic vacuoles) and may be regarded as a possible factor in ultrastructural differential diagnosis between meningiomas and gliomas.

Depressed immune response in the magnesium-deficient rat.

The effects of dietary magnesium on growth, food efficiency, organ development, splenic nucleic acids, and serum antibody were studied in two experiments with male Wistar rats. Diets containing 30% protein from casein were fed ad libitum. Rats were immunized intravenously with sheep red blood cells. Blood was obtained 5 and 9 days after immunization. In experiment 1, a group of weanling rats was fed 10 ppm Mg for 8 days, followed by 142 ppm for 37 days. Group 2 (controls) was fed 480 ppm Mg for 45 days. Group 1 weighed less but had larger spleens, kidneys, and testes relative to body size than did group 2. Nucleic acids per gram spleen were similar in both groups as were serum gamma-globulin and its 19S and 7S components. Antibody log titers for group 1 were 45 and 65% of control agglutinin levels and 44 and 80% of control hemolysin values on days 5 and 9, respectively. In experiment 2,200-g rats were fed 10 (group 3) or 480 ppm (group 4) Mg for 38 days. Most effects of the 10 ppm Mg diet were similar to those seen in magnesium deficiency in experiment 1. Antibody titers for group 3 were 30 and 25% of control agglutinin and 43 and 53% of control hemolysin values on days 5 and 9, respectively. Total serum gamma-globulin and its 19S fraction were similar in both groups, while the 7S fraction of group 3 was only 64% of the control value.

Membrane potentials in pinched-off presynaptic nerve ternimals monitored with a fluorescent probe: evidence that synaptosomes have potassium diffusion potentials.

1. Some physiological properties of tissue fractions from rat brain homogenates have been examined. Of the three fractions studied (presynaptic nerve terminals, mitochondria and fragmented membranes), only the nerve terminals (synaptosomes) have the ability to accumulate 42K from physiological salt solutions. 2. The ability to accumulate and retain K is lost if synaptosomes are exposed to very hypotonic solutions. The K uptake and total K content is reduced by ouabain and by inhibitors of glycolysis and oxidative phosphorylation. 3. These results suggest that synaptosomes in physiological saline accumulate K against a concentration gradient, and may have K diffusion potentials across their surface membranes. The voltage-sensitive fluorescent probe, 3,3'-dipentyl 2,2'-oxacarbocyanine (CC5), was used to test this possibility. 4. In the squid axon, the fluorescent emission of CC5 is directly proportional to membrane potential; depolarization causes an increase in fluorescence. 5. The fluorescence of synaptosomes ('synaptosome fluorescence') treated with CC5 is increased when [K]o is increased or [K]o is reduced; replacement of external Na by Li or choline has little effect on the synaptosome fluorescence. In quantitative terms, synaptosome fluorescence is proportional to log ([K]o plus 0-05[Na]o). Rb is about as effective as K in enhancing synaptosome fluorescence; Cs is about 1/4 as effective. The effect of increased [K]o is reversible. 6. The fluorescence data provide corroborative evidence that there is normally a large K gradient ([K]o smaller than [I]i) across the synaptosome surface membrane. The data suggest the [K]i may be in excess of 100 mM. 7. Replacement of Cl- by methylsulphate did not significantly affect the relationship between synaptosome fluorescence and [K]o, nor did removal of external Ca. 8. The fluorescence of CC5-treated mitochondria, membrane fragmnets, or lysed synaptosomes is unaffected by changes in the K concentration of the medium. 9. Veratridine and gramicidin D, both of which enhance Na permeability (PNa) in some intact tissues, increase synaptosome fluorescence when added to the standard medium. The increment is greatly reduced or abolished when external Na is replaced by choline. 10. If synaptosomes are first Na-loaded (by pre-treatment with cyanide + iodoacetate), and then placed in a choline medium, addition of gramicidin D significantly decreases fluorescence. This effect could be explained if, with [Na]o smaller than [Na]i, the increase in PNa causes the synaptosomes to hyperpolarize. 11. The veratridine-induced increase in synaptosome fluorescence was prevented by 3 times 10- minus 7M tetrodotoxin, which also blocks the depolarizing effect of veratridine in intact neurones. 12. The main conclusion is that synaptosomes may retain resting membrane potentials and the ability to increase Na permeability.

Structural proteins of mammalian RNA tumor viruses: relatedness of the interspecies antigenic determinants of the major internal protein.

The relatedness of antigenic determinants of purified major core proteins of the murine, feline, RD 114/baboon, and woolly monkey/gibbon ape groups of RNA tumor viruses was examined by competition radioimmunoassay. In assay systems of a homologous antigen and antiserum, high affinity competition for binding to all of the antibodies was observed only with the homologous unlabeled protein; the core proteins of other groups of viruses showed only low affinity binding of a small fraction of antibodies, presumably those reactive with the interspecies determinants, at concentrations of competing protein 10- to 100-fold greater than that of the labeled antigen. The cross-reactive (interspecies) antigens of every two viruses were selectively examined by precipitating the purified 125-I-labeled protein with antiserum against each of the other proteins. The extent to which these shared determinants were common to the other viruses was then tested by the effectiveness of the proteins of each virus to compete for antibody binding. Several classes of interspecies determinants were distinguished: those common to two of the groups of viruses, others to three, and some to all four. Moreover, an even greater variety of interspecies determinants was indicated by differences in the affinity of the individual proteins for antibody binding, supporting the hypothesis that there are at least several, if not many, different interspecies determinants with a broad spectrum of antigenic cross-reactivity. These studies suggest that the murine and feline viruses are closely related as they contain cross-reactive antigenic determinants not shared with the other viruses, that the feline virus is more closely related to the woolly monkey virus than to RD 114, and that the RD 114 and woolly monkey viruses retain interspecies determinants shared relatively equally with each of the other viruses.

Interspecies antigenic determinants of the reverse transcriptases and p30 proteins of mammalian type C viruses.

The major internal structural proteins (p30) of type C viruses isolated from several mammalian species were studied by radioimmunoprecipitation and competitive radioimmunoassays. Three antigenically distinguishable sets of interspecies determinants could be demonstrated by both methods. One set of determinants shared by viruses of rodent origin (mouse and rat) can be detected readily in feline leukemia viruses but not in other type C viral groups. The p30 proteins of murine viruses also contain a second discrete set of antigenic determinants related to those in infectious primate viruses and endogenous porcine viruses, but not detected in the feline leukemia virus group. The p30 proteins of endogenous viruses of baboons and domestic cats share yet a third set of cross-reactive determinants not detected in type C viruses isolated from other species of animals. Enzyme inhibition studies performed with antisera raised toward the reverse transcriptases of these same groups of type C viruses showed the same patterns of immunological cross-reactions as observed with p30 proteins. The antigenic cross-reactions between the homologous proteins of type C virus isolated from genetically distant animals may reflect transmission of type C viruses across species barriers.

Gut bacteria and their metabolic activities in familiar polyposis.

Earlier work had suggested that patients with large-bowel cancer can be characterized by carriage of clostridia capable of dehydrogenating the nucleus of steroids and by high faecal bile-acid concentrations. Familial polyposis is an inherited disease which untreated, will progress to cancer of the large bowel, and those affected might be expected to have these metabolic characteristics. However, enviromental factors seem to play no part in polyposis. Investigation of as yet unaffected children of known polyposis patients revealed that the gut flora of half of them did not degrade cholesterol in vivo andnor did flora from patients in whom polyposis had already been diagnosed. The reason for this is unknown, but if the same patients develop polyposis (i.e., they carry the abnormal gene) we would have a simple diagnostic test which could replace the continuing follow-up now required for the siblings and children of patients with polyposis.

Role of prostaglandins in the pathogenesis of acute renal failure.

It is suggested that impaired glomerular infiltration in acute renal failure may be due, if only in part, to the derangement of a normal local feedback mechanism involving prostaglandins. According to this hypothesis, prostaglandins made in the medulla normally enter the loop of Henle and thus are present in distal tubule fluid reaching the macula densa. In the absence of direct vascular channels connecting the renal medulla with the outer cortex, this may be the only route by which medullary prostaglandins can reach the cortex without being destroyed by dehydrogenases. Under most circumstances in which renal perfusion and glomerular filtration fall, flow through Henle's loop continues, albeit at a reduced rate. This slowed flow through the loop and augmented prostaglandin release combine to produce a high concentration of prostaglandin in fluid leaving the medulla which, acting on the macula densa, counteracts the vasoconstrictor stimulus. If, however, filtration stops completely because of overwhelming vasoconstriction, flow along the loop also stops ans prostaglandins no longer reach the macula densa. Thus, the prostaglandin feedback loop would be opened, constrictor mechanism remain unchecked, and filtration failure perpetuated. It is proposed that such a self-perpetuating mechanism might operate in acute renal failure.

Prevention of fatal postoperative pulmonary embolism by low doses of heparin. An international multicentre trial.

The efficacy of low-dose heparin in preventing fatal postoperative pulmonary embolism has been investigated in a multicentre prospective randomised trial. 4121 patients over the age of forty years undergoing a variety of elective major surgical procedures were included in the trial; 2076 of these were in the control group and 2045 patients received heparin. The two groups were well matched for age, sex, weight, blood-group, and other factors which could predispose to the development of venous thromboembolism. 180 (4-4 %) patients died during the postoperative period, 100 in the control and 80 in the heparin group: 72% of deaths in the control and 66% in the heparin group had necropsy examination. 16 patients in the control group and 2 in the heparin group were found at necropsy to have died due to acute massive pulmonary embolism (P smaller than 0-005). In addition, emboli found at necropsy in 6 patients in the control group and 3 in the heparin group were considered either contributory to death or an incidental finding since death in these patients was attributed to other causes. Taking all pulmonary emboli together, the findings were again significant (P smaller than 0-005). Of 1292 patients in whom the 125-I-fibrinogen test was performed to detect deep-vein thrombosis (D.V.T.) 667 were in the control group and 625 in the heparin group. The frequency of isotopic D.V.T. was reduced from 24-6% in the control group 7-7% in the heparin group (P smaller 0-005). In 30 patients D.V.T. was detected at necropsy; 24 in the control and 6 in the heparin group (P smaller 0-005). 32 patients in the control group and 11 in the heparin group developed clinically diagnosed D.V.T. which was confirmed by venography (P smaller than 0-005). In addition, 24 patients in the control and 8 in the heparin group were treated for clinically suspected pulmonary emoblism. The difference in the number of patients requiring treatment for D.V.T. and/or pulmonary embolism in the two groups was again significant (P smaller than 0-005). 9 patients were found at necropsy to have died from haemorrhage; 5 were in the control and 4 in the heparin group. A careful objective analysis of operative and postoperative bleeding in 1475 patients showed no statistically significant difference in the blood-transfusion requirements or in the fall in the postoperative haemoglobin level either in the individual operative groups or in the group as a whole. However, the difference in the number of patients who developed wound haematoma in the heparin and control groups was significant (P smaller 0-01). The results of the trial indicate that this form of prophylaxis can now be recommended for use on a large scale in "high-risk" patients undergoing major surgery.

Cardiac and pulmonary effects of acebutolol.

In a double-blind randomised study, single intravenous doses of propranolol (0-1 mg. per kg.), practolol (1 mg. per kg.), acebutolol (1 mg. per kg.), or placebo were each administered at weekly intervals to six healthy volunteers. Forced expiratory volume in 1 second (F.E.V.1), resting and exercise heart-rate, and resting and exercise peak flow-rate (P.F.R.) were determined before and at 2, 3, 4, and 6 hours after each treatment. Venous blood-samples were also obtained at these times. Compared with placebo, resting heart-rate was reduced after all three drugs, but the corresponding differences in exercise heart-rate were much greater, more consistent, and of greater statistical significance. At 2, 3, and 4 hours when acebutolol and propranolol produced equivalent cardiac beta-blocking activity (judged by reductions in exercise heart-rate), their mean plasma concentratios were in the ratio of about 8/1; and at 2 hours when practolol and acebutolol gave rise to almost equivalent cardiac beta blockade, their mean plasma concentratio ration was 3/1. At times, reductions in F.E.V.1 and resting P.F.R. after propranolol (but not after practolol or acebutolol) were significantly greater than the corresponding changes after placebo. The reductions in exercies P.F.R. after propranolol (6 hours) and acebutolol (4 hours) (but not after practolol) were significantly greater than the changes after placebo. Changes in F.E.V.1, resting and exercise P.F.R. after propranolol, and the corresponding changes after practolol, were significantly different, all of which confirmed that practolol was more cardioselective than propranolo. In general, the reductions in F.E.V.1 and resting P.F.R. after acebutolol were slightly smaller than after propranolol, excepting at 6 hours when the difference between them was significant. The reductions in exercise P;F.R. after acebutolol and propranolol were of the same order, there being no significant differences between the two, whereas the reductions after acebutolol were clearly greater than the corresponding changes after practolol, the differences being significant at 2, 3, and 4 hours.

Diarrhoea associated with heat-stable enterotoxin-producing strains of Escherichia coli.

Five patients who developed acute watery diarrhoea while travelling in Mexico in October, 1974, were found to have enterotoxigenic Escherichia coli in their stool which produced heat-stable enterotoxin (S.T.) without producing heat-labile enterotoxin (L.T.). These S.T.-only E. coli, which have previously been described as causing diseases in animals, must now be regarded as pathogenic for humans as well.

Effect of phenethyl alcohol and other organic substances on cellulas production.

Cellulase can be produced from growth in noncellulosic substrate if the growth rate of the producing organism is restricted. Phenethyl alcohol (PEA) is a growth inhibitor and was used to control the growth of M. verrucaria in attempts to obtain increased cellulase production. Cellulase yield was found to be increased without a restriction in growth rate when PEA was present in low concentrations (0.03% v/v). The effect was observed for other organisms but notably L. trabea, which produced considerable enzyme from a small quantity of mycelium. Here increased cellulase synthesis was concomitant with restricted growth. Other chemicals with PEA-like structure (e.g. benzyl alcohol) resulted in similar or more extensive cellulase synthesis. Of the substances tried, propyl alcohol was most effective, followed by acetone. PEA causes a swelling of cell walls and inhibits spore formation. This and other data given suggest that PEA affects the cytoplasmic membrane or the cell wall or both. Cellulase synthesis is considered to take place in the membrane and wall region of the cell.

Anomalous results of studies on drug interaction in man. I. Nortriptyline and antipyrine.

