Differentiation of malignant myoblasts in 7,12-dimethyl-benz(a)-anthracene-induced rhabdomyoblastomas. An electron microscopic study.

The differentiation of DMBA-induced rhabdomyoblastomas was studied during the early stages of tumour growth. Two cell populations were found to constitute the tumour tissue: small cells (SC) and long spindle-shaped cells (LSSC). The SC were the only tumor cells at the earliest detectable stage of tumour growth 10 weeks after intramuscular injection of DMBA. They had small heterochromatic nuclei with a compact nucleolus containing only fibrillar components. The cytoplasm was very rich in SER of tubular type, dense bodies and Golgi apparatus. Centrioles at all stages of the replicative cycle were very frequently observed. The cells did not fuse and showed no tendency to differentiate. The LSSC had large euchromatic nuclei with multiple irregular nucleoli containing both fibrillar and granular components. The cytoplasm had an abundant GER and well-developed Golgi apparatus. These cells formed 100 Angstrom thick cytofilaments the increase of which paralleled reduction of GER. The cells tended to fuse but did not form myofibrils. A rare variant of these cells neither possessed Golgi apparatus nor formed cytofilaments but accumulated dense protein substance in the cisternae of the GER. Myotubes with cross-striated myofibrils were but occasionally observed. The ultrastructural characteristics of both cell types revealed essential differences in the biosynthetic activity and the degree of differentiation. The SC were considered to belong to the myogenic cell line and to be most probably the malignant counterpart of proliferating satellite cells (presumptive myoblasts) and precursors of the LSSC. Morphologically and developmentally the LSSC bore close resemblance to normal myoblasts but the proliferative capacity of some of them seemed to be lost. The differentiation of the malignant myoblasts in the DMBA-induced rhabdomyoblastomas was similar to the early differentiation of the normal muscle tissue.


Insulin and glucagon responses of transplanted intrasplenic pancreatic islets.

Two transplant procedures have been investigated in which one third of the pancreas was autotransplanted into the splenic pulp of dogs. The two procedures consist of simple mechanical dissociation of the pancreas or mechanical dissociation followed by collagenase digestion. The ability of the endocrine segment of the transplant to survive and function was assessed by stimulation with arginine and measurement of insulin and glucagon response. The results demonstrate that both transplant procedures result in functioning beta and alpha cells that rapidly secrete both insulin and glucagon in response to arginine stimulation. However, greater insulin responses were obtained when mechanically dissociated but nonenzyme digested pancreatic tissue was used for transplantation. The spleen appears to be an excellent transplant site for the reception of endocrine pancreatic tissue and allowed both beta and alpha cells to survive following transplantation.


[Phylo- and ontogenetic development of the nucleus of the brachium of the inferior colliculus].

In recent neuromorphological investigations and in studying the passage, as an addition to nuclei of the auditory system so called "parabrachial area" was subjected to a special comparative cytoarchitectonic investigation and terminological definition, since the term "parabrachial nucleus" has already been reserved in neuromorphology to designate other brain formations. The investigation has demonstrated that the term "nucleus of the inferior quadrigemina brachium" is more suitable to the form studied. This nucleus has been stated to penetrate deeply into the inferior tuberculum corpori quadrigemina, unlike the previous notion according to which it was believed to be situated between the internal geniculate body and the inferior tuberculum corpori quadrigemina. Therefore, two parts have been distinguished in the formation in question, inherent to all the mammalia studied, including the man. One of them--extracollicular portion, though it has not been previously studied from the view of comparative architectonics, nevertheless, it has been described in literature. The other--intracollicular portion is described for the first time in the work. In spite of a close topographic-anatomical proximity with the inferior corpora quadrigemina, the intracollicular portion of the nucleus cannot be a derivative from the latter.


Posterior corneal crystalline deposits in benign monoclonal gammopathy: a clinicopathologic case report.

A 74-year-old woman had bilateral, deep stromal, patchy crystalline corneal deposits with the greatest density in the midperiphery. Visual acuity was 6/120 in the right eye and finger counting at 1 m in the left eye. Histological examination of the corneal button showed large, irregular amorphous masses in the posterior stroma. The deposits stained red with Masson's trichrome and were positive for protein with the Danielli stain. Stains for amyloid, copper, and lipid were negative. The immunoperoxidase stain was positive for polyvalent IgG and kappa light chains. Transmission electron microscopy disclosed electron-dense deposits with linear and honeycomb profiles. Laboratory investigations disclosed elevated serum and urinary IgG kappa light chain (Bence Jones protein) levels. Urinary amino acids were normal. The serum copper level was elevated. Antinuclear antibody was positive at a titer of 1:80. A bone marrow aspirate was normal, as were roentgenograms of the skull.


Mean circulation time of fluorescein in retinal vascular segments.

Mean circulation time (MCT) of sodium fluorescein in retinal vascular segments was determined in healthy human volunteers, using the dye dilution technique. The fluorescein dilution curves were recorded with a two-point fluorophotometer. Average MCT was 3.9 seconds. The accuracy of the method was determined from repeated measurements in owl monkeys and human volunteers. No significant linear correlation was found between the MCT and the cross section of the vessels feeding and those draining the segment. The application of this technique to clinical work is discussed, and is illustrated by three clinical cases.


Isolation, characterization and pathology of the toxin from a Microcystis aeruginosa (= Anacystis cyanea) bloom.

The nature of the toxicity of a bloom of blue-green alga, M. aeruginosa (= Anacystis cyanea), that occurred in a man-made lake was investigated. Crude algal bloom extracts were toxic to laboratory mice when injected intraperitoneally. The lethal dose (LD100) of these extracts was 15-30 mg of lyophilized algal bloom per kilogram body weight. The toxin was purified by a procedure that included ammonium sulphate fractionation, solvent extraction, acid precipitation, Sephadex G25 and DEAE-Sephadex chromatography, and high-voltage electrophoresis at pH 6.5. The preparation gave a single spot on high-voltage electrophoresis at pH 9.0, had no free amino group, and was characterized by a simple amino acid composition of equimolar quantities of L-methionine, L-tyrosine, D-alanine, D-glutamic acid, erythro beta-methyl aspartic acid and methylamine. The LD50 for the purified toxin was estimated to be 0.056 mg/kg of mice, and the approximate LD100 is 0.070 mg/kg, based on the total material found from amino acid analysis. Parenteral administration of the purified toxin to mice produced extensive liver lobular haemorrhage and death within 1-3 h. Repeated inoculation of sublethal doses daily over some weeks produced progressive hepatocyte degeneration and necrosis and the development of fine hepatic fibrosis.


Distribution of surface IgM and IgD on single cells from lymphocyte subpopulations.

The surface immunoglobulin heavy chains on individual spleen cells fractionated by velocity sedimentation were studied using fluorescent antisera. In adult mice, cells bearing both mu and delta chains were found in all fractions. While there was an increase in the proportion of cells bearing mu only in the medium to large cell fractions, the majority of cells bearing mu only were small lymphocytes. Results obtained using 3-week-old mice were basically similar, but showed both a marked decrease in small mu + delta + cells and a marked increase in small mu + delta - cells when compared with adult animals.


Influence of beta-alanine on mating and territorialism in Drosophila melanogaster.

Effects of beta-alanine on mating behavior and aggression were studied in Drosophila melanogaster using the following competitive pairs: (1) homozygous black (b/b) flies, in which beta-alanine synthesis is decreased, vs. alanine is blocked vs. wild-type (e+/e+) flies; (2) dark flies, in which beta-alanine incorporation is reduced, owing mainly to chromosome 3, vs. light flies collected from the same population as were the dark flies; (3) homozygous black (b/b) flies, in which beta-alanine synthesis is decreased, vs. beta-alanine-infected b/b flies, which are phenocopies of wild-type flies. The behavior of mixed-sex groups was studied in a large, illumination-graded observation chamber containing food and in small uniformly illuminated cells also containing food. The relative competitive mating abilities of these types were measured in both experimental conditions. Uninjected black flies, but not injected ones, showed weak and unsteady gait and weak wing extension. In ebony these abnormalities were more extreme. Dark flies did not show these abnormalities. Accelerated sexual maturation was indicated in males by early onset of courtship and enhanced territorial aggression and in females by earliness of mating. Such acceleration was observed in ebony and dark flies, compared with light flies, and among beta-alanine-injected b/b flies competing with uninjected black flies. Ebony males, although maturing earlier than wild-type males, were less successful than wild-type males in mating. This difference was even greater when the flies were all allowed to mature before competing. Ebony females outmated wild-type females. Dark flies outmated light flies, and beta-alanine-injected b/b males outmated uninjected black males, especially in bright light. Ebony flies mated much longer than wild-type flies, and black flies mated slightly longer than injected b/b flies. There was some spatial isolation of ebony from wild type, dark from light, and beta-alanine-injected from uninjected b/b flies in the illumination-graded observation chamber. Ebony flies more than wild type concentrated near food. Flies were attracted to the current of moist inlet air. They were also attracted to deposited excrement, and males defended such deposits as a mating area, thus showing rudiments of arena behavior in which a mating area away from the oviposition site is defended. Usually, however, the defended area focused on food.


Hemodynamic and myocardial effects of long-lasting venodilation in the conscious dog: analysis of molsidomine in comparison with nitrates.

The effect of molsidomine-induced venodilation on cardiac preload was studied in conscious resting dogs, instrumented to analyze left ventricular function and myocardial perfusion. Direct effects on veins were studied during chloralose anesthesia by measuring regional venous capacitance changes with an induction angiometer. Kinetics of molsidomine-induced effects were compared to those induced by nitroglycerin and isosorbide dinitrate. This comparison was restricted to low i.v. dosages, causing only transient threshold effects on peripheral resistance and heart rate. During molsidomine-induced venous pooling, neither any direct effect on the coronary circulation nor any direct cardiac depressant activity of the drug was detected. 100 microgram/kg molsidomine caused a reduction of left ventricular preload by 5 mm Hg, lasting at least 4 hours. This effect was significantly more pronounced than that induced by 1 microgram/kg nitroglycerin or by 25 microgram/kg isosorbide dinitrate, lasting 2 min or 20 min, respectively. However, in raising regional venous capacitance, these nitrate dosages were equi-effective to 100 microgram/kg molsidomine, the effect of which was persistent and with a greater delay in onset. These results indicate that the lasting persistance of venodilation is a decisive factor for the amount of volume pooled in the capacitance system and, consequently, for the extent of preload reduction obtained. It is concluded, that lasting vasodilation, restricted to the veins, is beneficial for ventricular performance in ischemic heart disease.


Genetics of esterases in Drosophila. VI. Gene system regulating the phenotypic expression of the organ-specific esterase in Drosophila virilis.