Conflicting results were obtained in three drug interaction experiments designed to determine in man effects of chronic nortriptyline administration on plasma antipyrine half-lives. In an apparently similar design, each experiment utilized normal volunteers whose plasma antipyrine half-lives were measured before and after chronic nortriptyline administration. The same dose of liquid nortriptyline (0.2 mg/kg p.o. t.i.d. for 8 days) was used in each experiment. All subjects in the first study prolonged their plasma antipyrine half-lives, whereas seven of nine subjects in the second study significantly shortened their plasma antipyrine half-lives. In the third experiment seven of nine subjects showed no significant change in plasma antipyrine half-life. An explanation for these conflicting results obtained in the same laboratory under apparently similar experimental conditions was sought in the nature of the volunteers and in subtle changes in such variables as differences in the batches and length of storage of liquid nortriptyline.

B-cell mitogenic effects on human lymphocytes of rabbit anti-human beta 2-microglobulin.

Rabbit Ig against human beta2-microglobulin was found to be mitogenic for human peripheral lymphocytes, tonsil lymphocytes, and appendix and spleen cells. Anti-beta2-m gave increased DNA synthesis, with peak responses on day 3 or 4 and showed a dose-response effect when diluted. The effect was seen in both serum-free and serum-containing culture medium. Anti-beta2-m, as well as lipopolysaccharide, induced polyclonal antibody production and intracellular immunoglobulin synthesis in blast cells, which is taken as evidence that these substances are human B-cell mitogens. Anti-beta2-m, but not lipopolysaccharide, could, in these experiments, activate peripheral blood lymphocytes, in addition to lymphoid cells from other sources. Thus, anti-beta2-m can serve as a functional marker for peripheral human B lymphocytes.

Stains for A, B, and D cells in fetal rat islets.

Ten techniques often used for identification of A, B, and D cells in adult islets of Langerhans were applied to fetal rat pancreas. Modifications were tried with many of these techniques. Two indole methods (xanthydrol and postocoupled benxylidene reactions) and a cryostat technique using o-phthaladehyde failed to stain fetal islets. Phosphotungstic acid hematoxylin and lead hematoxylin lightly stained fetal A cell granules in Helly's fixed tissue. The Grimelius silver nitrate technique stains adult rat A cells but failed to stain fetal cells. A modification of this technique stained fetal A cells and a possible 4th cell type. The specificity of this method was confirmed by restaining stained cells with a fluorescent antibody technique and with pseudoisocyanin. B cells, as previously reported, were readily stained by the aldehyde fuchsin technique. Fetal D cells were not stained by the Hellerstrom-Hellman alcoholic silver nitrate method, nor did they display pseudoisocyanin metachromasia after acid hydrolysis; they did fluoresce brightly with this technique when viewed with UV light. It was thus possible to distinguish the three usual cell types, plus a possible fourth type, in the fetal rat pancreas.

[Inhibition of viral reverse transcriptase and leukemogenesis by modified nucleic acids (author's transl)].

Inhibition of DNA polymerase from oncorna viruses by a new class of macromolecular inhibitors is reported. The macromolecule, designated as mercaptopolycytidylic acid (MPC), is a chemically modified polycytidylic acid containing 5-SH cytidylic bases in the polymerase. Partially thiolated polycytidylic acids (MPC I-III, containing 1.7%, 3.5%, and 8.6% 5-mercaptocytidylate units, respectively) inhibited the DNA-polymerase of Friend leukemia virus (FVL) in the endogenic reaction as well as in the presence of poly rA-(dT)14 or poly (dA-dT) templates; the inhibitory activities were directly related to the percent of tholation. In a bacterial DNA polymerase (E coli-K12 with denatured calf thymus DNA as template) MPCI-III showed no activity. Biological experiments showed that MPC III inhibits the leukemogenic potential of cell-free spleen extracts from FVL-infected mice to about 60%, measured on the basis of spleen weight. The enzymatic and animal experiments have led us to carry out preliminary clinical trials in some cases of Children leukemia. These cases, resistent to the known therapeutic regimes (combination chemotherapy), responded well when treated with MPC along, or in combination with poly I. The experiments indicate that the development of modified polynucleotids with structural similarities to functional templates may be of potential use in the future chemotherapy of leukemia.

Heterologous protections in experimental salmonellosis.

Immunizations of mice with proteins from S. paratyphi B protected the animals against infection with a concentration of S. paratyphi C which killed the controls and against an infection with 50 LD100 of the homologous S. paratyphi B. The sera of the infected mice showed common precipitation lines of proteins from the two species belonging to different serogroups. Consecutive inoculations with S. typhimurium of both groups of vaccinated mice protected the animals against the infection with their natural pathogen. Immunizations with proteins from S. cholerae-suis protected about 70% of the mice infected with S. paratyphi B and with S. paratyphi C; a higher protection was not, however, induced against infection with the homologous strain. Consecutive infections with S. typhi-murium of the mice resulted in total protection of the animals previously inoculated with S. paratyphi B and S. paratyphi C; the group of mice infected with S. cholerae-suis was less protected against the subsequent inoculation with S. typhi-murium (about 50%). In the infections of mice, S. paratyphi B and S. paratyphi C, seem related to S. typhi-murium by common proteins; the proportion of common protective determinants is not yet known.

Thyroid disease in pregnancy.

It is frequently difficult to establish a diagnosis of hyperthyroidism in association with pregnancy. Signs and symptoms suggestive of hyperthyroidism may occur in normal pregnancy. Moreover, tests to rule out hyperthyroidism under these conditions are not readily available. Mild hyperthyroidism is not a hazard to an otherwise normal pregnancy and does not require therapy on the basis of this presumption. Hyperthyroidism of a clinically significant degree is safely treatable by medical means without hazard to the fetus. Hyperthyroidism is an uncommon cause for the failure of pregnancy to proceed to term. Treatment with thyroid based on a presumption of this diagnosis is justified, but such patients should be studied carefully after delivery to establish the true state of thyroid function for the future. Other conditions of thyroid dysfunction, including thyroiditis, thyroid carcinoma, nontoxic goiter, and ovarian struma rarely interfere with an otherwise normal pregnancy.

The total correction of congenital heart disease in infants.

If radical correction of congenital heart disease can be performed with a low risk in the first year of life the advantages are obvious. Total correction avoids the latent risks of the underlying lesion and the risks of palliative surgery, and relieves the parents and family of the psychological pressure of a major illness. Palliative surgery, although offering survival, may include the long term problems of the palliative operation itself, and the possibility that cases may be lost to further treatment or follow-up examination after a successful palliative operation. For these reasons, to be desirable, a palliative operation should offer a survival chance that is at least 10% better than the corrective procedure in that patient at that time. That is, any corrective procedure with a mortality rate less than 10% is to be perferred at any age to a palliative operation. An accurate assessment of risk demands a complete diagnostic study, and a knowledge of the natural history of that form of congenital heart disease. The problems of infants with congenital heart disease are not primarily caused by age of size. The main problem is that of natural selection.

Evaluation of arrhythmias in the late hospital phase of acute myocardial infarction compared to coronary care unit ectopy.

To evaluate the prevalence and nature of arrhythmias during the entire three-week period in hospital after myocardial infarction, the results of coronary care unit monitoring (initial 3 to 5 days) were compared with continuous 8-hour portable monitoring during the ambulatory phase (second and third weeks) in 83 consecutive survivors. Arrhythmias were detected in 84.3 per cent (70/83) of patients while in the coronary care unit and in 85.5 per cent (71/83) during hospital stay after the coronary care unit. Ventricular ectopic depolarizations were classified as complicated (multifocal, paired, R on T, or five or more a minute) or uncomplicated. Importantly, the high frequency of complicated ventricular extrasystoles and tachycardia persisted during the entire period in hospital (early 34.9% and late 42.5% of all patients). However, only 16.9 per cent (14/83)had these ventricular arrhythmias during both coronary care unit and ward monitoring. Thus, the absence of complicated ventricular ectopic depolarization and ventricular tachycardia in the coronary care unit did not exclude their subsequent occurrence in the majority of the large number of patients with late hospital complicated ventricular ectopy.

Conformational and spectral analysis of the polypeptide antibiotic N-methylleucine gramicidin S dihydrochloride by nuclear magnetic resonance.

The 220-MHz proton magnetic resonance spectrum of the cyclic decapeptide antibiotic, mono-N-methylleucine gramicidin S, is reported and all the resonances have been assigned to specific protons of the constituent amino acids. Three methods--temperature dependence and solvent mixture (methanol-trifluoroethanol and dimethyl sulfoxide-trifluoroethanol) dependence of peptide NH proton chemical shifts and proton deuteron exchange--habe been utilized to delineate peptide NH protons. The results of the above methods, coupled with the observed vicinal alpha-CH-NH coupling constants and chemical shifts, indicate that in trifluoroethanol the peptide NH PROTONS OF D-Phe4, D-Phe9, L-Orn2, and L-Val6 are exposed to the sovent, and those of L-Val1, L-Orn7, and L-Leu8 are solvent shielded and intramolecularly hydrogen bonded. In trifluoroethanol, dimethyl sulfoxide, and methanol, the decapeptide has no C2 symmetry, and there are only minor conformational differences in the different solvents. In the proposed conformation in trifluoroethanol, one-half of the decapeptide retained the hydrogen bonding pattern of gramicidin S, i.e. cyclo-(L-Val1 NH--O-C L-Leu8) (a beta turn) and cyclo-(L-Leu8 NH--O-C L-Val1). The second half of the molecule exhibits a different type of stable beta turn involving the ten-atom hydrogen-bonded ring, cyclo-(L-Orn7-NH--O-C D-PHE4).

The monoclonal nature of lymphatic leukemia.

Monoclonal membrane-bound Ig were found by immunofluorescence on the lymphocytes in the vast majority of cases of chronic lymphocytic leukemia. The distribution of H and L chains among these patients reflected the distribution of surface Ig on normal lymphocytes and IgM was the predominant class. The importance of the study of surface Ig synthesized in vitro is outlined. The simple staining of freshly drawn cells may lead to erroneous conclusions, since an apparently polyclonal staining can result from the anti-IgG antibody activity of a monoclonal surface IgM, from the attachment of immune complexes at the lymphocyte surface or from the binding of serum antibodies to cell membrane determinants. A biclonal proliferation, characterized by distinct surface-Ig markers, was demonstrated in several cases of chronic lymphatic leukemia. Monoclonal surface Ig were also detected on the lymphoblasts in 2 cases of acute transformation of chronic lymphocytic leukemia, in several patients with acute lymphosarcoma cell leukemia and in 1 case of acute lymphatic leukemia. In most cases of acute lymphatic leukemia, the leukemic cells were devoid of detectable B- or T-cell membrane markers. In 2 cases of acute lymphatic leukemia, 1 case of chronic lymphatic leukemia and in all patients with the Sezary syndrome, the leukemic cells appeared to be thymus-derived.

Distribution along the axon and into various subcellular fractions of molecules labeled with (3H)leucine and rapidly transported in the garfish olfactory nerve.

The distribution of molecules labeled with [3H]leucine by fast axoplasmic transport in vivo has been studied in the garfish olfactory nerve after incorporation of the amino acid by the olfactory mucosa. Owing to the size of the nerve, it has been possible to follow the fate of the labeled molecules in 10 different subcellular fractions of 6 consecutive nerve segments. Each segment represents a different part of the profile developed by the transported radioactive molecules. In order to determine the influence of the perikaryon (rate of protein synthesis and rate of protein release into the axon) transport was studied under 3 different conditions: (1) intact nerves (simply labeled with [3H]leucine); (2) nerves cut from the cell bodies 6 h after application of [3H]leucine; and (3) nerves pulse-chase labeled for 1 h. Several conclusions can be drawn. (1) The bulk of the rapidly transported molecules are membranous axonal proteins, as determined by enzyme markers. Most are found in subcellular fractions representing 17% of the total axonal protein. They are synthesized very rapidly in the cell bodies (less than 1 h after isotope deposition) and exhibit the highest specific activities measured. These high specific activities were found in the same axonal membrane fractions in both plateau and crest, suggesting that the membrane precursors are transported as particles rather than as subunits. (2) The majority of these proteins are released into the axon immediately after synthesis; however, at least 30% of the labeled axonal membranous proteins are not released with the fast wave itself but progressively over a long period of time. (3) The majority of the moving material, particularly in membranous fractions, is left behind the fast wave and is deposited in the axon. When the front base of the fast wve has covered 70% of the total nerve length, only 19% of the labeled material of the main axonal membranous fraction appears still to be moving. (4) Proteins with high specific activities are found near the cell bodies and may be the result of early axonal transport of amino acids, diffusing later into the surrounding cells and being incorporated into proteins. Some free amino acids are also transported along the axon.

Binding of bleomycin to DNA in bleomycin-sensitive and -resistant rat ascites hepatoma cells.

The 14C activity of [14C]bleomycin bound to DNA in bleomycin-sensitive rat ascites hepatoma cells (AH-66) was 8.7 times higher than in resistant cells (AH-66F) when the cells were incubated with [14C]bleomycin. The difference in permeability to bleomycin was not significant; uptake of [14C]bleomycin by the sensitive cells was only 1.2 times larger than that by the resistant cells, and the radioactivity incorporated into the nuclei of sensitive cells was only 1.3-fold greater. The bleomycin-inactivating enzyme level in the resistant cells was 3.5 times higher than in the sensitive cells, indicating that the antibiotic incorporated into the resistent cells was reduced in DNA-binding activity to a large extent. The level of protein-free thiol compound in the sensitive cells was 1.8-fold higher than in the resistant cells, suggesting a possible enhancement of bleomycin action by intracellular thiol compound as is found in vitro. These factors probably affect the DNA strand scission and the sensitivity of cells to this antibiotic. Binding of [14C]bleomycin to DNA in vitro was studied in the presence and the absence of dithiothreitol. A large portion of the radioactivity bound in the presence of dithiothreitol was unstable to acid, but the acid-resistant binding was also enhanced by this thiol compound.

Inhibitory and anti-inhibitory factors of rat serum active on the G1-S transition of hepatocyte cell cycle.

Rat hepatocytes are responsive to a serum factor inhibiting their progression through the cell cycle from the late G1 phase to the S phase. After fractionation of normal adult rat serum by two chromatographic steps on DEAE cellulose and sephadex gel filtration, the inhibitory activity was linked to proteins having a high electronegative charge and of apparent high molecular weight. Polyacrylamide gel electrophoretic analysis of active fraction showed that the alpha1 macroglobulin was its main component. Male and female baby rats were sensitive to the inhibitory factor from normal rats. Contrary to the normal adult rat serum the whole hepatectomized adult rat serum did not exhibit any ingibitory activity on the G1-S transition. However, two components having antagonist activities: an alpha1 globulin and a gamma globulin, were separated by chromatographic procedures from hepatectomized rat serum. (a) The alpha1 globulin showed an inhibitory activity. It had an apparent molecular weight lower than that found in normal rats. Its activity was sex related: only male baby rats were responsive. (b) The factor present in the gamma globulin fraction was found to be antagonistic to the alpha1 globulin factor. Its occurrence after hepatectomy explains the absence of inhibitory activity in the serum of hepatectomized rats.

An ultrastructural study of the microfilaments in rat brain by means of E-PTA staining and heavy meromyosin labeling. II. The synapses.

To identify structures involved in the translocation of the synaptic vesicles towards the presynaptic membrane, an ultrastructural study has been undertaken by means of (1) the E-PTA stain and (2) the HMM-labeling procedure. Using serial sections of E-PTA stained nervous tissue, especially those made in transversal and tangential planes, the geometric order of the presynaptic grid and of its constituents has been described in detail. It consisted of dense projections having the shape of small truncated pyramids cut parallel to their hexagonal bases which rested on the electron-lucent presynaptic membrane. The dense projections were arranged at the points of equilateral triangles. Around each dense projection, six asymmetric hexagonal holes were seen to be arrayed in an hexagonal pattern, forming thus the presynaptic sieve. From the spiny tops of the dense projections, which appeared as specialized structures of the dense material coating the inner surface of the plasma membrane at the level of the synaptic cleft, fine filaments, 40--60 A in diameter, radiated and formed a three-dimensional meshwork pervading the presynaptic bag. The dense cytoplasmic coating delineating the plasma membrane served as anchor points for these microfilaments. Upon incubation with rabbit skeletal muscle HMM the microfilaments underwent specific structural changes, consisting of: (1) a striking increase in diameter; (2) the association of periodic and polarized substructures with their surfaces. The synaptic vesicles and mitochondria were seen to be attached to the numerous HMM-decorated filaments or enmeshed in the network formed by these filaments. The actin-like filaments were anchored to the plasma membrane at many points and to the presynaptic dense projections. Following incubation in the buffer alone or in buffer HMM solutions containing Na+ pyrophosphate or ATP, no arrowheaded structures were observed. Thus, a network consisting of actin-like filaments was demonstrated in the presynaptic bag. Of particular interest was the structural relation of the actin-like filaments with the occasional, tapered myosin-like filaments. The role of the presynaptic actin-like network in the transport of synaptic vesicles towards the presynaptic membrane by a mechanism of chemomechanical transduction is discussed. In the postsynaptic dendrite or dendritic spine, a filamentous network was observed to be attached to the subsynaptic web by means of the E-PTA stain and of the HMM-labeling procedure. The occurrence of an actin-like meshwork in the postsynaptic region is suggested to produce changes in the macromolecular configuration of the postsynaptic membrane by a "mechanoenzyme" system similar to that described in the mitochondrial membrane.

The analgesic properties of delta-9-tetrahydrocannabinol and codeine.

The administration of single oral doses of delta-9-tetrahydrocannabinol (THC) to patients with cancer pain demonstrated a mild analgesic effect. At a dose of 20 mg, however, THC induced side effects that would prohibit its therapeutic use including somnolence, dizziness, ataxia, and blurred vision. Alarming adverse reactions were also observed at this dose. THC, 10 mg, was well tolerated and, despite its sedative effect, may analgesic potential.

C banding in polytene chromosomes of Simulium ornatipes and S. melatum (Diptera: Simuliidae).

Polytene and mitotic chromosomes of Simulium ornatipes and S. melatum were subjected to C banding procedures. In both species polytene chromosomes consistently show C banding of centromere regions, telomeres, nucleolar organiser and, unexpectedly, numerous interstitial sites. The interstitial C banding sites correspond to morphologically single polytene bands. Their response is graded and independent of band size. Interstitial C bands in S. ornatipes are scattered throughout the complement, whereas in S. melatum they are clustered. Supernumerary heterochromatic segments in S. ornatipes also exhibit strong C banding and inverted segments can differ from standard in C banding pattern. -- Mitotic chromosomes of both species show a single centric C band with indications of two weak interstitial bands in S. ornatipes, suggesting that many C band regions, detectable in polytene chromosomes, are not resolved by present techniques in mitotic chromosomes. -- Contrary to current opinion that C banding is diagnostic for constitutive heterochromatin, the interstitial C band sites of polytene chromosomes are regarded as euchromatic. Conversely, the heterochromatic pericentric regions of S. ornatipes are not C banded. -- It appears that polytene chromosomes offer a promising system for the elucidation of C banding mechanisms.

"Alpha-pattern coma" in high voltage electrical injury.

Two cases are reported in which reversible deep coma subsequent to high voltage electrical injury occurred in association with alpha frequencies in the EEG. The EEG pattern differs from the alpha rhythm of the normal awake patient by its diffuse distribution and unresponsiveness to a variety of stimulation. The term "alpha-pattern coma" is introduced to designate the occurrence of this pattern in comatose patients. The EEG in the cases described initially demonstrated activity in the alpha frequency which occurred in a generalized distribution and was unresponsive to stimulation. During the early stages of recovery an increased incidence of theta and delta activity was observed. The recovery records contained a low voltage alpha rhythm and responded to photic stimulation. The literature on this subject is reviewed and the possible modes of pathogenesis are discussed. The authors conclude that such examples of alpha-pattern coma are the result of diffuse cerebral damage and might be detected more frequently in comatose patients who survive if these patients are studied earlier in their clinical course with EEGs.

A quantitative comparison of traditional reading of the EEG and interpretation of computer-extracted features in patients with supratentorial brain lesions.

The EEGs of adult patients with suspected supratentorial brain lesions were recorded on paper and on magnetic tape, using a small computer. The spectra of 16 channels were computed on a 40 sec sample. For each channel, a ratio of the type (delta + theta)/(alpha + beta) was computed and displayed on the computer terminal, a measure of the asymmetry in slow wave activity between homologous areas of the head was also displayed. This display is called a canonogram. It is believed to be a meaningful representation of the important characteristics of the EEG in the presence of supratentorial lesions. In order to assess the clinical value of the canonogram, the presumed localization of the lesion obtained from the interpretation of the traditional EEG and from that of the canonogram were compared to the known location of the lesion is a group of 87 subjects. The comparison was made quantitative by the use of a structured report encoding the traditional interpretation and that of the canonogram as well as the reference data (unequivocal surgical, radiological and clinical localizing evidence). The results varied among the anatomical regions: whereas in the frontal and occipital regions the EEG was slightly more accurate than the canonogram, both methods were similar in the temporal areas and the canonogram seemed more accurate in the centro-parietal regions. An attempt was made to interpret these differences. Furthermore, three readers read the cononograms and gave very consistent interpretations. These results show the reliability and value of this simple computer display for the particular type of EEG studied.

The frontal CNV: its dissimilarity to CNVs recorded from other sites.

Two new techniques were used to study the distribution of the CNV recorded from the scalp of humans. One technique was for the suppression of eye movements and the elimination of them fron the average data. The other technique was for the recognition of differences in the form of CNVs recorded from different sites. The latter technique utilized an average vertex CNV as a template pattern against which CNVs from the vertex, left and right frontal, central and parietal sites were matched. The results showed that the frontal CNV was different in form from the vertex, central and parietal Cnvs and that the vertex, central and parietal CnvS were similar in form. An analysis of the amplitudes of CNVs showed that the frontal CNVs were significantly smaller than the CNVs from all other sites, that the vertex CNV was significantly larger than the parietal or central CNVs and that the central CNV was significantly larger than the parietal CNV. There were no differences in amplitude between hemispheres within Condition A responding with the right hand) or Condition B (responding with the left hand) or between Conditions A and B. The results are discussed with respect to their possible theoretical implications.

4-channel 24 hour cassette recorder for long-term EEG monitoring of ambulatory patients.

A 4-channel cassette recorder capable of continuously recording the EEG for 24 h on a C-120 cassette is currently in use at the Montreal Neurological Institute. We have done a limited study on 20 patients with over 2000 h of recording to evaluate the recording system, its limitations and its capabilities, and to develop methodology for its operations. The use of preamplifiers enables one to record the background EEG with a noise level of 3-5 muVpp on 4 channels continuously for 1 day. The electrodes can be hidden in the hair and the preamplifier under the collar, to allow the patient to carry on his normal activities at home and at work. Since the cassette and the batteries can be easily changed, the patient himself can prolong the recording for several days. The cassette can be played back as fast as 60 times on a Minograf EEG machine for compressed analysis or 20 times for more detailed write-out, to obtain a record equivalent to standard EEG recordings. The system was not designed to replace or compete with standard EEG recordings but when used on well defined clinical problems, or in specific research projects, it can enhance the diagnostic or research value of the EEG.

Sleep studies on a 90-minute day.

After 2 adaptation and 2 baseline all-night sleep recordings, 5 normal young adult subjects (3 males) were placed on a schedule alternating 60 min of wakefulness and 30 min of sleep for 5 1/3 24-h periods. A 2-day recovery period followed. One male subject (MA15) was later placed on the identical protocol with the exception that he was allotted periods of 75 min of wakefulness and 15 min of sleep during the experimental period. One male narcolepsy-cataplexy patient was placed on the 60-30 schedule for 48 h. All subjects showed REM sleep during the schedule manipulation. REMM sleep occurred within 10 min of sleep onset (SOREMP) on 79 of 110 REM sleep occasions in the normals, on all 29 REM episodes in MA15, and on 16 of 17 REM periods in the narcoleptic. In the normals, REM sleep showed a tendency to recur on alternate 90-min cycles, while in the narcoleptic REM recurred on consecutive periods. Compared to baseline, REM sleep 24 h was decreased in the normals and increased in the narcoleptic. Time spent in slow wave sleep and stage 2 was also reduced in the normal subjects on the 90-min schedule, and stage 1 sleep time was increased. Peak sleep times for the 5 normals occurred between 09.00 and 12.30 and lowest sleep times from 21.00 to 02.00. During the first recovery night, sleep times ranged from 11.5 to 18.5 h, including significant increases of slow wave sleep and REM sleep. Except for SOREMPs, no signs of the narcolepsy-cataplexy syndrome were seen in any of the normal subjects.

Effects of time and tasks upon auditory and somatosensory evoked potentials in man.

Auditory evoked potentials (AEPs) recorded from the vertex and somatosensory evoked potentials (SEPs) recorded from the somatosensory and motor areas and vertex were examined during visual and auditory discrimination tasks, with and without motor responses, and during motor tasks alone in normal human subjects. These procedures allowed the separation of sensory discrimination from motor activity effects on the evoked potentials. It was observed that EPs were modified systematically by task and by temporal variables, even though vigilance, as evaluated through EEG recording and performance level, was stable throughout the experimental recording session. 1. AEPs were minimally influenced by time, but very sensitive to task. Inversely, the SEP amplitude decreased considerably with time and less with task. There was evidence of time/task interaction. 2. The magnitude of SEP attenuation in time was reduced by intervening rest periods. 3. The sensory modality in which the discrimination task was performed did not influence the effect on EPs. A discrimination task involving a motor response reduced EPs more than a pure discrimination or a pure motor task. The task effect seems to involve a general, mechanism (load imposed upon the subject) not dependent on the particular sensory channel used to deliver task-relevant information.

The feedback effects of sex steroid hormones on pituitary gonadotropin release in Turner's syndrome and Klinefelter's syndrome.

The present study was designed to elucidate the feedback relationship between the release of pituitary gonadotropins and sex steroid hormones in Turner's syndrome and Klinefelter's syndrome. LH-RH stimulation test was employed to evaluate the effects of sex steroids on the release of gonadotropins. The release of gonadotropins in response to LH-RH as well as in baseline level was suppressed after the treatment with estrogen (mestranol 0.08 mg/day) for 10 days, followed by the treatment of the same period with estrogen (mestranol 0.08 mg/day) and progesterone (chlormadinone acetate 2.0 mg/day) in combination in both syndromes. The inhibitory effect of the combined treatment was greater than that of the treatment with estrogen alone. Administration of testosterone propionate (25 mg/day) for 3 days resulted in suppression of the release of both gonadotropins in baseline level and in response to LH-RH in both syndromes, but the suppressive effect appeared to be less complete as compared with that of estrogen or estrogen-progesterone. It was thus verified that the feedback interaction between the pituitary gonadotropin release and sex steroids such as estrogen, estrogen-progesterone or testosterone was operative in the same fashion in the patients with Turner's syndrome and Klinefelter's syndrome.

Alpha subunit of glycoprotein hormone: presence in peripheral serum after LHRH.

A method is described for the selective measurement of human luteinizing hormone (hLH) and alpha subunit. The assays employ highly purified tracers of hLH and of subunit of human chorionic gonadotropin (hCGalpha) and a "mixed population" of antibody: Population I (directed against determinants on beta subunit of hCG) and Population II (directed against determinants on alpha subunit of hCG). The former is present in greater concentration than the latter. When the mixed antibody is used at higher dilution (1:1.2x10(6)), Population II is effectively diluted out, and using 125I-labelled hLH as tracer, the assay recognize hLH, hCG and their beta subunits 20-50 times more sensitively than hCGalpha. When the mixed antibody is used at fivefold higher concentration, Population I is present in relative excess and acts as a "sink" for hCG, hLH and their beta subunits. Under these conditions, using 125I-hCGalpha as tracer, the assay recognize the alpha subunit 20 to 50-fold more sensitively than hLH and hCG. These assays have been employed in the study of sera which have been filtered over Sephadex G-100. Alpha subunit was detected in serum within minutes after intravenous injection of luteinizing hormone releasing hormone (LHRH) in four subjects tested.

Preliminary characterization of a major allergen of timothy grass pollen.

Previous studies with timothy pollen extracts demonstrated that a low molecular weight dialyzable fraction, antigen D, possessed the allergenic determinant of the major allergen of timothy pollen (allergen B). Bio-gel P-2 gel-filtration of the dialysate fraction resulted in the isolation of a fragment, antigen D3 with a molecular weight near 1000. The relationship of antigen D3 to the antigenic and allergenic determinants of allergen B was evaluated by passive cutaneous anaphylaxis (PCA) in guinea pigs sensitized with rabbit antitimothy sera and by allergen-induced histamine release from monkey lung tissue passively sensitized with the serum of timothy-sensitive patients. These studies demonstrated that 10 units of antigen D3 gave 1) a 70% inhibition of PCA reactions in guinea pigs challenged with allergen B, and 2) a 73% inhibition of allergen-induced histamine release from monkey lung tissue sensitized with timothy reagin and challenged with the crude pollen extract (WST). Although chemical characterization studies of the antigen D3 fragment are still being carried out it appears that the major components fo this fragment are 1) a flavonoid pigment - quercitin, 2) a disaccharide moiety - cellobiose, and 3) the amino acid threonine linked together in an )O-glycosidic type linkage.