It is shown that the gene controlling the synthesis of the organ-specific S-esterase of Drosophila virilis ejaculatory bulbs is located on the second chromosome (at approximate position 192.1 +/- map units). The cells of the genital imaginal disks are determined for the synthesis of S-esterase 10-12 hr after the second molt. The organ-specific esterase can be detected after adult emergence only. It is preceded by an increase in RNA content and by enhancement of RNA synthesis in the cells of the ejaculatory bulbs. Interstock differences were found in the level of the activity of S-esterase, which is under the control of the X chromosome, as well as in the time of expression of enzyme activity, which is controlled by the fifth chromosome. It is suggested that the specific phenotypic expression of this enzyme depends on the system of genes with regulatory expression at both the transcriptional and posttranscriptional levels. The genetic control of the synthesis of the S-esterase described is a convenient model for studying mechanisms of gene activity regulation in eukaryotes.


Effects of nitroglycerin and dipyridamole on regional myocardial blood flow.

The effects of intravenous administration of nitroglycerin or dipyridamole on distribution of myocardial blood flow in the heart after coronary underperfusion were investigated. In 18 open chest, anesthetized dogs, flow of the tubing supplying the left circumflex coronary artery (LCA) was reduced and kept constant, and then the drug was given intravenously. Before and after an administration of saline, nitroglycerin (0.04 mg/kg) and dipyridamole (0.2 mg/kg), myocardial blood flow was measured by tracer microspheres (TM). The first and second carbonized 15 mu spheres (TM1 and TM2) were injected into the left atrium and the cannulated LCA perfusion system respectively before medication. Similarly, the third and fourth spheres were given after drug. Thus, by using four different TMs (141Ce, 51Cr, 85Sr, 46Sc) regional myocardial blood flow from the LCA perfusion route (TM2 and TM4) was measured separately from that of the other normally perfused coronary route (TM1 and TM3). The effect of drugs on flow distribution of these two independent routes was measured. The LCA underperfusion produced a significant diminution of regional flow and its end/epi ratio in the LCA territory. Nitroglycerin increased end/epi ratio in the ischemic myocardium in the LCA perfusion route, but dipyridamole increased the normally perfused coronary flow but caused a shift of flow from the ischemic left ventricle to the atria and right ventricle within the LCA territory.


The effect of nitroglycerin on myocardial release of inosine, hypoxanthine and lactate during pacing-induced angina.

The efficacy of nitroglycerin as an antianginal drug has been evaluated by calculation of myocardial extraction and production values of lactate and the adenosine triphosphate (ATP) catabolites inosine and hypoxanthine. Coronary venous and arterial blood was sampled at rest, during pacing-induced angina and 4--6 min after nitroglycerin at identical paced heart rates for enzymatic assay of inosine and hypoxanthine after separation by column chromatography and for determination of lactate. Sublingual nitroglycerin given to 10 patients with coronary artery disease decreased coronary venous lactate values from 1175 +/- 320 mumol/l during pacing-induced angina to 950 +/- 240 mumol/l (p less than 0.05). The calculated myocardial lactate production during angina (-31 +/- 19%) diminished after nitroglycerin (-1.7 +/- 22%) (p less than 0.0025). Coronary venous inosine values during angina (1275 +/- 865 nmol/l) decreased after nitroglycerin (795 +/- 555 nmol/l) (p = n.s.), the arterial values (885 +/- 610 nmol/l) increased (960 +/- 580 nmol/l) (p = n.s.), the myocardial inosine release (-26 +/- 20%) changed to extraction values (19 +/- 19%) (p less than 0.0005). Coronary venous hypoxanthine values during angina (1540 +/- 1035 nmol/l) were reduced (1110 +/- 675 nmol/l) (p = n.s.); the arterial values (1625 +/- 1050 nmol/l) decreased (1510 +/- 935 nmol/l) (p = n.s.), the myocardial hypoxanthine extraction (0.3 +/- 29%) with a wide individual variability increased after nitroglycerin (24 +/- 13%) (p less than 0.025). The myocardial release of inosine and lactate during severe angina with significant positive correlation (r = 0.66, p less than 0.0025) demonstrates that anaerobic glycolysis is accompanied by ATP breakdown. The unchanged myocardial inosine and hypoxanthine extraction after nitroglycerin indicates that nitroglycerin is capable of attenuating this effect. In spite of reduced mean myocardial lactate production after nitroglycerin ischemic myocardial energy deficiency may be less marked. Thus, the enhanced myocardial inosine uptake may be one factor contributing the beneficial effects of nitroglycerin including the improvement of myocardial oxygen balance.


Alcohol-oxidizing enzymes in 13 Drosophila species.

Starch and polyacrylamide gel electrophoresis were used to ascertain the substrate specificities of alcohol-oxidizing enzymes in 13 Drosophila species. The substrates used were a variety of long- and short-chain aliphatic alcohols, one aromatic alcohol, and benzaldehyde. Only one enzyme (product of a single-gene locus) showed significant NAD+-dependent alcohol dehydrogenase activity with short-chain aliphatic alcohols. The 13 species, belonging to four different Drosophila groups, all showed a similar complement of alcohol-oxidizing enzymes, although differences in electrophoretic mobility and in levels of activity existed from species to species. These findings are relevant to the adaptation of Drosophila to alcohol environments.


Heat stability studies at the alpha-glycerophosphate dehydrogenase locus in populations of Drosophila melanogaster.

The level of hidden variation in populations of Drosophila melanogaster at the Gpdh+ locus was determined by thermal stability studies of the protein. The results indicate a lack of variation using these methods both in and between the two common electrophoretic variants. It is suggested that alpha-GPDH is conserved in primary structure, which may be related to its critical role in flight muscle metabolism.


Calcium accumulation during sporulation of Bacillus megaterium KM.

Accumulation of Ca2+ in Bacilli occurs during stages IV to VI of sporulation. Ca2+ uptake into the sporangium was investigated in Bacillus megaterium KM in protoplasts prepared in stage III of sporulation and cultured to continue sporulation. These protoplasts and whole cells exhibit essentially identical Ca2+ uptake, which is compared with that of forespores isolated in stage V of sporulation. Ca2+, uptake into both sporangial protoplasts and isolated forespores occurs by Ca2+-specific carrier-mediated processes. However, protoplasts exhibit a Km value of 31 micrometer, and forespores have a Km value of 2.1 mM. Sporangial protoplasts accumulate Ca2+ against a concentration gradient. In contrast, Ca2+ uptake into isolated forespores is consistent with downhill transfer in which both rate and extent of uptake are affected by the external Ca2+ concontration. Dipicolinic acid has no effect on Ca2+ uptake by isolated forespores, apart from decreasing the external Ca2+ concentration by chelation. A model for sporulation-specific Ca2+ accumulation is proposed, in which Ca2+ is transported into the sporangium, resulting in a concentration of 3--9 mM in the mother-cell cytoplasm. This high concentration of Ca2+ enables carrier-mediated transfer down a concentration gradient into the forespore compartment, where a low free Ca2+ concentration is maintained by complexing with dipicolinic acid.


Functional involvement of carbonic anhydrase in calcium transport of the chick chorioallantoic membrane.

Carbonic anhydrase activity was demonstrated in the chick-embryonic chorioallantoic membrane and was correlated with the Ca2+-transport activity of the membrane. It is inhibited by sulphonamides and is expressed in the chorioallantoic membrane in an age-dependent fashion during embryonic development. Ca2+ uptake by the chorioallantoic membrane in vivo also increases in a similar age-dependent manner. The temporal increase in these activities is coincident with calcium deposition in the embryonic skeleton. Incubation of the chorioallantoic membrane in ovo with sulphonamides specifically inhibits both the carbonic anhydrase and the Ca2+ uptake activities of the membrane in vivo. Enzyme histochemistry revealed the carbonic anhydrase activity is localized in the Ca2+-transporting ectodermal cells of the chorioallantoic membrane. These results, taken together, indicate that carbonic anhydrase may be functionally important in the Ca2+-transport activity of the chorioallantoic membrane.


Serum amyloid A protein in amyloidosis, rheumatic, and enoplastic diseases.

Serum levels of amyloid protein A (SAA) have been shown to be elevated in different types of amyloidosis and in rheumatic diseases by radioimmunoassay using 125 iodine labeled AA and anti-AA. SAA levels were elevated in both primary and secondary amyloidosis, but there were highly significant differences between these levels. In heredofamilial amyloid, SAA levels were within normal limits. While the mean SAA level was elevated in persons over 70 years, the fact that some persons in this age group had normal levels suggested that marked elevation after age 70 may be due to occult inflammatory or neoplastic disease. High SAA levels in patients with rheumatoid arthritis correlated, in most cases, with physician evaluation of disease activity and Westergren ESR. SAA levels in patients with systemic lupus erythematosus were lower than those in patients with rheumatoid arthritis, and most patients with degenerative joint disease had normal levels. Very high levels of SAA were found in patients with neoplastic diseases. Patients with carcinoma of the lung and bowel had much higher levels than patients with carcinoma of the breast. Determination of SAA levels may be of value in evaluating different forms of systemic amyloidosis, assessing the activity of rheumatic disease, and screening for occult inflammatory or neoplastic disease.


Significance of serum C-reactive protein elevation in patients with systemic lupus erythematosus.

Serum C-reactive protein (CRP) concentration was determined by a radial immunodiffusion method in serum samples collected over a mean period of 19 months from a group of 17 patients with systemic lupus erythematosus (SLE). Thirty-two episodes of CRP elevation were detected. Twenty of these were associated with active SLE, and 9 with proven or possible infection or bone fracture. In 3 cases no explanation for CRP elevation was readily apparent. There were 8 instances of onset or exacerbation of lupus activity without accompanying CRP elevation. These data indicate that CRP elevation in the course of SLE is frequently associated with activation of lupus, and that detection of such elevation does not differentiate between lupus activity and infection.


Antibodies reacting with ribosomal ribonucleoprotein in connective tissue diseases.

Precipitating antibody to an antigen present in cytoplasm was detected in the sera of 7 patients with systemic lupus erythematosus, 1 patient with an overlap toid arthritis, and 1 patient with mixed connective tissue disease. By physicochemical and enzymatic studies, the antigen was shown to have the properties of ribosomal ribonucleoprotein (gamma-RNP). Cytoplasmic staining in immunofluorescence was observed with all the 9 sera containing antibodies to gamma-RNP. In certain cases, cytoplasmic staining was associated with nucleolar staining. Antibody to gamma-RNP is different in immunologic specificity and clinical significance from antibody to nuclear RNP and is present primarily in patients with systemic lupus erythematosus.


Treatment of established atherosclerosis during cholesterol feeding in monkeys.