Formal genetics of the HL-A region.

The extreme polymorphism of the HL-A system is due to the presence of two (SD1, SD2) and perhaps three linked polyallelic genes. The distinction of "bridging antibodies" (reacting with several HL-A specificities recognizing separate sites on the HL-A molecule) from the main HL-A determinant as it is demonstrated by absorption/inhibition experiments increases this complexity. The HL-A linkage group is composed of other systems: LD1, LD2, PGM3, ADA (?), P, ME1, IPO-B and possibly a "hay fever gene". No gametic or zygotic selection was found in spite of the presence of HL-A antigens on spermatozoa. Mixed lymphocyte reaction (MLR) is principally governed by LD genes. The main (LD2) gene is probably situated outside the interval SD1, SD2, near SD2. Other LD genes (LD1 inside the interval SD1-SD2 and LD3) are suspected. The presence of an immune response gene (Ir) has not yet been demonstrated although several diseases associated with specific SD2 antigens are known. These different genes (SD1, SD2, LD1, LD2, LD3 and Ir) probably form a functional unit in the allo-immunozation.

[Cat-eye syndrome. Clinical and cytogenetical differentialdiagnosis (author's transl)].

We report a 5 1/2-year-old girl whose clinical symptoms are consistent with diagnosis of the cat-eye syndrome. The prominent symptoms are: anal stenosis, preauricular tags and pits, coloboma of the iris, doubling of the pelvis and ureter on both sides, vesicourethral reflux on the right side and normal mental development. Leucocyte alkaline phosphatase is normal. Chromosomal analysis shows a supernumerary submetacentric chromosome. This extra chromosome is smaller than the G-group chromosomes and has satellites on the short and long arms. Autoradiography after 3H-thymidine incorporation shows a late-labeling marker chromosome. After using the Giemsa-banding technique, the chromatides demonstrate dark bandings with only soft, unstained satellites. With the fluorescence method, one can see spotlike fluorescence of the satellites on both arms and diffuse fluorescence of the hetero-chromatic segments. In addition, the C-bandings demonstrate a homogeneous dark staining of the chromatids, but we did not find stained satellites. Using the Giemsa-11 technique one can see the 47th chromosome with predominantly heterochromatic parts, but small euchromatic segments are visible between them. Satellites are unstained. Using currently accepted cytogenetical methods, it is not possible to identify the origin of this supernumerary marker chromosome.

The antigenic interrelations of some mammalian IgG subclasses detected with cross-reacting fowl antisera to human and mouse IgG-Fc.

Two fowl antisera to plyclonal human IgG and one to a mouse IgG1 myeloma protein, after absorption either with homologous F(ab)2 alone, or also with myeloma proteins of known subclasses, cross-reacted in immunoelectrophoresis or immunodiffision withthe IgG of many mammalian species. Often more than one IgG precipitation line was formed in the corss-reacting systems. By testing isolated IgG1 and IgG2 fractions from mouse, rat, guinea-pig, bovine and dog sera, it was shown that the distinct arcs seen with whole serum corresponded to the known IgG subclasses of these species. With rabbit serum as antigen, the two anti-human-Fc sera diffusing from neighboring wells formed a 'spur', showing that they were reacting with determinants on different molecules, and that therefore rabbit IgG contains two antigenically distinct Fcpopulations. With the possible exception of canine IgG1 and human IgG4, no antigenic homologies were found between the subclasses of different species, thus supportingthe view obtained from sequence data, namely that IgG subclasses within species havearisen independently.

Comparison of lymphocyte populations bearing surface immunoglobulins in avian bone marrow, bursa, spleen and thymus.

Rabbit anti-chicken gamma-globulin was labeled with 125I and then incubated with cells from the bursa, thymus, spleen, and bone marrow of 4- and 8-week old birds. The same procedure was carried out on 11-week-old agammaglobulinemic chickens. Autoradiography revealed that the majority of large, medium, and small bursal lymphocytes bind the antibodies while labeled lymphocytes of each type in the spleen and thymus never exceeded 11 or 4 percent, respectively. Labeled medium and small lymphocytes in the bone marrow increased from 4.2 and 1.7%, respectively, at 4 weeks of age, to 9.5 and 8.8%, respectively, at 8 weeks of age. Labeled lymphocytes of all sizes were completely absent in all tissues of agammaglobulinemic chicks, including the marrow. Therefore, the increase in frequency of labeled lymphocytes in the bone marrow with age may be a result of recruitment of cells from the bursa of Fabricius. The majority of lymphocytes in the bone marrow do not label. Therefore, lymphocytes from the bone marrow may be T cells, subsets of B cells, or neither T or B cells.

Natural cytotoxic reactivity of mouse lymphoid cells against syngeneic acid allogeneic tumors. I. Distribution of reactivity and specificity.

Lymphoid cells from many normal mice of a variety of inbred strains were found to have reactivity, in a 51Cr release cytotoxicity assay, against several syngeneic and allogeneic tumors. Very high reactivity was seen with effector cells from athymic nude mice, which was consistent with other evidence that the reactivity was not T-cell dependent. Target cells susceptible to lysis included tumors induced by oncogenic type-C viruses but also tumors induced by other means and expressing endogenous type-C viruses. The levels of natural reactivity were influenced by age, with highest cytotoxicity produced by cells from 5- to 8-week-old mice. Lymph-node cells, spleen cells, peritoneal exudate cells and peripheral blood lymphocytes all had cytotoxic reactivity. The specificity of the reactions was analyzed in detail by ana inhibition assay. Evidence was obtained for natural reactivty against several different antigens, each apparently associated with expression of murine endogenous type-C viruses.

Residues in broiler chick tissues from low level feedings of seven chlorinated hydrocarbon insecticides.

Two chlorinated insecticide feeding studies from 1967-1968, using broiler chicks, have been completed. In Study I, lindane, heptachlor epoxide, dieldrin, endrin, methoxychlor, and DDT were fed in combination at 3 levels: 0.05, 0.15, and 0.45 ppm. Data show that, in fat tissues, heptachlor epoxide attained a level approximately 20 times the respective levels in the feed; dieldrin 15 times; endrin 10 times; p,p'-DDT 9 times; lindane 3 times; and o.p'-DDT less than the feeding levels. Of the DDT metabolites, p.p'-DDE and p,p'-DDD, only p,p'-DDE was significant at 3 times the 0.45 ppm feeding level. Endrin ketone, a metabolite of endrin, reached plateau levels approximately equal to feeding levels. All residue levels in liver tissues were less than 0.02 ppm. In Study II, technical chlordane was fed singly at the 0.05, 0.15, and 0.45 ppm levels. Results are tabulated for both total chlordane and for 6 identifiable isomers. Total chlordane in fat tissues attained plateau levels 3-5 times the respective feeding levels. Total chlordane levels in liver and breast tissues were all less than 0.01 ppm.

Vitamin K and the biosynthesis of prothrombin. V. Gamma-carboxyglutamic acids, the vitamin K-dependent structures in prothrombin.

Tryptic peptides obtained from normal prothrombin have been compared with those obtained from prothrombin synthesized by cattle given the vitamin K antagonist dicumarol. Two peptides were found which contain vitamin K-dependent structures. These peptides contain residues 4 through 10 and residues 12 through 44, respectively. One of these (residues 4 through 10) has previously been shown to contain gamma-carboxyglutamic acid residues. Digestion of this peptide with aminopeptidase M and carboxypeptidase B yielded a tetrapeptide (residues 6 through 9). Mass spectra of this peptide showed that it has the structure Leu-Glu(CO2)-Glu(CO2)-Val. The structure of the peptide containing residues 12 through 44 was determined by automated degradation in a peptide sequenator. The modified glutamic acid residues were identified by mass spectrometric comparison with the thiohydantoin derivatives of synthetic gamma-carboxyglutamic acid. This approach unequivocally demonstrated that all of the first 10 glutamic acid residues in prothrombin are carboxylated to form gamma-carboxyglutamic acid residues. Evidence is also presented that indicates that these gamma-carboxyglutamic acid residues constitute the entire vitamin K-dependent modification of prothrombin.

Translaminar growth of axons in the kitten dorsal lateral geniculate nucleus following removal of one eye.

The distribution of retinal afferents to the dorsal lateral geniculate nucleus of the cat following the early removal of one eye has been studied using autoradiographic as well as degeneration techniques. The degeneration techniques have confirmed earlier observations that axons from the remaining, normal eye grow across laminar borders into the deafferented lamina A. Autoradiographic techniques, however, have shown further examples of translaminar growth between lamina A nadlamina A1 as well as between lamina A1 and lamina C. Although the possibility exists that such growth also occurs between the other C laminae (C1, C2 and C3) such growth is harder to demonstrate since no well defined interlaminar regions exist in this part of the nucleus. In the monocular segment of the nucleus degeneration techniques have shown some translaminar invasion of the deafferented lamina A. Autoradiographic techniques have demonstrated similar growth as well as an abnormal invasion of the most lateral parts of the monocular segment. This latter growth originates in the optic tract as it curves around the nucleus on the way to the superior colliculus.

Demonstration of a pallido-nigral projection innervating dopaminergic neurons.

Afferents to the substantia nigra from the neostriatum and globus pallidus were studied in the rat by means of the autoradiographic tracing technique. 3H-leucine was injected stereotaxically into either the globus pallidus or neostriatum. Twenty-four hours later the axoplasmic transport of labelled proteins to the substantia nigra was studied by light and electron microscopic autoradiography. In animals used for electron microscopy, degeneration of dopaminergic cells in the substantia nigra was induced by intraventricular injection of 6-hydroxydopamine 72 hours before sacrifice. After neostriatal injections, light microscopic analysis revealed heavy labelling of the globus pallidus and entopeduncular nucleus, but only background labelling of the subthalamic nucleus. There was preferential labelling of the zone reticulata of the substantia nigra, with significantly less labelling of the zone compacta. After pallidal injections, light microscopic analysis showed very light labelling of those parts of the caudate-putamen in the vicinity of the injection site. There was intense labelling of the subthalamic nucleus and heavy labelling of the entopeduncular nucleus. The zona compacta of the substantia nigra was also heavily labelled. There was considerably less labelling of the zona reticulata. The electron microscopic analyses showed that after neostriatal injections, autoradiographic grains in the substantia nigra were located preferentially over boutons which terminated on normal dendritic processes. After pallidal injections, however, grains were preferentially located over boutons synapsing with degenerating dendritic processes. The degeneration produced in these dopaminergic processes by 6-hydroxydopamine was invariably of the dark type. Except for the different association with degenerating vs. non-degenerating dendrites, the subcellular distribution of autoradiographic grains in the substantia nigra was the same after injection into either the globus pallidue or caudate-putamen. Approximately 80 percent of the grains were over axons or boutons which invariably made symmetrical synaptic contacts. These observations demonstrate the existence of a pallido-nigral projection which terminates preferentially on dopaminergic cells in the pars compacta of the substantia nigra. They also confirm previous studies indicating that the strionigral projection terminates mainly in the pars reticulata. These terminations appear to be principally to non-dopaminergic cells.

Radioimmunoassay of human serum antibody specific for adenovirus type 5-purified fiber.

A radioimmunoassay (RIA), utilizing a second antibody to separate immune complexes, was developed to provide a sensitive and specific measure of serum antibody to adenovirus type 5 (Ad 5) fiber. Purity of fiber antigen was ascertained by sodium dodecyl sulfate urea-polyacrylamide gel electrophoresis and isoelectric focusing in ampholyte pH gradients. After labeling with 125I to high specific activity, the iodinated fiber did not exhibit loss of antigenic reactivity and remained stable for 3 weeks when stored at minus 20 degrees C with supplemental protein. Rabbit anti-Ad 5 serum with a neutralization titer of 1:320 precipitated 50% of the labeled fiber at a serum dilution of 1:50,000 when tested by the RIA. In competition assays as little as 0.5 ng of unlabeled fiber per millimeter was sufficient to inhibit the 125I fiber-antibody reaction. Serum specimens from 20 volunteers, obtained before and after vaccination with purified Ad 5 fiber or hexon subunit vaccine, were tested by RIA, hemagglutination-inhibition (HI), and neutralization tests. A comparison of mean antibody titers of post-inoculation sera showed that the RIA was 300 and 1000 times more sensitive than the HI and neutralization tests, respectively. Moreover, 19 of the men who were negative by the standard serologic tests before vaccination were shown to have anti-fiber antibody, with a mean RIA titer of 1:1028. Specificity of the RIA was demonstrated by the lack of an increase in antibody to Ad 5 fiber among those individuals vaccinated with the hexon subunit. Thus, the development of a highly sensitive and reproducible RIA allows for the detection of antibody specific for the Ad 5 fiber in serum which contains antibodies to the different virion antigenic determinants associated with Ad 5.

Early steps in specific tumor cell lysis by sensitized mouse T lymphocytes. I. Resolution and characterization.

Addition of high molecular weight dextran to culture medium prevents the initiation of T lymphocyte-mediated killing by holding the cytolytic T lymphocytes (CTL) and target cells in suspension and preventing intercellular contact. Suspension in 10% dextran was used to interrupt the ongoing formation of adhesions between CTL and target cells already in contact in a centrifuged pellet. The results demonstrate that 1) firm adhesions form between CTL and target cells within 1 min at 37 degrees C; 2) once formed, these adhesions are stable at low temperature and are resistant to mechanical shearing forces; 3) these adhesions can be disrupted by EDTA; 4) immediately after the adhesions form, separation of the CTL from the target cells prevents lysis of the latter; 5) after incubation of targets adhering to CTL for an additional 6 min at 37 degrees C, removal of the CTL no longer prevents target cell lysis. Thus, target cells become "programmed" for subsequent lysis within a few minutes after contact with CTL, after which lysis occurs during the next several hours without further participation of the effector cell. At 15 degrees C, adhesions form 1/17 as fast as at 37 degrees C. Programming of target cells for lysis occurs 1/76 as fast at 15 degrees C as at 37 degrees C. Thus, the programming for lysis step is about 4-fold more temperature dependent than the adhesion step. In addition to being detected by subsequent target cell lysis in 10% dextran, the adhering cell clusters can be counted with low power microscopy. This permitted verification that EDTA separates the clusters after programming for lysis is complete. Moreover, the great majority of the clusters seen at 37 degrees C are antigen-specific. Knowledge of the cluster size distribution and the subsequent level of lysis permits the deduction that not less than 6% of the sensitized peritoneal cell populations used were CTL.

Normal tissue alloantigens and genetic control of susceptibility to tumors: microcytotoxicity studies on resistant C3Hf and susceptible (A X C3Hf) F1 mice inoculated with transplacentally induced C3Hf lung tumor.

The immune response of mice to a transplacentally induced lung tumor was investigated with the microcytotoxicity (MC) assay. The tumor, originally induced in C3Hf mice, does not grow readily when transplanted to normal syngeneic C3Hf recipients. It grows readily, however, in (A C3Hf)F1 hybrids and in strain C3H mice, which express in their normal lung tissue a component which constitutes a strong lung tumor-associated transplantation antigen (TATA) in C3Hf mice. Both lung tumor-immunized C3Hf and tumor-bearing (A X C3Hf)F1 and C3H mice possessed lymphoid cells reactive against cultured lung tumor cells in the MC assay. Reactivity was also observed against cells cultured from normal lungs of (A X C3Hf)F1 and C3H mice, but not against cells similarly cultured from C3Hf of C57BL/6 mice. Anti-tumor MC was inhibited by serum-blocking factors present in some but not all tumor-bearing and tumor-immunized mice. The MC assay and detection by it of serum-blocking factors does not distinguish the effective anti-C3Hf lung tumor immune response of immunized C3Hf mice from the ineffective immune response of tumor-bearing (A X C3Hf)F1 and C3H mice. Furthermore, in lung tumor-bearing mice cells reactive in the MC assay may be directed against a normal tissue antigen rather than a tumor-associated antigen.

Immunologic effects of morphine administration in rabbits.

Long-term effects of morphine administration or immunologic test responses were studied in female rabbits. Implantation of morphine-containing pellets was found to be more effective than injection of morphine sulfate solutions in promoting increased serum binding of 140-morphine. A large part of the increased morphine binding by sera associated with administration of morphine was found in serum fractions containing gamma-globulin and was absent in gamma-globulin-free fractions. These sera showed some degree of specificity for the morphine configuration in tests with other narcotics. They also gave positive immunologic test reactions in passive hemagglutination and radial immunodiffusion tests involving serum albumins conjugated with morphine derivatives. Other evidence for immunologic responsiveness against morphine by morphine-pretreated rabbits was shown by cutaneous hypersensitivity reactions against morphine-carrier conjugates and by a diminution of the serum morphine-binding response in rabbits given an immunosuppressive dose of cyclophosphamide. Failure of naloxone, a morphine antagonist, to alter the serum morphine-binding response suggested that serum levels of the morphine-binding globulin studied here were not direclty related to morphine withdrawal.

Desmosomes, filaments, and keratohyaline granules: their role in the stabilization and keratinization of the epidermis.

Components of desmosomes, filaments, and keratohyaline granules were studied by electron microscope and biochemical methods to clarify their role in the stabilization and keratinization of the epidermis. Isolated desmosomes are composed of 76% protein, 17% carbohydrate, and 10% lipid. The bulk of protein consists of a "spectrin"-like fibrous protein, presumably present in the plaque, and of glycoproteins in the desmosomal interspace. The main component of filaments, prekeratin, is a low-sulfur alpha-protein composed of a pair of three-chain subunits with non-alpha-helical segments separated by 200 A-long alpha-helical regions. The major component of isolated keratohyaline granules, the amorphous particulate material, is formed by a high-sulfur protein with a single-type of polypeptide chain. Polypeptide chains comparable to those found in prekeratin and keratohyaline granules were recovered from extracts of horny cells. Within the living part of the epidermis, filaments hypothetically form a cytoskeletal system which is anchored to desmosomes by a filamentous plaque protein. Glycoproteins are involved in the formation of strong junctions between the cells which enable the living part of the epidermis to respond as a whole to mechanical stress. The stratum corneum is stabilized by a similar system in a consolidated state which is less extensible. Horny cells are enveloped by a thickened membrane and the interfilamentous spaces are filled with various proteins including the sulfur-rich amorphous protein found in keratohyaline granules.

Microfilaments in the external surface layer of the early amphibian embryo.

A comparison was made by transmission electron microscopy of the microfilaments in the surface layers of the early embryos of Triturus alpestris and Xenopus laevis at stages of development up to neurulation. Actin-like filaments which bound heavy meromyosin (HMM) were found in cell extracts of all stages, but were comparatively rare in the newly fertilized egg. Ten nm microfilaments were present throughout development in Xenopus, and from the mid-neurula stage in Triturus. Both kinds of microfilament were located in the circumferences of superficial ectoderm cells at the level of the apical junctions, the 10 nm microfilaments in association with desmosomes which began to develop before gastrulation in Xenopus. The accumulations of microfilaments in the apical constrictions, which form in ectoderm cells of Triturus early gastrulae when dissociated in a calcium-free medium, suggest that they are contractile elements. In the absence of such accumulations in the cell apices, the reverse curling exhibited by Xenopus ectodermal explants is attributed rather to a separation of the cells' lateral borders. Cytochalasin B (5 mug/ml) caused ectodermal explants from the early gastrulae of both species to disaggregate. With the rupture of the apical junctions there was a disorganization of the associated microfilamentous layer.

Cross-resistant relationships among the aminoglucoside antibiotics in Mycobacterium tuberculosis.

Phenotypes of isolates of Mycobacterium tuberculosis H37RV showing resistance to the aminoglucoside antibiotics streptomycin, viomycin, kanamycin, capreomycin, tuberactinomycin N, lividomycin and paromomycin could be grouped into the following types: (I) resistant only to different levels of streptomycins; (2) resistant only to a low level of kanamycin; (3) triply resistant, to low levels of viomycin, tuberactinomycin N and capreomycin; (4) triply resistant, to a low level of kanamycin and high levels of lividomycin and paromomycin; (5) quadruply resistant, to a low level of capreomycin and high levels of kanamycin, lividomycin and paromomycin; (6) hextuply resistant, to high levels of viomycin, tuberactinomycin N, capreomycin, kanamycin, lividomycin, and paromomycin. Three modificatied types of the latter were also observed. Appearance rates of the six types were estimated as 10(-6) to 10(-9), 10(-6), 10(-6) to 10(-7), 10(-8), 10(-8), and 10(-8) to 10(-9), respectively, in a total viable population of the parent strain. Mutations to all phenotypes were considered to be produced by single mutations. According to cross-resistance relationships, aminoglucoside antibiotics were classified into three groups: (I) streptomycin; (II) viomycin, tuberactinomycin N and capreomycin; (III) kanamycin, lividomycin and paromomycin. No cross-resistance relationship between streptomycin and other antibiotics was observed. Resistances to viomycin, tuberactinomycin N and capreomycin occurred by single mutation to type 3. Resistances to kanamycin, lividomycin and paromomycin occurred by single mutations to types 4 and 5. Low resistance to capreomycin was produced by mutation to type 5. Therefore capreomycin was considered to be an intermediate between the second and third groups. These two groups had a close relationship, as resistance to all six agents in these groups could be produced by a single mutation to type 6 (and its modified types).

On the effect of brain phospholipase A1 on specifically labelled glycerophospholipids in the course of subacute sclerosing panencephalitis.

The effect of phospholipase A1 of human brain on 1,2-diacyl-sn-glycero-3-phosphorylcholine, -ethanolamine and -serine, specifically labelled with different fatty acids at either the 1 or 2 position, was determined in subacute sclerosing panencephalitis. An increase of approximately 40% in the specific activity of phospholipase A1 could be observed for all substrates investigated during the demyelinating disorder. On investigating the specific activity of the enzyme with various molecular species of phosphatidylcholine and -ethanolamine, labelled at the 1 position with different radioactive fatty acids, we found that the phospholipase A1 preferentially removed those fatty acids from the 1 position of phosphatidylcholines that have the fewest double bonds, while oleic and linoleic acid were released at almost similar rates from phosphatidylethanolamine.

The effects of ganglionic blockade, reserpine and vinblastine on plasma catecholamines and dopamine-beta-hydroxylase in the rat.

In rats, chronic ganglionic blockade induced by repeated doses of chlorisodamine rapidly and profoundly lowered plasma norepinephrine, but plasma dopamine-beta-hydroxylase (DBH) activity, even after 5 days treatment, was not significantly reduced. Long-term chlorisondamine treatment did not alter cardiac DBH or rapid axonal transport of DBH in sciatic nerve. Blockade of alpha adrenergic receptors by administration of repeated doses of phenoxybenzamine resulted in elevated levels of plasma catecholamines, but produced no change in plasma DBH. Chronic reserpine treatment (2.5 mg/kg on alternate days) increased plasma DBH after 2 and 5 days, whereas vinblastine (3 mg/kg) caused a progressive fall in enzyme activity in plasma over the same time period. It is concluded that plasma DBH activity does not closely parallel adrenergic function and neurotransmitter release in the rat. The level of DBH in plasma appears to reflect the rate of enzyme synthesis and axonal transport, It is likely that mechanisms other than stimulation-coupled exocytotic release determine levels of DBH activity in plasma.

[Ultrastructure of Sarcocystis tenella. I. Endozoïte (after negative staining)].

The employment of negative staining technics for the endozoites (cyst stages) of Sarcocystis tenella allowed the elucidation of certain aspects of their fine structure. The conoid consists of similar to 20 oblique fibers and is surmounted by a ring with regular ornamentation. In the conoid's interior there are 2 excentric parallel microtubules which extend posteriorly for a considerable distance into the adjacent cytoplasm. The fibers of the conoid, intraconoid microtubules, appear to have the same diameter and structure as the 22 subpellicular microtubules. They are "cemented" anteriorly into a periconoidal ring which surrounds the conoid. The "reticulated" pellicle has certain differentiations: the micropore, surrounded by a "fibrillar" element, similar to 10 subcircular structures arranged into an anterior crown, and 11 rows of granules converging toward the posterior end. The sarconemes look like rice grains which, contrary to previous statements, are independent of one another. It is established that there are only 2 rhoptries.

Serologic abnormalities in spontaneous and drug-induced systemic lupus erythematosus.

Extensive serologic changes occur in systemic lupus erythematosus (SLE) which are probably secondary to unknow primary cause(s). The New Zealand hybrid mouse model most likely has a viral-induced disease and does not show many of the clinical features of the human disease. The best example of human SLE which provides a clue to etiology is the drug-induced type, particularly that due to procainamide. In these patients it is possible to study the development of serologic changes prior to the onset of clinical manifestations, and then observe regression of the clinical and serological changes on withdrawal of the medication. Although there is a rough correlation between the many serologic abnormalities and the clinical picture, enough exceptions exist, so that single tests such as serum complement, anti-DNA antibodies, per cent DNA binding, and others, cannot be used as a sine qua non for management. Care of the patient still remains a clinical problem guided by various laboratory procedures but not dependent on any one. Alkyating agents have a limited role in the treatment of lupus nephropathy and cutaneous vasculitis but azathioprine is probably of no value in SLE.

Influence of oestrogen on experimental pyelonephritis caused by Escherichia coli.

Strains of Escherichia coli isolated from patients with suspected pyelonephritis had a strong predilection for growth in kidney tissue. Viable bacterial counts in experimentally infected mice showed that after 96 hours approximately 98% of the total-body count was found in the kidney. Strains of E. coli isolated from cases of gastroenteritis in man and animals had little or no tendency to grow in the kidney. Treatment of male and female mice with oestrogen significantly enhanced the growth of "pyelonephritic" strains in the kidney, but had no effect of any kind on the growth of "gastroenteritis" strains. These preliminary results suggest that oestrogen may predispose to the development of kidney infection in the female and that there is an important link with the virulence of the E. coli concerned. Only those strains that have a natural predilection for growth in the kidney are likely to be influenced by oestrogen.

Retroperitoneal fibrosis during treatment with methydopa.

Retroperitoneeal fibrosis (R.P.F.) and a positive direct Coombs test developed in a patient who had received 1.4 kg. of alpha-methyldopa over a period of 5 years. Immunofluorescence microscopy demonstrated deposits of IgG, IgM, and IgA on the collagen fibres of the R.P.F. Biopsy speciments of the temporal artery, the right kidney, and the R.P.F. did not show signs of general arterial disease on light, immunofluorescence, and electron microscopy. Drugs which provoke autoimmunisation and interfere with nervous transmission are known to induce deposition of collagen or R.P.F. This suggests that R.P.F. in this patient was probably caused by alpha-methydopa.

Antigenic relationships between Candida albicans and various yeasts as reflected by immunoglobulin-class specificity.

A group of guinea pigs was inoculated into the foot pads with a single dose of Candida albicans in complete Freund's adjuvant, while another group was similarly inoculated once in the foot pads but also several times intramuscularly, with Candida alone. All guinea pigs were bled at different intervals after immunization and sera were separated chromatographically into IgG and IgM fractions. In order to study the antigenic relationships as reflected by immunoglobulin-class specificity, IgG and IgM fractions and whole sera obtained from guinea pigs differently immunized, were tested for the presence of agglutinins against C. albicans, six other species of Candida, and species of the ascosporogenous genera Saccharomyces, Kluyveromyces and Schizosaccharomyces. The results show that (1) only IgG fractions of the different sera prepared contained the specific anti-C. albicans antibodies; (2) IgG and IgM fractions of the sera obtained from a single inoculation did not reveal a specific pattern expressing antigenic relationships of the yeast studied, and (3) the IgM fractions of the sera obtained from several inoculations had a more homogenous pattern of reactivity, since mainly these contained the agglutinins against the ascosporogenous yeast species.

Hepatic complications of antituberculous therapy.

Drug hepatitis occurred in 0-32 per cent of 7492 patients receiving antituberculous therapy, while the overall incidence of drug reactions was estimated at 9 per cent. PAS was the most common cause of drug hepatitis among the 38 patients analysed. The clinical, biochemical and haematological picture of antituberculous drug hepatitis was found to be fairly uniform. However, the patients with definite PAS hepatitis had lower SGOT values than those in whom there was uncertainty whether PAS or INH was implicated. Premonitory symptoms were present in all but four patients before the onset of jaundice. One or more of the features associated with dry hypersensitivity reactions, such as fever, rashes, lymphadenopathy, arthralgia, leucocytosis, eosinophilia and atypical monocytes were present in 89 per cent of cases so that confusion with viral hepatitis seldom arose. Sensitization time was less than three months in all except three patients, who were considered to be suffering from viral hepatitis. While no patients with PAS hepatitis died, the overall mortality was 17 per cent. A review of the literature stresses the frequency of asymptomatic elevations of SGOT, the value of clinical surveillance during the early months of therapy and the importance of stopping all therapy immediately warning symptoms appear.