A semipurified diet containing 43% of the calories as fat and 1.2 mg of cholesterol/cal was fed to cynomolgus monkeys (Macaca fascicularis) for 6 months; the cholesterol content was reduced to 0.34 mg/cal for the next 18 months. During the latter period, the monkeys were assigned to 4 groups of 18 animals each and received the following dietary additions: A, none (controls); B, cholestyramine (5%, w/w); C, dextrothyroxine (0.003%); and D, Wy-14,643 (0.45%). Cholestyramine normalized plasma lipid levels and reduced the size of aortic and coronary atherosclerotic lesions in spite of the high-fat, high-cholesterol intake. Dextrothyroxine reduced cholesterolemia but did not modify the extent of arterial lesions. Wy-14,643 changed neither plasma cholesterol levels nor the extent of atherosclerosis.


The relation of hypercholesterolemic fatty streaks to intimal permeability changes shown by Evans blue.

The aortic localization of diet-induced fatty streaks in relation to focal increases in intimal permeability was evaluated in cynomoglus monkeys. Animals fed a hypercholesterolemic diet and studied at 10, 15, and 100 days had increasing intensity of Evans blue dye uptake. The overlap of fatty streaks with areas of dye intake increased as the areas of dye uptake enlarged, but all hypercholesterolemic groups showed some fatty streaks not topographically related to areas of dye uptake or flow instability. Because the upper thoracic aorta tended to show more advanced fatty streak formation dissociated from evident permeability change or hydraulic instability, it is suggested that mechanical factors associated with the geometric configuration of the descending thoracic aorta may have a significant role in the localization of some hypercholesterolemic fatty streaks.


[Immunodeficiencies and enzyme deficiencies].

The first part of the paper outlines the actual situation of the recent investigations on the interpretative problem of the immunological deficits combined with enzymatic deficits. ADA deficit associated with combined immunodeficiency is an autosomal recessive form. Low enzyme levels, which is produced by a structural gene located on chromosome 20, were detected in chronic lymphatic leukemias where ADA decrease is correlated to low levels of T lymphocytes and the prevalence of an atypical B lymphocyte population. Particularly high levels of ADA were detected in acute lymphoblastic leukemias, in transplant rejection, in blastic crisis of chronic myeloid leukemia... NP deficit is associated with a T branch immunodeficiency, with high levels of inosine, guanosine, hypouricemia, hypochromic microcythemia and hematopoietic tissue megaloblastosis. This enzyme, with trimeric structure, whose structural gene is located on chromosome 14, shows a cytoplasmic location, and its maximum activity is to be found in T lymphocytes separated by rosetting. IGPT deficit is to be held responsible for the neurological Lesch-Nyhan syndrome. This deficit is associated with a depression of B lymphocyte function evaluated as response to the mitogens (PWM, Protein A) or as specific immunoglobulins production. At last the Authors report some personal investigations performed on hemolysates and lymphocytes of subjects with impaired immunity, as well as on some children at birth to establish the correlation between ADA and NP behaviour and immunosurveillance. Lastly, the data on the variations of the enzymatic and immunological parameters of subjects with immunodeficiency associated to enzymopenia after red cell transfusion are reported.


Immunological heterogeneity of haemophilia B: a multicentre study of 98 kindreds.

An electroimmunoassay with a precipitating rabbit anti-human factor IX antiserum and an inhibitor neutralization assay with a non-precipitating homologous antibody were used to measure factor IX antigen (IX:Ag) in 117 patients from 98 kindreds with haemophilia B; and to investigate in a mixed population the incidence of different immunological types of the disease. Although the two assays showed an excellent correlation, the electroimmunoassay was selected for its simplicity as a criterion for classification. 52 kindreds, referred to as haemophilia B-, were characterized by severe deficiency of factor IX coagulant activity (less than 0.01--0.03 u/ml) and unmeasurable IX:Ag (less than 0.12 u/ml): this genetic variant of the disease appears to be related to a complete or marked suppression of factor IX synthesis. In 16 kindreds, a severe or moderately severe IX:C deficiency was associated with normal or increased levels of IX:Ag (haemophilia B+): among them, a subgroup of five kindreds could be identified by the additional abnormality of a prolonged Thrombotest clotting time (haemophilia BM). These patients are likely to be the expression of normal or increased synthesis of a factor IX molecule markedly defective in the site(s) responsible for coagulant activity. Reduced levels of IX:Ag (0.12--0.65 u/ml, characterized the remaining 30 kindreds, presenting with IX:C levels ranging from less than 0.01 to 0.21 mu/ml. In 28 there was a significant excess of IX:Ag over IX:C, suggesting a reduced capacity to synthesize the factor IX molecule accompanied by a variable defect in the coagulant site; the remaining two kindreds, which showed a concomitant reduction of IX:C and IX:Ag, are likely to be examples of a true reduction of factor IX synthesis.


Enzymatic and chemical properties of an endopeptidase from the larva of the hornet Vespa crabro.

An endopeptidase from the larvae of the hornet Vespa crabro has been purified to homogeneity. The enzyme has been characterized with respect to molecular weight, amino acid compositon, and amino- and carboxyl-terminal sequences. The catalytic properties of the hornet protease are similar to those of bovine chymotrypsin with respect to inactivation by phenylmethanesulfonyl fluoride and carbobenzoxyphenylalanine chloro ketone and preferential peptide bond cleavage at aromatic amino acid residues. In contrast to bovine chymotrypsin, the hornet protease is not inhibited by the basic pancreatic Kunitz inhibitor, soybean inhibitor, or chicken ovomucoid. The molecular weight, as determined by several independent methods, was found to be 14 500. The protease is a single-chain protein containing two disulfide bonds. The terminal sequences are: NH2-Ile-Val-Gly-Gly-Ile-Asp.....Gly-Lys-Tyr-Pro-Tyr-Gln-Val-Ser-Leu-Arg-COOH.


Isolation of a high specific activity 35S ribosomal RNA precursor from Tetrahymena pyriformis and identification of its 5' terminus, pppAp.

RNA isolated from starved and refed Tetrahymena pyriformis GL, pulse labeled with H3(32)PO4, contains an RNA molecule of 2.4 x 10(6) molecular weight which can be identified by polyacrylamide gel electrophoresis. This RNA has been shown to be a precursor of the mature 25S rRNA and 17S rRNA by a kinetic analysis of its synthesis, by hybridization competition with pure 25S rRNA and 17s rRNA, and by two-dimensional fingerprint analysis. The 5'-terminal nucleotide was isolated from the total digestion products of the 35S rRNA precusor and shown to be pppAp by analysis of the products of snake venom phosphodiesterase digestion. Low, but significant yields of the 5' terminus pppAp were also isolated from the 17S rRNA demonstrating that the primary transcript of the rRNA gene in T. pyriformis contains the 17S rRNA at its 5' end.


Probes of beta-galactosidase structure with antibodies. Reaction of anti-peptide antibodies against native enzyme.

Antibodies were prepared against 18 tryptic and cyanogen bromide peptides from beta-galactosidase ranging in size from 15 to 96 amino acid residues representing more than 80% of the polypeptide chain. They were tested for binding capacity and affinity toward their homologous antigens and toward the whole native protein. Nine antisera bound to beta-galactosidase; these had been raised against certain peptides from the central and carboxyl-terminal regions of the poly-peptide chain. Based on these results a preliminary model of the three-dimensional structure of the folded protein is suggested.


Probe of beta-galactosidase structure with iodoacetate. Differential reactivity of thiol groups in wild-type and mutant forms of beta-galactosidase.

Carboxymethylation with 14 C-labeled iodoacetate of cysteine residues in wild-type beta-galactosidase from Escherichia coli and in a defective beta-galactosidase from deletion mutant strain M15 was investigated in order to determine accessible positions in the tetrameric wild-type form and the dimeric mutant M15 protein. The extent of carboxymethylation, the effects on biological activity, antibody activation, physical stability, and the labeling of particular residues were studied. The results distinguish three groups of spatial relationships for cysteine residues in the protein, define possible regions for subunit interactions, and confirm that no cysteine residue is specifically involved in catalysis. Residue 1019 and to a lesser extent 498 are accessible in the tetrameric protein and probably represent exposed areas. In the M15 protein, these two, and three additional residues, at 76,387 and 600, were found to react significantly with reagent. One or more of the latter are suggested to be in the dimer-dimer interface. Complementation and activation by antibody are inhibited by carboxymethylation of M15 protein.


Temporal relationship of translation and glycosylation of immunoglobulin heavy and light chains.

The initial glycosylation of MPC 11 gamma 2b heavy chains occurs quantitatively in vivo when the nascent heavy chains reach a size of approximately 38 000 daltons. Nonglycosylated, completed MPC 11 heavy chains cannot be glycosylated in these cells. Other classes of mouse heavy chains (i.e., mu, alpha, and gamma 1) also appear to be glycosylated as nascent chains; nonglycosylated, completed heavy chains cannot be glycosylated by the cell in any of these cases. In contrast, variant MPC 11 cells synthesizing a heavy chain with a carboxy-terminal deletion appear to glycosylate some heavy chains prior to chain completion and some heavy chains after chain completion and release from the polysomes. Similar to the variant MPC 11 cells, MOPC 46B cells (which synthesize a kappa light chain containing an oligosaccharide attached to an asparagine located 28 residues from the amino terminus) glycosylate the majority of light chains after prior to chain completion but also some light chains after chain completion and release from the polysomes. In addition, it appears that, although completed MOPC 46B light chains can be glycosylated if they are present in a monomeric form, they cannot be glycosylated if they are present in a covalent dimeric form.


Acyldiglucosyldiacylglycerol-containing lipoteichoic acid with a poly(3-O-galabiosyl-2-O-galactosyl-sn-glycero-1-phosphate) chain from Streptococcus lactis Kiel 42172.

The lipoteichoic acid of Streptococcus lactis Kiel 42172 was isolated. The lipid portions were released by HF and were established to be 3-O-[O-alpha-D-galactopyranosyl-(1 leads to 6)-alpha-D-galactopyranosyl]-2-O-alpha-D-galactopyranosyl-sn-glycero-1-phosphate, they are joined by phosphodiester bonds nosyl)]glycerol. The repeating units of the hydrophilic chain were established to be 3-O-[O-alpha-D-galactopyranosyl-(1 leads to 6)-alpha-D-galactopyranosyl]-2-O-alpha-D-galactopyranosyl-sn-glycero-1-phosphate; they are joined by phosphodiester bonds at carbon atom 6 of the galabiosyl residues. The innermost unit is linked to the glycolipid by a phosphodiester presumably at C-6 of the outer glucosyl moiety. The hydrophilic chain is 7.4--11.8 units in length, measuring 12--19 nm is extended conformation. The content of 2.7--2.96 acyl groups per 2 glucosyl residues indicates that 70--96% of the glycolipid consists of acyldiglucosyldiacylglycerol. The novel poly(glycosylgly-cerophosphate) structure provided for the first time the oplipoteichoic acids are the sn-1 isomer which has previously been suggested from biosynthetic studies (Glaser, L., & Lindsay, B. (1974) Biochem. Biophys. Res. Commun. 59, 1131--1136).


k Chain variable regions from three galactan binding myeloma proteins.