Clinical aspects of invasive carcinoid tumors.

In a retrospective study at the University of Louisville Affiliated Hospitals, 42 patients with carcinoid tumors not arising in the anorectal area were identified in ten years (1962-1972). The ileum was the organ most frequently involved with primary tumor (28%). The nonappendiceal gastrointestinal tumors were multiple in 28%, metastatic in 66%, and associated with a second malignancy in 25%. Of the symptomatic small-bowel tumors, 83% were metastatic at the time of diagnosis. Carcinoid syndrome was observed in only two patients, both of whom had liver metastases and elevated urinary 5-HIAA levels. Resections for cure were done on 25 patients, palliative resections on six, and biopsy on six. Six tumors were from autopsy meterial. Among the 24 patients treated and followed up for five years, the survival rate was only 16%. In those patients having resection for cure, the five-year survival rate was 39%, exculding appendiceal tumors. The advanced stage of disease at time of discovery and the dismal prognosis for invasive carcinoids are contrary to many clinicians' impressions of the nature of carcinoid tumors but entirely consistent with several other recent reports (James Ewing Society meeting, April 1973).

Direct cell mediated lympholysis. A test of allograft-rejection in human kidney recipients.

In kidney transplanted patients a clear coincidence was observed between clinical signs of allograft rejection and the presence in the peripheral blood of killer cells able to lyse either PHA lymphoblasts from the actual donor or selected unrelated individuals. In recipients with non-immunological complications such as leakage on the graft ureter or primary anuria caused by renal ischaemia, no cellular cytotoxicity against specific or selected target cells was observed. The specificity of this Cell Mediated Lympholysis in two of the cases reported could not be explained by the serologically detectable HL-A antigens, indicating the existence of other determinants of importance for the killing capability of in vivo produced effector cells.

Ir-associated murine alloantigens. Serological and chemical definition of Ia specificities associated with the H-2-b haplotype.

Strains bearing recombinant H-2 haplotypes with known positions of recombination have been used to demonstrate antibodies reacting with Ia antigens associated with the H-2-b haplotype in several H-2 antisera. Two new Ia specificities, Ia.8 and Ia.9, both determined by genes in the Ir-1A subregion, have thus been defined, and the strain distribution of these specificities has been determined. Immunoprecipitation of radio-labeled membrane antigens has been used to characterize Ia.9 and to distinguish it chemically from Ia.7 in a membrane preparation from a strain possessing both specificities.

Argyrophil cell carcinomas (apudomas) of the uterine cervix. Light and electron microscopic observations of 5 cases.

Of a series of 97 invasive carcinomas of the cervix, 5 were found to have argyrophil tumor cells, and 3 of these 5 tumors were studied by electron microscopy. The ages of the 5 patients ranged from 36 to 49 years, with a mean age of 42.4 years. The morphologic features of these five tumors were well consistent with those described on a variety of endocrine polypeptide neoplasms such as thyroid medullary carcinomas, carcinoids, pancreatic islet-cell tumors, and oat cell carcinomas of the lung. Microscopically, the 5 tumors were characterized by the formation of solid-sheets, ribbons, streams, and rosettes. They were characterized electron microscopically by the presence of neurosecretory-type granules, the abundance of intracytoplasmic microfilaments, the absence of tonofibrils, and the paucity of desmosomal attachments. On the basis of the microscopic, electron microscopic and cytochemical characteristics, it is suggested that the tumors are a specific type of cervical carcinoma derived from the argyrophil cells, normally found among the linings of the endocervical glands and the cervical squamous epithelium. We believe these 5 tumors should be regarded as an endocrine tumor, another member of apudomas.

[Immunochemical study of the antigenic structure of bacteria of genus Bordetella. IV. Fractionation of B. pertussis extracts and study of the immunochemical and biological properties of the isolated fractions].

Fractionation of an extract of pertussis microbes was carried out with the aid of gel-filtration on Sephadex G-100, ion-exchange chromatography on DEAE-cellulose, and preparative electrophoresis. Fractions differing in serological, immunogenic activity and the content of antigenic components were isolated. In using the method of gel-filtration of sefadex G-100 the greatest serological, immunogenic and histamine-sensitizing activity was possessed by the high-molecular fraction containing 8 of 11 antigenic components detected in the initial extract. The antigenic components were distributed into 5 fractions by the ion exchange chromatography on DEAE-cellulose. The greatest serological activity was possessed by fractions exiting from the column at the 0.01--0.04M interval of the phosphate buffer concentration. A method of preparative electrophoresis from the pertussis microbes extract was applied and two fractions were isolated from the anode and the cathode zones, each containing 4 antigenic components only, but possessing serological and immunogenic activity and having no histamine-sensitizing properties.

The ultrastructure of the accessory sex organs of the male rat 10. Effect of chlormadinon.

The present paper describes the fine structure of the accessory sex organs of the male rat as seen after administration of the anti-androgenic compound chlormadinon acetate for 20 days. There was a general involution of the organs with macroscopic atrophy and reduced amount of secretory material. Ultrastructurally the cells contained less organelles as compared with the controls, loss of cytoplasm and reduction both of cell height and width. The major changes also included reduction of the Golgi areas and the rough endoplasmic reticulum. Within the dorsal lobe prominent nuclear changes were found. The alterations observed in the present study are similar to the changes which were found in rats treated with another anti-androgenic compound, Cyp A. It is concluded that these two antiandrogenic compounds exert their effects through similar mechanisms with a specific influence on the prostate and the seminal vesicles.

Experimental rheumatoid arthritis-like features induced by prolonged sensitization with focal antigens.

Prolonged sensitization with emulsion of an autologous or isologous subcutaneous abscess of Arthus type induced by injection of hen egg-white was performed in 34 rabbits which were divided into (high responder and intermediate responder groups (H- and M-groups) according to individual difference of immune responses. The development of a rheumatoid factor-like substance (RFLS) was demonstrated after 30 experimental days and subsequently observed in 21 out of 33 rabbits. There was no significant difference in the incidence of RFLS between both groups. As to the relation between the development of RFLS and types of focal antigens, the group of the autologous W-substance showed a higher incidence of RFLS than the N-substance. Acute and/or chronic synovitis was demonstrated in 13 of 33 rabbits and inflammatory changes were more intensive and extensive in the later period of experiment. Presence of RFLS in the affected synovial tissues, chiefly in the cytoplasm of plasma cells and mononuclear cells, occasionally in a free state was revealed by immunofluorescent study, and depositions positive for IgG and beta 1C were observed in the wall of blood vessels and fibrinous thrombi in the affected synovial tissues.

The genetic structure of a tribal population, the Yanomama Indians XI. Gene frequencies for 10 blood groups and the ABH-Le secretor traits in the Yanomama and their neighbors; the uniqueness of the tribe.

In this paper we present the results of blood group typings for a total of 33 villages distributed among five South American Indian tribes--Yanomama (21 villages), Makiritare (eight villages), Macushi (two villages), Piaroa (one village), and Wapishana (one village). These new results for the Yanomama and Makiritare tribes have been combined with those previously reported to allow a better appreciation of the distribution of allelic frequencies in the tribes. The relationship of the Yanomama to other South American Indian tribes is investigated using data on six polymorphic loci (Rh, MNS, Fy, Jk, Di, Hp). By use of four genetic measures (two of genetic relationship and two of genetic diversity), we demonstrate that the Yanomama are genetically unique among a sample of 20 South American tribes. In addition, the Yanomama show somewhat less genetic diversity for the six loci analyzed than the average South American tribe. Taken together, these results indicate a rather long period of isolation for the population antecedent to the Yanomama--perhaps since the time of entry of man into the South American continent. The pattern of genetic relationships and genetic diversity for the 20 tribes is consistent with the hypothesis that evolution in South America proceeded by a process of fission-fusion leading to isolation of subpopulations with subsequent genetic differentiation as a consequence of population isolation. The uniqueness of the Yanomama appears to stem entirely from such a process, there being no evidence of any selective differential for the loci analyzed.

Substructure in rod photoreceptor membranes.

Frog retinae, fixed only in buffered glutaraldehyde, were embedded for sectioning in glutaraldehyde polymerized with urea. In suitably thin sections globular substructures were seen in negative contrast after ionic staining with uranyl acetate and lead citrate, or after staining with neutralized phosphotungstic acid. Efforts to extract at least some of the lipid from sections before ionic staining enhanced the visualization of the "globules". Exposure to KMnO4 solution, used as an oxidative section stain, also outlined globular substructure in negative contrast, but with the additional feature that positively stained surface "leaflets" associated with the aqueous compartment were well defined. Staining sections with OSO4 vapor resulted in positively stained membranes, but without any evident substructure. However, when sections which previously had been exposed to OSO4 vapor were secondarily stained with uranyl acetate and/or lead citrate, positively stained globular substructures then were revealed. The globular substructures always were centered in the hydrophobic core region of the disc membranes, and symmetrically spanned the full thickness of this layer. The diameter of individual particles approximated 50-55 A. Reasons are presented for the supposition that the evident globules incorporate at least hydrophobic components of rhodopsin molecules. Findings are discussed in relation to various models of disc membrane organization that have been proposed in recent years.

Eustachian tube lymphatics.

Serous otitis media is the most common cause of hearing impairment. The role of lymphatic obstruction in the pathogeneis of serous otitis media is significant. A method for removeal of the human Eustachian tube specimen and two techniques for identification of Eustachian tube lymphatic capillaries are described. One involves the antemorten intratympanic installation of Berlin blue. The other utilizes electron microscopy. Lymphatic capillaries cannot be reliably differentiated from blood capillaries with the light microscope. With electron microscopy, lymphatic capillaries can be differentiated from blood capillaries by differences in the basement membrane. The lymphatic capillary has gaps in the basement membrane with large nuclei in the wall. A blood capillary has a continuous basement membrane and sometimes red blood cells be be identified in the lumen. Using these methods, Eustachian tube lymphatic capillaries in the human are described for the first time in this report.

[Degradation of 14C, 3H, and 36Cl-labelled gamma-hexachlorocyclohexane by anaerobic soil microorganisms (author's transl)].

Up to 90% of the gamma-Hexachlorocyclohexane (gamma-HCH) applied to an anaerobic mixed bacterial flora enriched from an arable soil were degraded within 4-5 days. Degradation resulted in a rapid release of chloride and in formation of chlorine-free volatile metabolites. CO2 formation from the molecule was not detected. Investigations with 14C/3H- and 36Cl/3H double-labelled gamma-HCH indicated that the release of Cl and H did not occur in the ratio of 1:1. More Cl than H was split off. The volatile compounds contained more 14C than 3H. Gas chromatographic studies also showed the rapid decrease of gamma-HCH and the formation of several metabolities. gamma-Pentachlorocyclohexene was nto detected. Increasing O2-contents in the gas phase of cultures resulted in decreases of the compound's degradation. Release of chloride and of volatile metabolites were observed with O2 contents in the gas phase up to 5%. alpha-HCH was also, but more slowly as with gamma-HCH, degraded by the anaerobic mixed flora. Chloride was released and volatile, chlorine-free metabolites were found.

[Effect of ajmaline and its therapeutically used derivatives N-propylajmaline and di-monochloracetylajmaline on the functional refractory period and contractility of guinea pig atrium and aconitine arrhythmia in the rat].

1. In the isolated left atrium of the guinea pig ajmaline and di-monochloracetylajmaline (DCAA) show almost the same activity concerning prolongation of the functional refractory period. 2. In contrast to this N-propylajmaline (NPA) is much more effective than ajmaline. 3. NPA as compared to ajmaline and DCAA, however, shows a considerably smaller difference between the concentrations prolonging refractory period (I) and those decreasing contractility (II) in the guinea pig atrium (EC25). The quotient from I and II is 0.4 with NPA, 1.2 with ajmaline and 1.6 with DCAA. 4. Differences in efficacy similar to those observed in the guinea pig atrium are also found in experimental cardiac arrhythmias in the intact animal. NPA is much more effective than ajmaline regarding inhibition of extrasystoles, ventricular tachycardia and ventricular flutter due to aconitine infusion in the rat. In this experimental model DCAA shows slightly less activity than ajmaline; this difference is statistically significant.

Studies on mouse Moloney virus induced tumours: II. Detection of p30 in the serum of mice with Moloney leukaemia by in vitro blocking of complement dependent antibody mediated cytotoxicity.

Sera from Balb/c mice bearing Moloney leukaemia block complement dependent antibody mediated cytotoxicity of an antiserum prepared in rats against syngeneic Moloney virus induced lymphomata when either spleen cells from mice bearing Moloney leukaemia (M) or an in vitro line of Moloney virus transformed cells (MSC) are used as targets. This antiserum has been shown to recognize p30, the major internal virion protein, as a cytotoxic target on these cells. Viral particles were identified by electron microscopic examination of pelleted material obtained from leukaemic sera after high speed centrifugation. However, removal of virus did not affect the capacity of the leukaemic sera to absorb cytotoxicity of rat ILR-3 for MSC targets, and only depressed somewhat its ability to absorb activity of the same antisera against M targets. Virus-free leukaemic sera also blocks complement dependent antibody mediated cytotoxicity of an antiserum prepared in goats against the gs3 determinant of p30. This indicates that the material in leukaemic sera responsible for the in vitro block of antibody mediated cytotoxicity was p30. A lesser degree of block was observed with sera obtained from normal Balb/c mice, but the nature of material responsible is as yet undefined.

Bleomycin combination chemotherapy in the management of testicular neoplasia.

Eighty-three patients with Stage II or Stage III germinal neoplasia of the testis and 7 patients with extragonadal primary tumors were treated with bleomycin plus vinblastine, or a five-drug program, bleomycin plus cyclophosphamide, vincristine, methotrexate, and 5-fluorouracil. Of the 70 Stage III patients, there were 53 responses (75%), 22 complete and 31 partial. The mean survival of the complete responders is 100+ weeks, with 3 dead. The mean survival of the partial responders and nonresponders is 38 weeks and 33 weeks, respectively. There is a highly significant difference between complete responders vs. partial and nonresponders (p less than 0.01). Thirteen patients with nonmeasurable disease (Stage II and Stage III postresectional status) but at great risk to develop widespread metastasis were treated prophylactically after conventional therapy. Nine continue in complete response to 36 months. The 7 extragonadal primary patients showed 4 partial responses, none complete. Major toxicity was myelosuppression and also bleomycin pneumonitis in 5 of the 90 evaluable patients.