A series of seven BALB/c myeloma proteins has been identified with binding specificity for antigens containing beta(1 leads to 6)-D-galactopyranosyl moieties. We have determined the primary amino acid sequence of the first 108 residues from the light chains of three of these proteins. The framework portions of the variable regions of these three light chains are identical with residue 100 at which position three different amino acids are found in the three chains. An additional interchange was found at position 106 in one of the proteins. Based on recent DNA sequence studies suggesting that the variable region ends at residue 97, these substitutions indicate the possible existance of multiple genes coding for the region beginning at residue 98 and continuing toward the carboxy terminus. A single amino acid interchange was observed in complementarity determining regions occurring in L3. This substitution (Ile-Trp) would require changes in all three codon bases to produce the respective amino acids if one were derived from the other. Two of these chains are thus indistinguishable for their first 100 amino acids and are the first pair of k chains to exhibit complete identity over their variable regions.


Human alpha-1-antichymotrypsin: purification and properties.

Human alpha-1-antichymotrypsin has been purified to homogeneity by the following sequential steps--(a) ammonium sulfate fractionation; (b) chromatography on Cibacron Blue Sepharose at pH 7.0; and (c) chromatography on SP-Sephadex C-50 at pH 5.5. The inhibitor has a molecular weight near 68,000 and contains approximately 26% carbohydrate alpha-1-Antichymotrypsin has an amino-terminal arginine and a carboxy-terminal glycine. It also has some homology with alpha-1-PI based on amino-terminal sequence analysis of both proteins. Complexes of alpha-1-antichymotrypsin with human chymotrypsin and human leukocyte cathepsin G are stable in sodium dodecyl sulfate and have molecular weights near 90,000 suggesting 1:1 complex formation on a molar basis between inhibitor and enzyme.


Involvement of the mature domain in the in vitro maturation of Bacillus subtilis precursor 5S ribosomal RNA.

A precursor of 5S ribosomal RNA from Bacillus subtilis (p5A rRNA, 179 nucleotides in length) is cleaved by RNase M5, a specific maturation endonuclease which releases the mature 5S rRNA (m5, 116 nucleotides) and precursor fragments derived from the 5' (21 nucleotides) and 3' (42 nucleotides) termini of p5A rRNA. Previous results (Meyhack, B., et al. (1978) Proc. Natl. Acad. Sci. U.S.A. 75, 3045) led to the conclusion that recognition elements in potential RNase M5 substrates mainly reside in the mature moiety of the precursor. Limited digestion of p5A rRNA with RNase T1 permitted the isolation of a number of test substrates which contained both precursor-specific segments and were unaltered in the immediate vicinity of the cleavage sites, but which differed in that more or less extensive regions of the mature moiety of the p5A rRNA were deleted. Tests of the capacity of these partial molecules to serve as substrates for RNase M5 indicate clearly that the enzyme recognizes the overall conformation of potential substrates, neglecting only the double-helical "prokaryotic loop" (Fox, G.E., & Woese, C.R. (1975) Nature (London) 256, 505).


Isolation and characterization of a pulmonary glycoprotein from human amniotic fluid.

A glycoprotein of the molecular weight of 36 000 has been isolated from human amniotic fluid. The glycoprotein was found to contain sialic acid, galactose, mannose, fucose, glucosamine, hydroxyproline and relatively high amounts of glycine. End-group analyses resulted in a single NH2-terminal residue indicating that the glycoprotein was homogeneous. The data indicate that this unique collagen-like glycoprotein, which is immunologically identical to a major alveolar glycoprotein found in alveoli of patients with alveolar proteinosis, is also a major protein in the human amniotic fluid. The idea that the pulmonary constituents enter the amniotic fluid cavity during fetal lung development is also confirmed by this report.


Studies on the characterization of the Rho(D) antigen.

The Rho(D) antigen of red cell membranes was solubilized using ethylene-diamine tetraacetic acid (EDTA) and 2-mercaptoethanol. The solubilized antigen was partially separated from other solubilized membrane components using molecular filtration. The antigen was treated with various enzymes to learn some of the chemical characteristics. It was found that the activity of the antigen, as measured by hemagglutination inhibition, was not affected by bee venom phospholipase A, Clostridium welchii phospholipase C, calf-intestinal alkaline phosphatase, Vibrio cholerae neuraminidase, pig kidney leucine aminopeptidase, bovine pancreatic carboxypeptidase A, and pig pancreatic carboxypeptidase B. However, the proteolytic enzymes, pronase, trypsin, chymotrypsin and papain, did destroy Rho(D) activity as measured by hemagglutination inhibition. These results indicate that protein is an important part of the active determinant of the Rho(D) antigen. The experiments by other investigators have shown that lipid is important to maintain the Rho(D) activity in the intact membrane; lipid probably helps to maintain the structural conformation of the Rho(D) molecule in its natural environment. The solubilized Rho(D) molecules are apparently not dependent on lipid for their Rho(D) activity.


Insulin counteraction of alpha-adrenergic effects on liver glycogen metabolism.

Insulin counteracted the effects of a pure alpha-adrenergic agonist, phenylephrine, on hepatocyte glycogen synthase and phosphorylase. These results argue against current concepts of insulin increasing cytoplasmic Ca2+ concentration.


Pinocytosis in the rat visceral yolk sac. Effects of temperature, metabolic inhibitors and some other modifiers.

Low temperature,2,4-dinitrophenol and moniodoacetate could each completely abolish the pinocytic uptake of 125I-labelled polyvinylpyrrolidone, 125I-labelled bovine serum albumin or colloidal 198 Au by 17.5-day rat visceral yolk sac cultured in vitro. Cytochalasin B and colchicine caused a partial and dose-dependent inhibition. It is concluded that the mechanism of pinocytic uptake of these substrates is not micropinocytosis as conventionally defined. Removal of extracellular calcium or the presence of theophylline inhibited liquid-phase pinocytosis by the rat yolk sac, whereas addition of ouabain caused a biphasic response: a slight stimulation of pinosome formation at a low concentration, and an inhibitory effect at a higher concentration.


Oxonol dyes as monitors of membrane potential. Their behavior in photosynthetic bacteria.

The reponses of oxonol dyes to single and multiple single turnovers of the photosynthetic apparatus of photosynthetic bacteria have been studied, and compared with the responses of the endogenous carotenoid pigments. The absorbance changes of the oxonols can be conveniently measured at 587 nm, because this is an isosbestic point in the 'light-minus-dark' difference spectrum of the chromatophores. The oxonols appear to respond to the light-induced 'energization' by shifting their absorption maxima. In the presence of K+, valinomycin abolished and nigericin enhanced such shifts, suggesting that the dyes, respond to the light-induced membrane potential. Since the dyes are anions at neutral pH values, they probably distribute across the membrane in accordance with the potential, which is positive inside the chromatophores. The accumulation of dye, which is indicated by a decrease in the carotenoid bandshift, poises the dye-membrane equilibrium in favor of increased dye binding and this might be the cause of the spectral shift. The dye response has an apparent second-order rate constant of approx. 2 . 10(6) M-1 . s-1 and so is always slower than the carotenoid bandshift. Thus the dyes cannot be used to monitor membrane potential on submillisecond timescales. Nevertheless, on a timescale of seconds the logarithm of the absorbance change at 587 nm is linear with respect to the membrane potential calibrated with the carotenoid bandshift. This suggests that under appropriate conditions the dyes can be used with confidence as indicators of membrane potential in energy-transducing membranes that do not possess intrinsic probes of potential.


Effects of lipid-phase separation on the filipin action on membranes of ergosterol-replaced Tetrahymena cells, as studied by freeze-fracture electron microscopy.

The effects of lipid-phase separation on the filipin action on pellicle membranes of ergosterol-replaced Tetrahymena pyriformis cells were studied by freeze-fracture electron microscopy. The pellicle membranes with phase separations induced by chilling from 34 degrees C (growth temperature) to lower temperatures (30, 22 and 15 degrees C) were treated with filipin. This produced filipin-induced lesions ("pits") only in the particulated (liquid) regions along the margin between solid and liquid domains, while they were produced in the particle-free (solid) areas when membranes were chilled to 15 degrees C. The pellicle membranes with lesions induced by filipin at 34 degrees C were chilled to 22 degrees C. This chilling raised larger particle-free areas and more condensed particle-aggregations on the membranes than on the membranes without the filipin treatment. These results suggest that the membrane fluidity affects induction and development of the ergosterol-filipin complex in the membrane.


Interaction between NADPH-cytochrome P-450 reductase and cytochrome P-450 in the membrane of phosphatidylcholine vesicles.

Cytochrome P-450 and NADPH-cytochrome P-450 REDUctase, both purified from liver microsomes of phenobarbital-pretreated rabbits, have been incorporated into the membrane of phosphoaditylcholine vesicles by the cholate dialysis method. The reduction of cytochrome P-450 by NADPH in this system is biphasic, consisting of two first-order reactions. The rate constant of the fast phase, in which 80--90% of the total cytochrome is reduced, increases as the molar ratio of the reductase to the cytochrome is increased at a fixed ratio of the cytochrome to phosphatidylcholine, suggesting that the rate-limiting step of the fast phase is the interaction between the reductase and the cytochrome. The rate constant of the fast phase also increases when the amount of phosphatidylcholine, relative to those of the two proteins, is decreased. This latter observation suggests that the interaction between the two proteins is effected by their random collision caused by their lateral mobilities on the plane of the membrane of phosphatidylcholine vesicles. The rate constant of the slow phase as well as the fraction of cytochrome P-450 reducible in the slow phase, on the other hand, remains essentially constant even upon alteration in the ratio of the reductase to the cytochrome or in that of the two proteins to phosphatidylcholine. No satisfactory explanation is as yet available for the cause of the slow-phase reduction of cytochrome P-450. The overall activity of benzphetamine N-demethylation catalyzed by the reconstituted vesicles responds to changes in the composition of the sysTEM IN A SIMILAR WAY TO THE FAST-PHASE REDUCTION OF CYTOCHROME P-450, though the latter is not the rate-limiting step of the overall reaction.


The mechanism of object fixation and its relation to spontaneous pattern preferences in Drosophila melanogaster.