Effects of nitrosocarbaryl on BALB/3T3 cells.

Carbaryl(N-methyl-1-naphthylcarbamate) and its nitrosated product, N-nitrosocarbaryl, were tested for their effects of BALB/3T3 (clone A31) cells in culture. Nitrosocarbaryl, but not carbaryl, caused transformation of the BALB/3T3 fibroblasts, but neither chemical induced the complete expression of endogenous murine leukemia virus. Transformed cells differed from the parental control cells by loss of contact inhibition, change in morphology, growth in soft agar, growth to higher saturation densities, and tumorigenicity in normal newborn and irradiated weanling mice and athymic (nude) mice. Transformed clones were found to be negative for expression of RNA tumor virus antigens, viral reverse transcriptase, and infectious virus. Thus, it appears that nitrosocarbaryl can transform BALB/3T3 cells to tumorigenic cells with altered biological properties but without complete activation of RNA tumor viruses in the transformed cells. Expression of viral antigen in the transformed cells was inducible by iododeoxyuridine, indicating that the endogenous viral genome was retained in an unexpressed state.

Partial characterization of C-type particles in a cell line (WR-9) derived from a rat epidermoid carcinoma of spontaneous origin.

A C-type virus continuously released from a cell line (WR-9) derived from a spontaneous epidermoid carcinoma was purified by means of large-scale tissue culture techniques and high-volume zonal centrifuges. With the use of relatively pure virus concentrates, partial characterization of the virus has been accomplished. Up to 60 liters of spent culture medium from relatively low virus-yielding cultures were processed at a time through the Model K ultracentrifuge in order to obtain quantities of virus sufficient for convenient Tween-ether extraction of the major polypeptide (30,000 daltons). This structural protein having group-specific reactivity was purified and isolated by isoelectric-focusing techniques. A UV absorption peak (A280) was found to be coincident with a major peak of radioacticity at pH 8.6, the isoelectric point (pI) for rat virus gs antigen previously reported by other investigators. Because species-specific (gs-1) and cross-reactive (gs-3) determinants coexist on this protein, fractions containing the group-specific antigen were identified on the basis of the mammalian interspecies determinant (gs-3), using antiserum prepared against Tween-ether-disrupted feline leukemia virus. At the same time, reactivity to the gs-1 determinants in identical fractions was observed in complement fixation and gel diffusion assays, using guinea pig antiserum known to contain principally antibodies to rat gs-1 determinants. Presently, the principal source of rat type C viral gs antigen is rat cell line MSB, which continuously releases a rat leukemia virus pseudotype of murine sarcoma virus. The WR-9 rat virus line may be of use in providing an additional source of C-type particles that are capable of yielding good gs reagents.

Properties of oncornavirus RNA-directed DNA polymerase, the RNA template, and the intracellular products formed early during infection and cell transformation.

We have investigated three aspects of RNA turmor virus replication and cell transformation: (1) the properties of the purified avian and mammalian viral RNA-directed DNA polumerase, (2) some characteristics of the viral 60-70S RNA genome, 30-40S RNA subunits and intracellular viral RNA species, and (3) the interaction of the viral DNA polymerase with its RNA template early during infection and cell transformation by the murine sarcoma-leukemia virus (MSV[MLV]). Avian myeloblastosis virus (AMV) contains two forms of RNA-directed DNA polymerase, alpha, consisting of a single polypeptide of molecular weight 65,000, and alphabeta, consisting of two polypeptides of molecular weights 65,000 and 105,000. The alpha and alphabeta forms of AMV DNA polymerase both possess RNase H activity that requires free end termini on the ribopolymer and can degrade the RNA of the RNA-DNA hybrid in the 3' to 5' and 5' to 3' directions. But, alpha and alphabeta possess a different mode of exoribonuclease activity. While alphabeta RNase H is a processive exoribonuclease that degrades the polynucleotide chain to a core residue before attacking a second chain, alpha RNase H is a random exoribonuclease that releases the polynucleotide after each scission. Highly purified Moloney-MSV(MLV) DNA polymerase has both RNase H activity and the ability to read viral 60-70S RNA. These activities comigrate through five different steps of purification and are present at levels comparable to those found in purified AMV DNA polymerase. The MSV(MLV) 60-70S RNA genome and 35S RNA subunits were shown by periodate oxidationtritiated borohydride reduction to contain adenosine as the major 3'-terminal nucleoside. Poly (A) segments were isolated from viral 60-70S and 35S RNA by treatment with RNase A or RNase T1 and purified by afinity chromatography and gel electrophoresis. Viral poly(A) was shown to be present at the 3' terminus as -G(C,U)A190AOH. The similar sequence reported for poly(A) present in mammalian mRNA suggests that similar mechanisma are involved in the transcription and processing of both cellular and viral DNA sequences. Within transformed cells replicating MSV(MLV), viral 35S and 20S RNA were found in membrane-bound polyribosomes, whereas only 35S RNA was detected in free polyribosomes. The origin and function of 20S RNA is unknown. The early events during rapid infection and cell transformation of mouse 3T6 cells by the Harvey strain of MSV(MLV) were studied. By both autoradiographic analysis and molecular hybridization, viral DNA synthesis was detected in the cytoplasm by 1 hour after infection, reached a maximum at 2 hours, and subsequently decreased. Cytological chase experiments produced evidence that cytoplasmic viral DNA was transported to the nucleus. In situ hybridization experiments using radioactive viral DNA product as a probe demonstrated the rapid association of viral DNA sequences with the chromocenters of interphase nuclei and with the centromeric heterochromatin regions of some chromosomes.

Cerebrospinal fluid protein patterns in spasmodic torticollis.

In order to investigate the value of CSF-protein analyses in spasmodic torticollis CSF from six patients with probable organic and two patients with probable psychogenic torticollis was examined by isoelectric focusing and electrophoresis. In all the patients with organic torticollis two pathological CSF-protein fractions were found in the alkaline region on electrofocusing and in four cases aberrant fractions occurred also in the acidic pH range. An increasing number of abnormal fractions were noted during at least the first year after onset of symptoms. Lithium treatment of three patients resulted in a striking decrease of torticollis as well as of the number of abnormal CSF-protein fractions. During placebo treatment of two cases, torticollis and the pathological CSF-proteins recurred. Some observations, including a few previous autopsy findings, might indicate that an encephalitogenic agent is involved in the pathogenesis of organic torticollis. In the patients with psychogenic torticollis the CSF-protein pattern was normal. This investigation supports a recent suggestion that organic and psychogenic torticollis might be distinguished by electrofocusing of the CSF-protein.

[Studies on the production of anti-human-lymphocyte serum (ALS) in bulls].

The applicability of bulls as productive animals was considered for the preparation of anti-humans ALS. The course of immunologic response was studied by lymphoagglutination, lymphocytotoxicity, rosette inhibition, hemagglutination tests and by precipitin formation in two experimental groups immunized by different amounts of lymphocytes from peripheral blood of normal donors. The animals were found to respond well already after the second application of very small amounts of antigen (on day 0-4 times 10(7), on day 21-2 times 10(8) lymphocytes). They showed lymphoagglutination titre 1 : 512-2000, lymphocytotoxic titre being higher than 1 : 4000 and the rosette inhibition test gave a minimum titre of 1 : 65000. On the other hand, further application of a high amount of antigen (2 times 10(9), or 4 times 10(9) lymphocytes) did not lead to further increase in the titre; on the contrary - hyperimmunization resulted in a lower titre in the case of the rosette inhibition test, which is known to correlate best with the in vivo immunosuppressive activity. The hemagglutinin titre was also acceptable under the above conditions and the formation of undersirable precipitins against human serum proteins was negligible. Good response reached by a simple and economical immunization scheme speaks for the suitability of bulls for the production of ALS.

[The primary structure of a monoclonic immunoglobulin-L-chain of subgroup IV of the kappa type (Bence-Jones protein Len.)].

The complete amino acid sequence of Bence-Jones protein Len. was established by sequential analysis of tryptic and chymotryptic peptides. The result of these experiments and the comparative sequence analysis with the other 17 completely determined kappa-proteins is incompatible with the serological typisation of protein Len. as a member of subgroup II: There are 18 positions in protein Len. than cannot be associated with any one of the subgroups kappaI, kappaII or kappaIII. Also the average amino acid exchange rate between protein Len. and these subgroups is in the same range as the average amino acid exchange rate between these subgroups. Therefore Bence-Jones protein Len. is the first completely determined representative of a new IV. kappa-type subgroup. The variability of immunoglobulins follows a structural principle in which the single point mutations responsible for the variability are linked. The present paper contains the exact analysis of the linked point mutations within the so far best-investigated subgroup, kappaI (12 completely sequenced proteins). These linked exchanges allow the arrangement of the kappaI proteins in 4 subgroups and their further subdivision. The regularity of this amino acid sequence pattern can only be explained by an evolutionary origin of antibody variability. On the basis of this evolutionary mechanism the relationship of immunoglobulins can be depicted in a phylogenetic tree. Such a tree was therefore constructed for the 18 completely determined Bence-Jones proteins of kappa-type, for the first time taking into account Bence-Jones protein Len. Its topology is in complete agreement with the results of the comparative sequence analysis.

Binding of human IgE immunoglobulin to rat mast cells. A study by means of three independent techniques.

The binding of human myeloma IgE immunoglobulin on rat mast cells was studied by three independent techniques. A mixed agglutination reaction with anti-IgE-coated Sephadex granules demonstrated that only human IgE-coated rat mast cells were clearly agglutinated. This binding is strong (50% agglutination) in 3 min and progresses for 30 min (95% agglutination). Autoradiographic studies with 125I-labelled human serum proteins demonstrated the selective formation of grains on mast cells incubated with labelled IgE. Upon action of anti-IgE antiserum on IgE-coated rat mast cells, the mast cells released up to 47.5% of their total histamine content in a fluorometric histamine assay. A relationship was established between sensitizing doses of human IgE and histamine release. These results bring evidence for a binding of human IgE on rat mast cells and imply the existence of receptors for this immunoglobulin on mast cell membrane.

Gas-liquid chromatographic studies of reactions and structural relationships of steroids. Part III. 11alpha-hydroxysteroids of the androstane and pregnane series.

Qualitative and quantitative effects of classical reactions on steroids observed by gas-liquid chromatography (GLC) under standardized conditions, including the double internal-standard technique, are reported. Simple procedures applicable to nanogram amounts of reactants which afford excellent yields of the major products are described. Reactions studied include the Wolff-Kishner removal of keto groups, their conversion into hydroxyl groups with sodium-ethanol or sodium borohydride and into dioxolone derivatives with ethylene glycol; the conversion of hydroxyl into keto groups with chromium trioxide and to trimethylsilyl (TMS) ethers by hexamethyldisilazane; the hydrolysis of dioxolone and TMS derivatives by H+. Gas-liquid chromatograms of reaction mixtures of single- and multistep reactions readily provide information on the effects on the 11alpha-hydroxy and other functional groups at positions 3 and 17 (androstane series) and positions 3 and 20 (pregnane series), and the retention times of many steroids unavailable from commercial or other sources. GLC data analysis provides relationships between steroid structure and retention time from which methods for the computation of retention times and for steroid identification are designed. The accuracy of the calculation methods is demonstrated.

Serological relationship between a pathogenic strain of Marek's disease virus, its attenuated derivative and herpes virus of turkeys.

Precipitating antigens present in extracts of chick embryo cells infected with the HPRS-16 attenuated strain of Marek's disease virus (att-MDV) were separated by gel filtration on Sephadex G200 and some of their properties determined. The two main antigens detected with convalescent MD serum, referred to as 'B' and 'C' antigens, had mobilities of 0-55 and 0-25 respectively relative to phenol red on electrophoresis in 7-5% acrylamide gel. The B antigen was relatively stable and of low mol. wt. in comparison with the C antigen. B and C antigens were in some instances also detected in culture medium of infected cells, but were distinguishable from the A antigen, a major glycoprotein antigen released into the culture medium of cells infected with HPRS-16. The results of immunodiffusion studies suggest that B antigen is common to MDV and strains of herpes virus of turkeys(HVT) and that at least 2 antigens (including C) are MDV specific. The A antigen was also common to MDV and HVT strains. It was noted however that the capacity of HPRS-16/att to synthesize A antigen was considerably reduced in comparison with HPRS-16 and HVT strains, and in some preparations the A antigen could not be detected. Evidence was also obtained for the presence of HVT-specific antigens associated mainly with the cell fraction.

A revised boron-neutron capture therapy for malignant brain tumors. II. Interim clinical result with the patients excluding previous treatments.

Fifteen brain tumor patients were treated with slow neutron. It proved to extend life span of terminal glioblastoma patients irresponsive to Co-60, to 2 years, but quality of survival is poor due to complications of previous treatments. Two glioblastoma patients excluding other treatments, the only genuine Boron-neutron capture therapy cases, have been living for 39+ and 34+ months working full-scale without neurological deficit.

Thrombosis and intracranial tumors.

334 necropsy reports of intracranial neoplasm from an autopsy material over 13 years were reviewed to study the relationship of intracranial tumors to vascular thrombosis. The incidence of venous thrombosis in intracranial tumors was found to be 27.5% while that of a control group without malignancies taken at random from the autopsy material was 17%. The difference gives a statistical significance of P less than or equal to 0.05. The parameters of sex, surgical intervention, the malignancy and the histological type of the tumor apparently dod not affect thrombus formation to a statistically significant degree. There is increased thrombosis frequency with increasing age. The presence of hemiparesis or hemiparalysis does not affect the incidence of thrombosis. However, it determines to a great degree the lateralization of the thrombus.

[Measurement of attention deficit in the course of intoxications (author's transl)].

The level and course of attention was measured hourly in 9 drug intoxicated patients after a suicide attempt over periods which varied between 12 and 72 hrs. Attention was measured by the use of two additive 5 step scales for susceptibility to stimulation and reactivity, which were developed by the authors in earlier investigations and proven to be very reliable. Although, the original data set of attention measures was different among the patients, some common features could be elaborated: 1. The level of attention varies very little within 1 hr. Differences greater than one step on the scales were rarely observed between two measurements. 2. The mean course of recovery from attention deficit is linear throughout the scales while the variance is substantial at each step of the scales. For quantification of attention deficit a measure was defined which gives the relation between the actual deficit and full attention. Since the correlation between both scales is high over the whole observation period, it was concluded that the intoxication alters the level but not the structure of attention.