The orientation behavior of walking flies, Drosophila melanogaster, towards a single 6 degrees wide black vertical stripe (elementary stripe) can be explained by use of the turning tendency function H(beta). This function is characterized by maximal values at an angular distance of beta = 25 degrees from the stable zero position (= orienting direction), a sharp decline from this maximum to beta = 60 degrees, and a very slow approach to the unstable zero position (Horn and Wehner, 1975). The shape of this function is influenced by both translatory and rotatory components of movement. If the translatory component is minimized by measuring the turning function W(beta) (see 2.3) at a distance of 10 mm (C1) from the center of the arena, a change in the strength of this decline is caused. But with increasing translatory component, i.e. at a greater distance from the center of the arena, W(beta) approximates the heuristical function H(beta) (Fig. 12). The turning functions W(beta) are pattern-specific; the angular positions of the maximum responses shift to greater angles with increasing width of the patterns (Fig. 2). In the two-pattern configuration with double or single stripes, there is always a coincidence between the stable zero positions of Wsigma(beta), the mean of the frequency distributions P(beta) of the flies' positions and ng(beta) of the straight courses, and the stable zero positions of Hsigma(beta) obtained from an additive superposition of two or more angular shifted turning tendency functions H(beta) (Fig. 5, 7). Therefore, the mean positions of the flies in a multi-stripe experiment composed of elementary stripes can be predicted from the addition of many angular shifted turning tendency functions H(beta). Between H(beta) and the frequency distribution P(beta) of the flies' positions beta, the following formula holds: P(beta) = C.intregal of H(beta)dbeta (Fig. 13). With this equation, the spontaneous preference of the broader of two double stripes can be explained presuming lateral interactions between the components of the patterns (Fig. 8, 10). The strength chii of this lateral interaction depends on the width delta of the double stripes. The greater delta, the smaller is chii. chii is a pattern-specific value (Table 1, 2).


[Ultrastructural changes in the cerebral cortex following transcranial micropolarization].

Electron microscopy of the cerebral cortex in cats and monkeys following transcranial micropolarization (TCMP) demonstrated ultrastructural changes whose degree was dependent on the electric current intensity and the stimulation period. In the focus of stimulation the current affected the brain tissue directly, different elements of the cerebral cortex showing unegual sensitivity to various TCMP regimens. The glia was the first to respond, then the neuronal bodies, and the last -- the synaptic structures. In the areas distant from the TCMP focus synaptic components altered first. The ultrastructural changes revealed were not of pathological character.


[Study of the "continuous cell antigen" and the human gastric mucosa].

The antigen common for continuous epithelial cell lines and gastric mucosa of humans described earlier was studied. This antigen was revealed in one more cell line, namely in that prepared from human mammary carcinoma MDA-MB-231, noncontaminated with HeLa cells. The antigen described can be detected in the exophytely growing adenocarcinomas of the stomach and in the mucosa of the carcinoma affected stomach at a distance of 10--12 cm from the site of affection; no such antigen was revealed in the endophytely growing carcinoma of the stomach and in mucosa areas surrounding gastric ulcer. The antigen is not a glycoprotein since glycoprotein fractions obtained by means of 1.2 M perchloric acid from the normal stomach mucosa homogenate and the E 16b extract were inactive in immunodiffusion with a sensitive serum. The electrophoretic mobility of the antigen was similar to that of globulin alpha1-beta2. This antigen is of interest since its detection or absence would possibly aid in determination of the initial type of cells from which development of carcinoma occurred, and in more precise recognition of the histological form of carcinoma of the stomach.


Afferent and efferent subcortical projections of behaviorally defined sectors of prefrontal granular cortex.

Although the functional significance of the midprincipalis region is well known, the afferent and efferent connections of this zone, in comparison to the anterior and posterior portions of the cortex lining the principal sulcus, are poorly understood. In 3 animals the retrograde tracer HRP and the anterograde tracers, tritiated proline, lysine and leucine, were injected into the sulcal cortex lining the principal sulcus. The cortex forming the banks of the principal sulcus was divided into anterior, middle and posterior sectors with one animal used for each zone. As expected from previous studies, the heaviest afferents to the cortex forming the principal sulcus were from the parvocellular portions of the medical dorsal nucleus. The medial pulvinar nucleus and the nucleus limitans projected to only the anterior and posterior portions of the cortex lining the principal sulcus. Projections were seen to all 3 sectors from the anterior, midline, intralaminar and lateral thalamic nuclei. Although cells were seen in the hypothalamus following injections in all 3 sectors of the cortex lining the principal sulcus, the heaviest hypothalamic projections were noted after injections into the mid-sector of the cortex. These HRP-positive cells were in the dorsal and lateral hypothalamic area, dorsal medial nucleus and in the lateral mammillary nucleus. These findings link the midprincipalis region with the prefrontolimbic circuit, and suggest that the midprincipalis region, n. medialis dorsalis, the mammillary bodies and perhaps the cingulate gyrus constitute part of an anatomical circuit concerned with memory processes.


Single-unit activity in the auditory cortex of monkeys selectively attending left vs. right ear stimuli.

Single-unit responses were recorded from the auditory cortex of rhesus monkeys that were performing an auditory selective attention task. Acoustic stimuli were presented randomly to either the left or right ear through headphones. In a given block of trials one ear was selected as the ear to be attended, and was indicated by lighting either a left or right response key. The animal's task was to press the lighted key whenever stimuli were presented to the attended ear, but to make no response to stimuli presented to the other ear. The attended ear was alternated on successive blocks of trials. Fourteen of 77 units showed significantly greater rates of evoked discharges for an attended stimulus than for an identical non-attended stimulus. The increase in stimulus-evoked activity was not accompanied by any increase in spontaneous activity or by any changes in the pattern of a unit's response. Changes in firing rate occurred at latencies as early as 20 msec.


Restricted posterior parietal lesions in the rhesus monkey and performance on visuospatial tasks.

Monkeys with lesions restricted to the inferior parietal lobule or the banks and depths of the superior temporal sulcus were tested on a route-following task. These areas are considered on neuroanatomical grounds to be homologous to parts of the human posterior parietal cortex, where lesions produce profound spatial disorientation. The operated monkeys were impaired on the route task, thus confirming at the behavioural level the anatomical predictions of comparability between parietal cortex in monkey and that in man. The monkeys were not impaired, however, on a visual pattern discrimination or on a visual-spatial task with cue and response separation, a task that was considered on the basis of previous investigations with extensive posterior lesions in the monkey to be sensitive to parietal lesions.


Response of human lymphocytes to mitogen: at what stage is there a requirement for Ca2+?

In order for significant DNA synthesis to be observed in the case of human lymphocytes stimulated for 36 h in presence of phytohemagglutinin (PHA), Ca2+ must be present in the medium continuously for at least 20 h; access to Ca2+ for 10-h periods during the first 30 h was not sufficient to permit DNA synthesis to occur. Addition of the chelator EGTA from 0 to 60 h after stimulation caused severe inhibition of incorporation of labelled thymidine when this was measured after 36 to 144 h of culture. Equimolar calcium reversed the inhibition caused by EGTA. Incorporation of labelled uridine and leucine showed a temporal pattern of dependence on the presence of Ca2+ in the medium similar to that of thymidine. Ca2+ appears not to be required in the medium during the last half (i.e., 20-36 h) of the presynthetic G1 phase nor during S phase since removal of Ca2+ from the medium after 20 h did not prevent a subpopulation of lymphocytes from entering S phase 16 h later.


Chromosome age and segregation during sporulation of Bacillus megaterium.

The effect of chromosome age on segregation during sporulation was investigated. Vegetative cells of Bacillus megaterium were labeled with [Me-3H]thymine and then were grown at 30 degrees C in nonradioactive medium for various times before being allowed to sporulate. The ratio of the amount of label in sporal DNA to that in sporangial DNA, obtained after minor correction for the sporulation frequency, remained essentially constant as the postlabeling growth period was increased from one to seven generations. The spores were preferentially located at the older poles of sporangia, i.e. the poles formed by divisions occurring prior to those forming the sporangia. Therefore, it seems that old (labeled) chromosomes segregate randomly with respect to both the morphological and genealogical polarities of sporangia. Examination of total cell lysates by dye-buoyant density gradient centrifugation revealed the presence of covalently closed circular DNA from cells grown at 37 degrees C, but none was obtained from cells grown at 30 degrees C. Thus, possible interference by large amounts of extrachromosomal DNA in the determination of the chromosomal segregation pattern is unlikely.


Isolation of tuberculin peptides from tuberculin purified protein derivative (PPD).

Tuberculin purified protein derivative (PPD) obtained from the filtrate of Mycobacterium tuberculosis was hydrolysed with proteinase, trypsin, or chymotrypsin. Each hydrolysate consisted of a tuberculin peptides mixture (TPM). From each TPM 16 fractions were obtained by ion-exchange chromatography on Dowex 50W-X8 but only one fraction was isolated from each of the 16 fractions which showed tuberculin activity in guinea pigs sensitized with M. bovis (bcg) or M. tuberculosis. This fraction was designated "purified tuberculin peptide" (PTP). The PTP fraction from the proteinase hydrolysate (PTP-proteinase) was rechromatographed on Dowex 1-X2 and two tuberculin peptide fractions having molecular weights of 3200 and 12,000 were isolated. The potency of these two fractions was assessed in guinea pigs sensitized with M. bovis (BCG) and with M. tuberculosis and they were approximately 4 to 7 times more potent than either the international standaCG and of at least equal potency to either PPD-S or Connaught PPD in guinea pigs sensitized with either M. kansasii, M. scrofulaceum, M. intracellulare, or M. avium whereas very little if any cross-reactivity was elicited by these two fractions. This lack of response indicates that either fraction could be used as an aid to differentiate between sensitization due to M. tuberculosis or M. bovis and sensitization attributed to other mycobacteria.


An evaluation of fast neutron beam teletherapy of metastatic cervical adenopathy from squamous cell carcinomas of the head and neck region.

One hundred thirteen patients with metastatic cervical adenopathy (MCA) from advanced squamous cell carcinoma of the head and neck completed fast neutron beam irradiation between September 1976 and March 1977. With a maximum follow-up time of 49.9 months (mean 14.5 months), 54% of the total group remain disease-free at the sites of their cervical adenopathy. Control of adenopathy measuring 3 cm or less in maximum dimension was 75% with neutrons only and 87.5% with mixed beams. Nodes measuring 3-6 cm were controlled 38.1% of the time with neutrons only and 59% of the time with mixed beams. Clinical control of adenopathy measuring greater than 6 cm was obtained in 12.5% with neutrons only and 29.4% with mixed beams. For patients with controlled primary tumors, the overall control rate in the neck was 72%; 100% of nodes 3 cm or less were controlled, 82% of nodes measuring from 3-6 cm were controlled, and 42% of nodes greater than 6 cm were controlled. Patients with controlled primary tumors treated with mixed beams had 100% control of nodes measuring 3 cm or less, 96% control of nodes measuring 3-6 cm, and 50% control of nodes measuring greater than 6 cm. Overall, the control rate for mixed beams was significantly better than for treatment with neutrons only. Morbidity was less (both acute and late) in the mixed beam-irradiated group.


Vinblastine, adriamycin, thiotepa, and halotestin (VATH): therapy for advanced breast cancer refractory to prior chemotherapy.