An electro-oculographic study of ocular bobbing and intermittent vertical oscillations occuring in the same patient.

A surviving patient with two rare types of abnormal eye movements, typical ocular bobbing and subsequently developed intermittent vertical oscillations of the eyes, is described, and a follow-up series of electro-oculographic recordings of these eye movements is analyzed. It is suggested that these two types of vertical eye movements might have a common pathophysiological background and the activity of remaining functional oculomotor pathways may play an important role in their occurrence.

[To the differential diagnosis of cranial nerve lesions: the progressive necrotising external otitis (author's transl)].

A review of necrotising external otitis, a relatively unknown and dangerous disease, brings out that, initially, it has three characteristics: a granulating necrotising ostitis of the external meatus, extreme pain and a yellowish green secretion. It is always caused by a pseudomonas infection and in almost all cases the patients suffer from diabetes mellitus. If the condition is not recognized in good time and an extensive debridement of the bone involved not performed promptly, ostomyelitis of the base of the skull may follow with involvement of cranial nerves. Severe chronic osteomyelitis of cervical vertebrae occurred in one of our cases. The neurologist must bear this disease in mind in the differential diagnosis when cranial nerves are affected because the nerve disturbances may become evident only after the local condition has subsided or the nerve deficits may be more prominent than and obscure the local ear condition. The most commonly involved nerve is the facial although there may be multiple cranial nerves involved including the third through the twelfth. If the cervical vertebrae become affected there may be nerve root lesions. A torpid meningoencephalitis may also occur. Close cooperation between otologists and neurologists is necessary to recognize and treat these conditions properly.

Detection of a TL(+) murine leukemia cell line that resists the cytotoxic effects of guinea pig complement and specific antiserum.

RADA-1 cells [H-2a thy-1b; thymus-leukemia (TL) 1, 2, 3], a radiation-induced murine leukemia cell line maintained by serial transfer in histocompatible recipients, resisted lysis by guinea pig complement (GPC) and TL 1, 3; TL 2; or TL 1, 2, 3 antiserum. The cells expressed TL antigenic specificities as determined by indirect fluorescent antibody methods, the direct isolation of TL antigens from the cells, and the capacity of the cells to reduce known titers of TL antisera. GPC was consumed to the same extent during the reaction of resistant cells and TL antisera as occurred in the reaction of sensitive cells (killed under similar conditions) and TL antisera. RADA-1 cells were not nonspecifically resistant to complement (C)-mediated lysis; they were killed in the presence of H-2a antiserum and GPC. The TL antisera contained antibodies for TL determinants. They stimulated the C-mediated lysis of ASL-1 cells (TL 1, 2, 3) and thymocytes from strain A mice (TL 1, 2, 3). The TL antigens of resistant RADA-1 cells underwent antigenic modulation, the reversible disappearance of TL antigens from the cells stimulated by specific antiserum. After the cells were treated with neuraminidase, they became susceptible to the cytotoxic effects of aliquots of the same TL antisera and GPC used previously.

Alteration of cell-surface antigenicity of the mouse plasmacytoma. I. Immunologic characterization of surface antigens masked during successive transplantations.

Alterations of membrane antigenicity of IgA-synthesizing plasmacytoma cells (58-8) induced in a BALB/c mouse were investigated with rabbit antisera against 58-8 transplanted from 7-8 generations (anti-58-8) and mouse antisera against H-2d (anti-H-2d). The 58-8 cells transplanted (TP) for 8 generations (TP8), showed moderate susceptibilities to anti-58-8 [cytotoxicity index (CI) = 72%] and to anti-H-2d (Cl = 30-40%). At TP12, the susceptibility to anti-58-8 remained (tCl approximately 50%) but there was none to anti-H-2d (Cl = 0-10%). After TP13, 58-8 cells had detectable reactivity neither with anti-58-8 nor with anti-H-2d before proteolysis. However, antigenicity was demonstrated after treatment of cells with proteases: Anti-58-8 and anti-H-2d became cytotoxic to pronase-treated 58-8 (P-58-8) and these cytotoxic reactivities of antisera could be completely absorbed with P-58-8- but not with "intact" 58-8. Anti-H-2d absorbed with P-58-8 had no cytotoxicity to BALB/c spleen cells. Absorption of anti-H-2d with BALB/c spleen cells did not show any measurable cytotoxicity to P-58-8. Appearance of antigenicity after proteolysis was transient and reversible. When P-58-8 was incubated in vitro, susceptibilities to antisera were decreased. Actinomycin D and puromycin inhibited the loss of susceptibility; these metabolic inhibitors suppressed the "masking" of antigenic determinants with protein-like material(s). In early generations (TP8), 58-8 cells were susceptible to antiserum to bone marrow-derived lymphocytes (anti-B) and complement. The cells completely absorbed the cytotoxicity of anti-B against spleen cells. After additional transplantations (TP14, TP30), susceptibilities to anti-B were no longer detectable and no removal of the cytotoxicity was shown, though the cells were pretreated with pronase. Anti-B failed to kill MOPC-31C.

Combined intra-arterial actinomycin D and radiation therapy for surgically unresectable hypernephroma.

One primary goal of preoperative radiotherapy for hypernephroma is to reduce the volume of tumor and, therefore, improve the possibility of resection. It is important that this goal be accomplished promptly so that 4 to 6 weeks after radiation therapy nephrectomy can be attempted. A longer waiting period may allow fibrosis of the normal surrounding tissues and make surgery more difficult. In addition, longer waiting periods could theoretically increase the probability of metastasis. Therefore, we plan to continue clinical investigation on the use of combined intra-arterial actinomycin D and radiotherapy as a possible useful means of improving the possibility of prompt surgical resection, since theoretically this regimen may be a method of increasing the effective radiation dose to the hypernephroma without increasing the effective radiation dose to surrounding normal tissue, such as bowel. The method may also have merit as an improved means of palliating selected patients with metastases who are symptomatic from a bulky primary hypernephroma.

Human glomerular cells in tissue culture.

Cells from human glomeruli explanted in tissue culture were grown and subcultivated up to 12 to 13 times. Light and electron microscopic studies revealed these cells to be morphologically distinct from fibroblasts. By electron microscopy, an extracellular material resembling basal lamina was seen and prominent intracellular microfilaments were evident. Immunofluorescent microscopy demonstrated reactivity of heterologous antiglomerular basement membrane antibody with aggregates of extracellular material. Absorption experiments using antiglomerular basement membrane antibody showed that the extracellular materiial shared some antigenic components with glomerular basement membrane. Antibody to cultured glomerular cells stained the mesangium and glomerular basement membrane of normal human kidney. This antibody was nephrotoxic in monkeys, induced proteinuria with proliferative glomerulonephritis, and localized to the mesangium and glomerular basement membrane of monkey glomeruli. These findings and the presence of prominent intracellular microfilaments (contractile elements) suggest that the glomerular cells may be of mesangial origin.

Casual blood-ethanol estimations in patients with chronic liver disease.

Patients attending a clinic for diseases of the liver were tested for blood-ethanol by a gas chromatographic technique sensitive to about 5 mg/dl (1 mmol/1). Of 172 patients (51 men, 121 women) 36% gave a history of heavy drinking (greater than 80 g ethanol/day; equivalent to 8 fl oz of whisky or 1 litre of wine) and 13% had ethanol in the bloodstream at values of 8-400 mg/dl. 42 patients (24%) had the liver-biopsy changes of alcoholic liver disease, and 17 of these had ethanol in the blood at one time or another. Nearly half (22/49) of all patients admitting heavy drinking also had detectable blood-ethanol. In all cases but 1 where blood-ethanol was found, a drinking history was admitted on first attendance, and alcoholic liver disease was nearly always found on subsequent biopsy. Blood-ethanol and admission of drinking were most constantly found in association with alcoholic steatosis and hepatitis. Both features were less commonly present in cases of alcoholic cirrhosis. Only 1 patient of 22 with "cryptogenic" cirrhosis on biopsy was found to have both ethanol in the blood and an alcoholic history, although 5 had an alcoholic history alone. The value of serial blood-ethanol estimations in the treatment of alcoholics and the detection of relapses is demonstrated. The findings confirm the relatively low frequency of alcoholism as a contributor to cirrhosis in the United Kingdom. Alcohol does not seem a major cause of cryptogenic cirrhosis. Casual blood-ethanol estimation is a useful and objective adjunct to techniques of investigating diseases of the liver.

Secretory component and sudden-infant-death syndrome.

Post-mortem specimens of blood, respiratory-tract washings, bronchopulmonary tissue, spleen, and thymus were examined for respiratory viruses, immunoglobulins, and secretory component (S.C.) in eight infants with sudden-infant-death syndrome (S.I.D.S) and in eight other (control) infants with an identifiable cause of death. Serum-immunoglobulin levels were similar in infants with S.I.D.S. and in control infants. In some S.I.D.S. cases serum-IgM was slightly raised. Respiratory syncytial virus was found in the pulmonary tissues of five S.I.D.S. patients but no viruses were isolated from other subjects. However, in one control subject parainfluenza type-3 viral antigen was detected in the bronchial tissue. In all patients with S.I.D.S., immunological reactions and fluorescent antibody staining for S.C. in the broncho-pulmonary epithelium were absent or grossly reduced. The levels of IgG and IgM in bronchial washings were unremarkable. These observations suggest a possible defect in respiratory mucosal defence in patients with S.I.D.S.

Bilateral Wilms' tumour. Age at diagnosis, associated congenital anormalies, and possible pattern of inheritance.

A series of 87 patients with Wilms' tumour seen during the period 1960-73 included 11 (13%) with bilateral tumours. 6 patients presented with simultaneous bilateral tumours, 2 had tumours in each side of a horseshoe kidney, and 3 later developed a tumour in the remaining kidney. There was no reported familial incidence of Wilms' tumour. Maternal age at birth of the patients with simultaneous bilateral tumours was over 30 years in 7/8 cases. The average of patients with bilateral tumours was 15 months, whereas that of patients with unilateral tumours was 31/2 years. All the simultaneously occurring bilateral tumours and those within a horseshoe kidney were multifocal, whilst the sequentially occurring bilateral tumours and the unilateral tumours all developed as a single tumour mass within the affected kidney. Associated congenital anomalies were found in 5 (45%) of 11 patients with bilateral tumours, several of whom had more than one defect. Of 76 patients with a unilateral tumour, only 3 (4%) had congenital anomalies.

Liver damage after paracetamol overdose. Comparison of liver-function tests, fasting serum bile acids, and liver histology.

54 patients have been studied after paracetamol (acetaminophen) overdose. Liver-function tests and fasting serum bile-acids were measured daily; liver biopsy was done in all cases, and slides were examined "blind" to assess liver damage. The plasma-paracetamol was measured on one occasion. A histological abnormality was present in the livers of 53 of the 54 patients, and was minor in 23, moderate in 16, and severe in 14. In 6 patients with moderate and 18 with mild histological abnormality liver-function tests were normal. A serum-aspartate-aminotransferase above 400 units/1 was always associated with severe histological liver damage. Fasting serum bile-acids were raised in 51 of the patients with abnormal liver histology; the serum-bile-acid seemed to be a more sensitive indicator of mild liver-cell damage than was the transaminase level. There was, however, little correlation between increase in bile-acid concentration and the degree of histological abnormality. As a result of these investigations empirically determined levels of plasma-paracetamol have been drawn which give a guide to the likelihood of liver damage after paracetamol overdose.

The utilization of adenosine triphosphate in rat mast cells during histamine release induced by anaphylactic reaction and compound 48/80.

The ATP content of rat peritoneal mast cells has been studied in relation to histamine release induced by compound 48/80 and antigen-antibody (anaphylactic) reaction in vitro. When the ATP content of actively sensitized mast cells was reduced to different levels by oligomycin, a good correlation was obtained between the ATP levels and the amounts of histamine released by the anaphylactic reaction. A similar linear relation has previously been demonstrated between the ATP levels of mast cells and histamine release induced by compound 48/80. The ATP content of mast cells was also studied at different intervals after the exposure of the cells to antigen or compound 48/80. No significant change in the ATP content was observed in untreated mast cells during the short period when histamine release occurs. If, however, the mast cells were preincubated with oligomycin or 2-deoxyglucose to reduce the rate of ATP synthesis while a large part of the histamine release remained unaffected-a decrease in the ATP content could be demonstrated in close time relation to both anaphylactic and compound 48/80-induced histamine release. The observations indicate an increased utilization of ATP in mast cells during the release process.

[A new method of transseptal levocardiography].

Perforations of the left atrial or ventricular wall and extravasations of contrast medium during transseptal left heart catheterization or angiocardiography can be eliminated by replacing the routinely used transseptal catheters with Pigtail-catheters. With 2.8% minor complications without sequelas in 181 successful studies, transseptal angiocardiography of the left heart through Pigtail-catheters is not only less hazardous than injections through the transseptal catheters employed up to now, but bears even less risk than direct retrograde injection into the left ventricle. To show the left atrial cavity and the mitral valve, transseptal left atrial injection is the method of choice. For quantitative angiocardiography to evaluate left ventricular function, transseptal angiocardiography with injection into the left atrium is superior to retrograde direct ventriculography in our experience, as ventricular ectopic beats were absent and supraventricular ectopic beats as rare as 5% of the cases, local disturbances of wall motion during injection could be avoided and 1 or 2 more cycles could be evaluated before the depressant effect of contrast medium started.

Feulgen-DNA-cytophotometry on mammary tumor cells from aspiration biopsy smears.

The nuclear DNA content of cells from 20 cases of mammary aspirates was estimated by microspectrophotometry. Smears stained according to Papanicolaou were destained and Feulgen-reacted. Among the smears there were 14 with doubtless tumor cells, four with a benign cell picture, and two of a doubtful nature. The mammary lesions were in all cases verfied by histology. Measurements showed that smears of benign tumors had a diploid or diploid to tetraploid distribution of DNA. The two doubtful smears also showed the same DNA pattern, although in one case the cells were from a papillary carcinoma and in the second case from a fibroadenoma. Malignomas with a cell picture, monomorphous-atypical in Papanicolaou staining exhibited a DNA histogram with a peak in the diploid to hyperdiploid ranges. Carcinomas showing a polymorphous-atypical cell picture were characterized either by polyploid values, by a large spread, or by aneuploidy. Hence, it is impossible to safely distinguish benign cases from carcinomas by the DNA pattern, since DNA distribution is fairly identical in part of the cases in either group.