Nineteen postmenopausal patients with metastatic breast cancer refractory to conventional combination chemotherapy were treated with monthly cycles with the combinations of vinblastine, adriamycin, thiotepa and halotestin. Ten patients (52%) responded with a greater than 50% regression of measurable tumor. The median duration of response was 11.5 months, with 5/10 patients still responding at a mean follow-up of 10 months. Only 2/10 responders have died with a mean follow-up of 13.8 months. In contrast, 8/9 nonresponders have died (median survival 6.0 months). Response to therapy was neither influenced by site of disease, time interval from diagnosis to primary chemotherapy nor duration of response to primary chemotherapy. No patient was hospitalized because of drug induced toxicity. This combination of drugs is a tolerable effective regimen for patients relapsing after adjuvant chemotherapy or after primary combination chemotherapy for grossly metastatic disease.


Limited epithelial carcinoma of the ovary treated with curative intent by the intraperitoneal installation of radiocolloids.

Between January 1960 and September 1972, 104 patients with limited epithelial carcinoma of the ovary received intraperitoneal radiocolloid. Fifty-six of these patients also received external beam radiation therapy to the pelvis (pelvic RT). Five-year actuarial no-evidence-of-disease survival rates were 95% for stage Iai, 82% for Iaii, 73% for Ib, 67% for Ic, 67% for IIa, 67% for IIb without gross residual tumor (GRT), 25% for IIb with GRT, and 50% for III with minimal or no GRT. The addition of pelvic RT following radiocolloid could not be shown to affect survival of patients with Stage I and IIa tumors. Small bowel complications were related to the use of pelvic RT, however, occurring in 2.2% of patients treated with radiocolloid alone and 24% of patients treated with colloid and pelvic RT (p less than 0.005). In patients who underwent abdominal surgery following treatment of ovarian cancer, no excessive complication rate was observed. We conclude that for patients with stages Iaii through IIa, postoperative radiocolloid appears to provide the greatest chance of survival with the least chance of complication. For patients with Stage IIb and III lesions in whom there is minimal or no GRT, radiocolloid followed by pelvic RT produced survival rates comparable or superior to any other form of postoperative therapy.


Effect of Cobalt-60 radiation on response to endodontic therapy in monkeys.

Response of teeth that had received therapeutic doses of Cobalt-60 radiation to endodontic therapy were investigated in three monkeys. The results indicated no appreciable effect of the irradiation on the response to root canal treatment aside from reduction in osteoblastic activity.


Juvenile ossifying fibroma: an ultrastructural study.

The ultrastructural features of a juvenile ossifying fibroma of the maxilla are described. The stromal portion of the tumor was composed of osteoblasts and to a lesser extent of fibroblasts. The bone spicules were rimmed by osteoblasts and osteoclasts. Calcification was seen to occur along the collagen fiber matrix, corresponding to calcification of osteoid, and also in the form of intracellular and extracellular crystallization. The latter form of calcification corresponded to so-called psammoma-like bodies, and was considered characteristic of this subtype of ossifying fibroma.


Cryocrystalglobulinemia in hairy cell leukemia.

A patient with hairy cell leukemia (leukemic reticuloendotheliosis) was noted to have a spurious leukocytosis caused by a spontaneously crystallizing cryoglobulin. The cryoprotein was identified as IgG lambda. An intracytoplasmic immunoglobulin demonstrable within the hairy cell was also IgG lambda. The cryoglobulin spontaneously disappeared over a four day period. A reliable automated count on the Coulter Model S could be obtained by prewarming the blood specimen to 37 C.


Correlation of UDP-galactose glycoprotein:galactosyltransferase levels in the sera with the clinical status of ovarian cancer patients.

Uridine diphosphate-galactose : glycoprotein galactosyltransferase (EC 2.4.1.22) was measured serially prior to and after surgery in 4 patients with ovarian epithelial cancer. The levels of this enzyme in the sera correlated well with the clinical status of the patients. In 2 other patients, the follow-up was designed to detect recurrence, and the enzyme assay was started when the patients were clinically disease free. Elevation of galactosyltransferase preceded the clinical appearance of disease by 3-7 months. Serial determination of glycoprotein galactosyltransferase in serum may be useful for evaluating the effectiveness of therapeutic programs.


Pharmacologic factors and manipulation of immunity systemic adjuvants in cancer therapy.

Because of the experimental and clinical studies which have been extensively conducted with bacillus Calmette-Guérin (BCG) as a systemic adjuvant in cancer immunotherapy, we have analyzed the main factors and conditions which determine its beneficial action and have underlined some of these (eg, the dose factor which controls the amplification of suppressor cells which is probably responsible for failures and even the possible tumor-enhancing effect of immunotherapy). Knowing those factors and conditions, we have been able to establish a systematic immunopharmacologic study of systemic immunity adjuvants, which has resulted in the discovery of agents whose actions are more rapid than that of BCG on one or a few populations of cells involved in immunity and which, unlike BCG, do not induce suppressor cell amplification. This amplification may explain the difference in the results obtained with this mycobacterium in various clinical immunotherapy trials in which it was applied differently. It is proposed to combine these mono- or pauc-functional adjuvants in order to try to obtain all of the beneficial effects of BCG without the amplification of suppressor cells.


Effect of levamisole on metabolism of phagocytic cells.

The effect of levamisole (LMS) on glucose metabolism was studied using a protozoan phagocytic model and human leukocytes. At concentrations of greater than 10 microgram/ml, LMS inhibited glucose metabolism in the protozoan phagocytic model. Glucose metabolism in both the phagocytic model and normal peripheral blood leukocytes was, however, minimally altered when exposed to levels of LMS of less than 10 microgram/ml. In contrast, myeloblasts from a child with leukemia seemed to have increased metabolic activity and 5 microgram/ml of LMS markedly reduced the glucose metabolism. These preliminary studies suggest that LMS can alter glucose metabolism of certain cells and that some malignant cells may be directly inhibited metabolically by LMS.


Relationship of molecular weight to antiviral and antitumor activities and toxic effects of maleic anhydride-divinyl ether (MVE) polyanions.

The molecular weight (MW) and dose dependency of several of the toxic effects and antitumor and antiviral activities of a new series of five maleic anhydride-divinyl ether copolymers (MVE) were established. Each polyanion preparation was relatively homogeneous and exhibited a narrow MW range, from 12,500 (MVE-1) to greater than 52,000 (MVE-5). All of the polyanions were effective as adjuvants to surgery against the metastatic Lewis lung carcinoma, and also exhibited marked antitumor activity against the P815 mastocytoma. MVE-1 retained antitumor activity while losing considerable antiviral activity. This polyanion also exhibited the least toxicity with regard to criteria such as sensitization to the lethal effects of endotoxin, inhibition of reticuloendothelial function, and depression of the microsomal mixed functional oxidase system. The MVE-4 (MW, 32,000) and MVE-5 (MW, 52,600) polyanions exhibited potent antitumor and antiviral activity, but also demonstrated dose-dependent toxic effects.


Macrophage activation and antitumor activity of a Brucella abortus ether extract, Bru-Pel.

Bru-Pel and Brucella abortus lipopolysaccharide (LPS) were tested for both macrophage activation and antitumor activity in an artificial metastasis model. Resting macrophages were rendered nonspecifically tumoricidal for MBL-2 lymphoblastic leukemia target cells by exposure to Bru-Pel at greater than or equal to 1 ng/ml of culture medium. B. abortus LPS failed to activate macrophages in vitro at all concentrations tested. Ip treatment of homozygous nude mice with Bru-Pel induced cytotoxic macrophages, indicating that Bru-Pel activated macrophages through a thymic-independent process. An artificial metastasis model was developed where single-cell suspensions of Madison 109 lung carcinoma were inoculated iv into syngeneic BALB/c mice. Bru-Pel, but not B. abortus LPS, strikingly inhibited tumor-colony formation in the lungs. Although Bru-Pel contains endotoxin, the data demonstrate that endotoxin is apparently not the active component by which Bru-Pel activates macrophages and enhances host resistance to cancer.


Ultastructural localization of polysaccharides in the vacuolar system of an established cell line.

Polysaccharides in the vacuolar system of an established line of monkey kidney epithelial cells, were investigated by high resolution radioautography following incorporation of 3H-fucose and by the periodic acid-thiosemicarbazide-silver proteinate staining method. Significant amounts of glycogen and glycoproteins were found in different lysosomes and prelysosomes. The radioautographic labeling of these substances correlated with the cytochemical observations. Analysis of the 3H-fucose pattern of labeling suggests that glycoproteins are transported through the different components of the vacuolar system of the cell in a sequential fashion rather than via independent pathways. The possible functional significance of polysaccharides in lysosomes is discussed. It is suggested that glycogen is taken up by lysosomes through autophagic segregation and through infolding and vesiculation of the lysosomal surface, and that the majority of glycoprotein in the lysosomal membranes does not have acid hydrolase activity.


Fine structural and cytochemical study of the innervation of smooth muscle in an amphibian (Bufo marinus) lung before and after denervation.

The innervation of the toad (Bufo marinus) lung was studied with transmission electron microscopy and fluorescence techniques, both before and after 12 or 20 days close vagosympathetic denervation. Four cytologically distinct types of neuronal processes were recognised, in relation to the visceral muscles of the lung. These were described as cholinergic, adrenergic, non-adrenergic/non-cholinergic (NANC) and sensory on the basis of the characteristics of their vesicular content and cytochemical reactions. An apparent efferent innervation of visceral smooth muscle was achieved by NANC (50%), cholinergic (25%) and adrenergic (25%) fibres. A few sensory fibres were also present. After denervation only NANC fibres persisted, showing that the cell bodies of these fibres were intrapulmonary. The vascular smooth muscle was supplied by cholinergic, adrenergic and sensory fibres. In the walls of the proximal branches of the pulmonary artery were fibres containing large dense-cored vesicles. These profiles, which were associated with the vasa vasorum, were similar to neurosecretory fibres. After denervation all neural profiles associated with the vasculature had degenerated. The observations suggest that vagal vasodepressor effects in the toad lung are mediated indirectly through relaxation of visceral muscle strands which in their contracted state compress vascular channels.


Organization of the sympathetic innervation in liver tissue from monkey and man.

The sympathetic innervation of the liver of monkey and man has been investigated in a combined fluorescence histochemical, chemical and electron microscopical study. By means of the Falck-Hillarp fluorescence method a dense network of monoamine-containing nerve fibers was visualized in liver tissue of monkey and man. The nerve fibers ran in close contact to both hepatocytes and blood vessels. Chemical quantitations showed high concentrations of noradrenaline in both human and monkey liver. Microspectrofluorometry of the intraneuronal monoamine resulted in spectra characteristic of  a catecholamine. For the electron microscopical study the dopamine analogue, 5-hydroxydopamine, was used to "label" the catecholamine terminals in both human and monkey liver. The nerve profiles, identified as catecholamine-containing, were demonstrated in a perivascular location and in close contact to hepatocytes. No synaptic membrane specializations were present between nerve fibers and hepatocytes. The general ultramorphology and intralobular distribution pattern of nerves in the liver of monkey and man were similar. The present results prove the existence of a sympathetic innervation of hepatocytes and blood vessels in the liver of man and monkey.


Innervation and cytochemistry of the neuroepithelial bodies in the ciliated epithelium of the toad lung (Bufo marinus).

Neuroepithelial bodies (NEB) were identified in the lung of Bufo marinus. The characteristics of the cells and their innervation were studied with electron and fluorescence microscopy before and after close vagosympathetic denervation. The bodies consist of low columnar cells which rest on the epithelial basal lamina. The majority of the cells do not reach the lumen of the lung (basal cells); the few which do (apical cells) are bordered by microvilli and possess a single cilium. The neuroepithelial cell cytoplasm contains a variety of organelles the most characteristic of which are dense cored vesicles. Microspectrofluorometry and electron microscopic cytochemistry indicate significant quantities of 5-hydroxytryptamine in these cells. The neuroepithelial bodies could be divided into three groups on the basis of their innervation: 1) About 60% of the NEBs are innervated solely by nerve fibers containing agranular vesicles which form reciprocal synapses; 2) about 20% are innervated solely by adrenergic nerve fibres which from distinct synaptic contacts; and 3) the remaining 20% are innervated by both types of nerve fibres. It is proposed that the NEBs are receptors monitoring intrapulmonary P CO2 and so leading to modulation of activity in afferent nerve fibres (type containing a granular vesicles). The presence of NEBs soley with an adrenergic (efferent) innervation poses a problem with this interpretation.


The innervation and fine structure of paraneuronic cells in an amphibian pulmonary artery.

The pulmonary artery of Bufo marinus contains large numbers of bipolar cells situated in the tunica adventitia and in the outer layers of the media. These cells show a bright green-yellow fluorescence (emission spectra 485 nm) after formaldehyde pre-treatment suggesting that they contain a primary monoamine. The most characteristic fine-structural feature of these cells is the presence of numerous dense-cored vesicles (80--300 nm diameter) in their cytopalsm. The cells are in close contact (20 nm gap) with both agranular and granular nerve fibres. Both EM-cytochemical and formaldehyde-induced fluorescence tests indicate that the granule-containing nerve fibres are adrenergic. The agranular nerve fibres form discrete synaptic contacts with pre and post-synaptic membrane thickenings on the cells. This was never observed with respect to the adrenergic fibres. Each process of the cells is about 45 micrometer long. The processes do not bear any special relationship to either vessels of the arterial vasa vasorum or medial smooth muscle cells. Their location in the wall of the artery suggests that they are functionally significant with respect to activity of the arterial media.


Complete sequence of constant and 3' noncoding regions of an immunoglobulin mRNA using the dideoxynucleotide method of RNA sequencing.

Three synthetic oligonucleotides were prepared to be complementary to known regions of the mouse immunoglublin light chain mRNA, and their ability to prime the transcription of complementary DNA (cDNA) was studied. The sequence of the cDNA was determined by adapting for mRNA the DNA sequencing method of Sanger, Nicklen and Coulson (1977) which uses 2'3' dideoxy ribonucleotides. A continuous sequence of 532 nucleotides was obtained, 321 corresponding to the whole of the constant region of the mRNA and the remaining 211 being the complete 3' noncoding region of the mRNA. The termination codon U-A-G occurs at the expected position in the mRNA corresponding to the triplet following the C terminal cystine. The nucleotide sequence is partially corroborated by the sequence of fragments obtained previously from 32P-mRNA fingerprints and endonuclease IV digests of 32P-cDNA, and is in agreement with the amino acid sequence of the constant region, except for a rearrangement of four amino acids (between amino acid positions 163 and 166). A revision of the amino acid sequence confirms the nucleic acid sequence.


Genetic and molecular organization of the 5S locus and mutants in D. melanogaster.

The topography of an entire redundant locus was analyzed by both genetic and molecular means. Three mutants (min0, min1, min2) allelic to the 5S rRNA genetic locus on chromosome 2 of D. melanogaster were isolated. Flies exhibit a mutant phenotype when hemizygous for a min allele, but flies having two doses are wild-type. Saturation hybridization experiments show that the alleles are gross defieciencies each deleting an equal amount of 5S DNA. Each of the three mutant min alleles produces a distinct temperature-sensitive viability phene, and thus they are suggested to be pseudoalleles within the same redundant locus. Using the segmental aneuploid method (Lindsley et al., 1972), the 5S gene cluster was subdivided into proximal and distal halves. Both saturation hybridization experiments and genetic tests show that each half contains about eighty 5S genes. The complementation of the min alleles with the proximal and distal halves of the cluster indicates that both halves function independently. We present evidence which supports the model that all of the 160 5S genes are arranged as a single continuous cluster of tandem repeats with no large interdispersive DNA segments not complementary to 5S rRNA.


One of the copia genes is adjacent to satellite DNA in Drosophila melanogaster.

A method for purifying sequences adjacent to satellite DNA in the heterochromatin of D. melanogaster is described. A cloned DNA segment containing part of a copia gene adjacent to 1.688 g/cm3 satellite DNA has been isolated. The copia genes compose a repeated gene family which codes for abundant cytoplasmic poly(a)-containing RNA (Young and Hogness, 1977; Finnegan et al., 1978). We have identified two major poly (A)-containing RNA species [5.2 and 2.1 kilobases (kb)] produced by the copia gene family. The cloned segment contains copia sequences homologous to the 5' end of RNA within 0.65 kb of the 1.688 satellite DNA sequences. Seven different cloned copia genes from elsewhere in the genome have also been isolated, and a 5.2 kb region present in five of the clones was identified as copia by heteroduplex analysis. In addition, three ususual copies of copia were found: a "partial" copy of the gene (3.7 kb) which has one endpoint in common with the 5.2 kb unit; a copia gene flanked on one side by a 1.6 kb sequence and on the other by the same 1.6 kb sequence in the inverted orientation; and a copia gene flanked only on one side by the same sequence.


Heat shock and phenocopy induction in Drosophila.

Differentiation of the single cells that yield bristles and hairs in Drosophila has been examined by studies of phenocopies induced by heat shock. Phenocopies that resemble the mutants hook and javelin are produced by application of the stress within narrow and specific time ranges during metamorphosis. Double phenocopies can be induced by successive heat shocks at appropriate time intervals. Phenocopy production may be related to heat shock effects on RNA synthesis through a feedback inhibition of transcription by heat shock proteins.


Sequences of mouse immunoglobulin light chain genes before and after somatic changes.

We have determined the nucleotide sequences of the germ line gene as well as a corresponding somatically mutated and rearranged gene coding for a mouse immunoglobulin lambdaI type light chain. These sequencing studies were carried out on three Eco RI-DNA fragments which had been cloned from BALB/c mouse embryos or a lambdaI chainsecreting myeloma, H2020. The embryonic DNA clone Ig 99lambda contains two protein-encoding segments, one for the majority of the hydrophobic leader (L) and the other for the rest of the leader and the variable (V) region of the lambda0 chain (Cohn et al., 1974); these segments are separated by a 93 base pair (bp) intervening sequence (I-small). The coding of the V region ends with His at residue 97. The second embryonic DNA clone Ig 25lambda includes a 39 bp DNA segment (J) coding for the rest of the conventionally defined V region (that is, up to residue 110), and also contains the sequence coding for the constant (C) region approximately 1250 untranslated bp (I-large) away from the J sequence. The J sequence is directly linked with the V-coding sequence in the myeloma DNA clone, Ig 303lambda, which has the various DNA segments arranged in the following order: 5' untranslated region, L, l-small, V linked with J, l-large, C, 3' untranslated sequence. The lg 303lambda V DNA sequence codes for the V region synthesized by the H2020 myeloma and is different from the lg 99lambda V DNA sequence by only two bases. No silent base change was observed between the two DNA clones for the entire sequence spanning the 5' untranslated regions and the V-coding segments. These results confirm the previously drawn conclusion that an active complete lambdaI gene arises by somatic recombination that takes place at the ends of the V-coding DNA segment and the J sequence. No sequence homology was observed at or near the sites of the recombination.


The synthesis and processing of the messenger RNAs specifying heavy and light chain immunoglobulins in MPC-11 cells.

The nuclear precursors of the immunoglobulin messenger RNAs of MPC-11 cells were characterized with respect to size, amount per cell and extent of polyadenylation. These cells produce three Ig mRNAs: a 1.8 kb component coding for a gamma2b heavy chain (H mRNA), a 1.2 kb mRNA coding for a k light chain (L mRNA) and a 0.8 kb mRNA coding for the constant region portion of the k light chain (Lf mRNA). To identify the pre-mRNAs without ambiguity, we constructed recombinant DNA plasmids containing H and L cDNA sequences, and used the cloned cDNAs as hybridization probes for analysis of steady state nuclear RNA and in DNA excess hybridization experiments with pulse-labeled nuclear RNA. The nuclear molecules containing Ig sequences consist of an 11 kb component (H1), which we believe to be the primary transcript of the H gene, 5.3 kb (L1), and 3.3 kb (L2) components, which seem to be primary transcripts of the L and L1 genes, components corresponding to mature size H, L and Lf mRNAs, and several intermediate-sized components which include the processing derivatives. The precursor role of these nuclear molecules was established by studies of their labeling kinetics and by appropriate pulse-chase experiments. All the pre-mRNA species including H1, L1 and L2 contain poly(A), thus suggesting that polyadenylation is an early event in the processing of these mRNAs. The MPC-11 cell contains about 30,000 and 40,000 cytoplasmic H and L mRNA molecules, respectively, which must be produced within one cell generation (approximately 24 hr). In comparison, the nucleus contains about 100-150 molecules of total pre-mRNA and only about 10-15 molecules of presumptive primary transcripts for each of these Ig species. These values indicate very rapid transcription rates (greater than 20 transcripts per min) and exceptionally fast processing rates (approximately 0.5 min for the primary transcripts and approximately 5 min for overall nuclear processing) for the Ig mRNAs. Thus rapid transcription and processing, together with high cytoplasmic stability, account for the high abundance of Ig mRNAs in the myeloma cell.


[Decrease in stimulation and allogeneic response following experimental infection of mice with human adenovirus 5].

15 days after experimental infection with human adenovirus 5, C57 Bl/6 murine spleen cells were found to have altered in vitro properties. Proliferative response to phytohemagglutinine was slightly increased while the allogeneic response was decreased and the capability to stimulate allogeneic cells was significantly diminished. The degree of expression of surface antigens responsible for allogeneic stimulation is influenced by many regulatory factors, still poorly defined; alteration of some factors of antigenicity may result, directly or indirectly, from certain viral infections.


[The existence in Europe of four groups of mice (genus Mus L.) of species and subspecies rank, demonstrated by biochemical genetics].

Analysis at 20 genetic loci by starch gel electrophoresis of a population from Greece shows that contrary to most accepted views, the outdoor Eastern Mediterranean Short-tailed Mouse (i) does not belong to the same species as the House Mouse (Mus musculus), (ii) is not less distant from the Eastern European semi-species of House Mouse (biochemical group 2) than from the Western and Mediterranean European semi-species (group 1), and (iii) is not more closely related to the outdoor Western Mediterranean Short-tailed Mouse Mus spretus (group 3), than to M. musculus. It represents a new biochemical group (group 4) and deserves a species rank (Mus spicilegus).


[Microspectrophotometric study of biogenic amines in ganglions of the central nervous system of Crepidula fornicata (Phil.)].

The fluorescent products formed from biogenic amines treated with formaldehyde according to the method of Falck and Owman have been studied in the brain of Crepidula fornicata. The microspectrophotometric analysis of cellular indolamines shows excitation emission maxima at 385--525 nm (serotonine) and catecholamine at 410--475 nm upon HCl treatment catecholamine products show a shift of the excitation peak from 410 to 370 nm (dopamine).


[Presence of substances immunoreactively related to enkephalin in an insect, Locusta migratoria].

Application of specific radioimmunoassays to various extracts of the locust showed that nervous tissues and specially the optic lobes contain enkephaline-like material. However these immunoreactive substances present an apparent higher molecular weight (700--1 000 dalton) than those of the controls, as tested by chromatography on Sephadex G 25.


[Molecular weight of lipovitellins during egg development in the crustacean amphipod Orchestia gammarella (Pallas)].

Egg lipovitellins of Orchestia gammarella tested by electrophoresis on gels of different acrylamide concentrations, following the procedure of Hedrick and Smith (1968), display a migration pattern identical to that of proteins with molecular weights of congruent to 3.2 x 10(5) (lipovitellin I) and congruent to 5.5 x 10(5) (lipovitellin II) respectively. Since their molecular weight remains constant during embryogenesis, the changes of their relative mobility in disc gels indicates alterations in their ionic charges.


[Initiation of germination of Clostridium difficile spores by lysozyme].

The germination rate of spores of C. difficile which is usually lower than 10(-5) is raised to about 5.10(-3) in presence of lysozyme. All spores are initiated by lysozyme when previously treated by sodium thioglycolate. These spores are indeed lysozyme-dependent for germination.


[Effect of lymphokines produced in a grafted system on production of prostaglandins by macrophages].

Mice peritoneal macrophages cultured in vitro release a prostaglandin-like activity as evaluated by a bio-assay using Rat stomach fundus. The prostaglandin release is greatly increased when macrophages are incubated with the supernatant of mixed lymphocyte cultures between recipient and donor of a skin allograft. This phenomenon was found in all strains of Mice tested except C3H/HeJ Mice, a strain already known for its defective responsiveness to bacterial lipopolysaccharide (LPS).


[The microvascularity of human placental villi].

Scanning electron microscopic observation of plastic casts of human chorionic villi vasculature has demonstrated that in the area of placental exchange two blood capillary networks are present: the paravascular and the villous capillary networks. S.E.M. provides precise information on the shape and anatomical relations of these blood capillary networks.


[Seasonal plasma testosterone variations in the stone marten].

Plasma testosterone levels change with annual sexual cycle of the stone marten. Very low during the decrease and the quiescence of the testis, it rises during the testis onset and reaches a maximal peak during the coït period (May to July).


[Absorption of double labeled (glycerol 3H-linoleic acid 14C) native bile by phosphatidylcholines].

Double-labeled bile ([U. 3H glycerol] [1. 14C linoleic acid])--in which about 70% of labeling 14C and 80% of labeling 3H of total lipids were borne by phosphatidylcholines (PC), (isotopic ratio of these PC was equal to 1)--was introduced into the duodenum of test Rats, some of them with a bile fistula. As low amounts of the hydrolysis products of biliary PC were found in the intestinal lumen, a higher hydrolysis must occur further (brush border, enterocyte ?) because, in the mucosa, the highest labeling 14C was present as triglycerides and PC have an isotopic ratio 3H/14C higher than 1. As in the lumen the isotopic ratio 3H/14C of PC was higher than 1 and increased with the time elapsed, this finding suggests that mucosal PC were added to biliary PC (secretion or desquamation ?) unless these modifications were due to luminal micro-organisms. As test Rat bile was poorly labeled a very weak enterohepatic circulation of biliary diunsaturated PC may exist.


[Nicotinamide desamidase in the culture medium of neuroblastoma cells].

Important amounts of nicotinic acid appear in the growth medium of neuroblastoma cell cultures. It is shown that an enzyme released by the cells into the growth medium deamidates the nicotinamide supplied by the growth medium to nicotinic acid.


[Effect of platinum derivatives on the inducible and repressible liver microsomal enzyme systems in the rat: inhibition of zoxazolamine-hydroxylase and induction of dimethyl-nitrosamine demethylase isoenzymes by cis-dichlorodiamine platinum (cis-PtCl2(NH3)2) and ammonium hexachloroplatinum (PtC16(NH4)2)].

Two platinum derivatives, cis-PtCl2(NH3)2 and PtCl6(NH4)2 have been studied for their effects on the Rat on cytochrome P450 in hepatic parenchyma on zoxazolamine-hydroxylase, a typical inducible system and on the two isoenzymes of dimethyl-nitrosamine demethylase, typical repressible systems. The inhibitory effect of PtCl6(NH4)2 on zoxazolamine-hydroxylase activity, previously shown by the authors, has been confirmed. The cis-PtCl2(NH3)2 also significantly inhibits zoxazolamine-hydroxylase activity. On the other hand, both of the platinum derivatives decrease cytochrome P450 level and enhance the dimethyl-nitrosamine metabolism. These various effects and their relationship are discussed.


[Epileptogenic action of intra-amygdaloid injection of kainic acid].

Intra-amygdaloid unilateral application of low doses of kainic acid rapidly elicits, in both chronic and acute conditions, secondarily generalized convulsive seizures which often culminate in fatal true status epilepticus unless appropriate anti-epileptic drugs are provided. Spontaneous recurrent seizures are observed for several hours starting approximately 10 min. after the application of kainic acid. In addition to the primary degeneration in the amygdaloid complex, a secondary cell loss is seen in CA3 area of the dorsal hippocampus. It is suggested that this procedure may constitute a particularly suitable model for the study of true focalized status epilepticus.


[Role of the infundibular complex and its neural afferents in the photosexual reflex in the quail].

The photosexual reflex was suppressed in quail after bilateral preoptic lesions. However an anterior-lateral de-afferentation of the hypothalamus of previously preoptic-lesioned quail resulted in partial photo-induced testis growth. As a working hypothesis, it is suggested that the preoptic area may control some unknown extrahypothalamic influences in order to modulate the gonadotropin controlling function of the infundibular area.


[Action of cadmium on a free dinoflagellate Prorocentrum micans E: growth, cadmium absorption and cell modifications].

The study of the growth of Prorocentrum micans E. submitted to sublethal doses of cadmium demonstrated a high sensitivity of this Dinoflagellate to the polluant starting with concentration of 5 microgram/l. The uptake of cadmium was measured by pulse polarographic analysis of the filtered culture medium. Structural modifications, not apparent in living cells, are visible after fixation. The high sensitivity of Prorocentrum micans to cadmium, also its easy culture, makes it a suitable pollution indicator of this heavy metal.


[Spatial conformation of human serotransferrin glycans].

The construction of molecular models for the human serotransferrin glycans shows that they present one compact section linked to the protein and constituted by the pentasaccharide alpha-Man-(1 leads to 3)-[alpha-Man-(1 leads to 6)]-beta-Man-(1 leads to 4)-beta-GlcNAc-(1 leads to 4)-beta-GlcNAc-(1 leads to)-Asn to which are attached two "antennae" consisting of the trisaccharide alpha-NANA-(2 leads to 6)-beta-Gal-(1 leads to 4)-beta-GlcNAc. The trisaccharide sequence beta-Man-(1 leads to 4)-beta-GlcNAc-(1 leads to 4)-beta-GlcNAc adopts a flat and rigid conformation, stabilised by hydrogen bonds. In contrast, the sequence alpha-NANA-(1 leads to 6)-beta-Gal-(1 leads to 4)-beta-GlcNAc-(1 leads to 2)-alpha-Man takes up a helical configuration. The two "antennae" can be disposed on the pentasaccharide core to give two possible configurations, one Y-shaped and the other T-shaped. In both cases, the general conformation of the glycans is perfectly compatible with their postulated role as a recognition signal.


[Nucleotide sequence determination of yeast mitochondrial phenylalanine-tRNA].

The primary structure of mitochondrial tRNAPhe from Saccharomyces cerevisiae, purified by two-dimensional polyacrylamide gel electrophoresis, was determined using, standard procedures on in vivo 32P-labeled tRNA, as well as the new 5'-end postlabeling techniques. We propose a cloverleaf model which allows for tertiary interaction between cytosine in position 46 and guanine in position 15 and maximizes base pairing in the psi C stem, thus excluding the uracile in position 50 from base pairing in the psi C stem. Comparison of the primary structure of this tRNA with all other known procaryotic, chloroplastic or cytoplasmic tRNAsPhe sequences does not lead to any conclusion about the endosymbiotic theory of mitochondria evolution.


Similarities of ergonovine-induced and spontaneous attacks of variant angina.

Ergonovine has been shown to provoke attacks of variant angina, but a question remains whether spontaneous and ergonovine-induced attacks of variant angina are similar. Seven patients with variant angina undergoing cardiac catheterization were studied during transient episodes of spontaneous and ergonovine-induced rest angina with ST-segment elevation. Clinical, electrocardiographic, left ventricular hemodynamic and coronary angiographic observations were made before and repeated after ergonovine (0.05-0.2 mg I.V.). The character and duration of chest pain were similar during both spontaneous and ergonovine-induced episodes. ST-segment elevation (greater than 1 mm) was present inferiorly in three patients, anteriorly in three patients, and both inferiorly and anteriorly in one patient during both episodes. Mean heart rate and systolic arterial pressure changed little, while left ventricular end-diastolic pressure increased significantly during spontaneous or ergonovine-induced attacks. We observed subtotal or total dynamic obstruction in the left anterior descending (three patients), right coronary arteries (three patients) and both arteries in one patient during both attacks. Thus, in selected patients ergonovine-induced attacks of variant angina were remarkably similar to spontaneous episodes.


The T4/TBG ratio and the investigation of thyroid function.

The relationship between serum thyroxine (T4) and thyroxine-binding globulin (TBG) has been studied in "euthyroid" subjects. The T4/TBG ratios in these subjects has been calculated and compared with those found in patients suffering from myxoedema and thyrotoxicosis. The ratio allows a more precise selection of the borderline cases requiring Thyroid Stimulating Hormone and triiodothyronine assays, particularly when the serum TBG is raised or lowered.