Extrachromosomal human immunodeficiency virus type-1 DNA can initiate a spreading infection of HL-60 cells

In this report, we describe a human immunodeficiency virus type-1 (HIV-1)-infected promyelocytic cell line, OM, derived from HL-60 cells. Although the OM cell line was biologically cloned twice, the pattern of HIV-1 expression during culture appeared analogous to a classical acute spreading infection and was inhibited by both azidothymidine and recombinant soluble CD4 treatment. The number of OM cells actually expressing HIV-1 at the beginning of culture was 0%, reached a peak of nearly 100% at 6 weeks, and then fell to less than 10% HIV-1+ cells by 10 weeks. Clonal analysis of the surviving cells verified that stable HIV-1+ OM cells resulted from the spreading infection. Southern analysis confirmed the transmission of HIV-1 through these OM cultures and the occurrence of stable clones which resulted. The initial percentage of OM cells actually harboring the HIV-1 genome was less than 0.1%, indicating nonfaithful transmission of an unintegrated HIV-1 genome during clonal expansion. These results demonstrate that extrachromosomal HIV-1 DNA can contribute to the spread of HIV-1 infection and give rise to cells which have stably integrated HIV-1 provirus.     

C-methyl phenolics from Qualea species

The trunk wood of Qualea labouriauana contains, besides (2R)-5,7,4′-trihydroxy-3′-methoxy-6,8-dimethylflavanone, (2R)-5,7,4′-trihydroxy-8-methylflavanone, the biosynthetically interesting 2,2′-dihydroxy-4,6,4′,6′-tetramethoxy-3,3′-dimethylbenzophenone. From the trunk wood extract of Q. paraensis the first named flavanone crystallized out directly.     

Hypertrophic cardiomyopathy associated with left ventricular aneurysm and normal coronary arteries: Case study indicating genetic tendencies of cardiomyopathy

A 50-year-old man presented with hypertrophic obstructive cardiomyopathy (HOC) associated with a left ventricular aneurysm and normal coronary arteries. His history revealed no evidence of myocardial infarction or atypical angina. Physical examination disclosed HOC but did not suggest the presence of an aneurysm. Although the patient was treated medically, heart failure ensued, and he died suddenly while working his farm. Subsequent investigation of the patient's family revealed that three of his five children were also affected by cardiomyopathy, which was especially pronounced in the eldest, a 22-year-old man. The possible hemodynamic relationship between HOC and left ventricular aneurysm is discussed, along with probable indications. The role of left ventricular aneurysm is also presented in relation to the natural history of the disease.     

Growth of Thiobacillus novellus on mixed substrates (mixotrophic growth).

In a mixotrophic environment, Thiobacillus novellus concurrently utilized glucose and thiosulfate but showed no stimulation of growth rate or yield. In most mixotrophic environments examined, the growth rate was lower than the heterotrophic growth rate, the extent of the decrease depending on the concentration and relative proportion of thiosulfate and glucose in the medium. Both thiosulfate and glucose were degraded to their most oxidized products in mixotrophic medium, yet the biomass production in this medium was comparable to that found in heterotrophic medium containing glucose alone at the corresponding concentration. It was postulated that in mixotrophic medium the oxidation of thiosulfate, glucose, or partially that of both was uncoupled from energy generation. These results differ in many respects from those reported earlier by LeJohn et al. (J. Bacteriol. 94: 1484--1491, 1967); experiments designed to exactly duplicate some of the growth conditions employed by these workers did not resolve the discrepancy.     

Lytic enzymes in the autolysis of filamentous fungi

The degrees of autolysis attained by five different genera of filamentous fungi during an incubation period of 60 days, under the same culture conditions were: 87.3% for Penicillium oxalicum; 65.9% for Neurospora crassa; 62.7% for Polystictus versicolor; 51.7% for Aspergillus niger and 23.5% for Nectria galligena. N. crassa, A. niger and P. versicolor reached the end of the autolysis during this incubation period (60 days), whereas P. oxalicum and N. galligena did not.The excretion of the lytic enzymes -N-acetyl-glucosaminidase, -1–3 glucanase, chitinase, invertase and acid phosphatase into the culture medium during growth and autolysis was investigated. The excretion of these enzymes was consistent with the degree of autolysis reached, the maximum excretion belonging to P. oxalicum and the minimum to N. galligena. The N. crassa invertase was excreted into the culture liquid at levels very much higher than the other enzymes studied, and at levels very much higher than the invertases excreted by the other fungi.     

Flow cytofluorometric analysis of cell cycle distributions using propidium iodide. Properties of the method and mathematical analysis of the data

In order to better characterize the new rapid staining method for flow cytofluorometry proposed by Krishan, we have tested its stability and several other properties, and have carried out a quantitative comparison of the fluorescence histograms obtained using propidium iodide or the acriflavine-Feulgen staining procedure. Using a human hematopoietic cell line in the logarithmic phase of growth, and analyzing the data by means of a mathematical method we have devised, we found that the fluorescence intentsity of cells stained with propidium iodide remains stable for at least 48 h; it is insensitive to dye concentration between 0.025 and 0.10 mg/ml (37-150 muM); it is not affected by incubation with ribonuclease before staining; propidium iodide in 0.1% sodium citrate remains stable for at least 20 days; and quantitative estimates of the fractions of cells in the different phases of the cell cycle are in good agreement with those obtained from acriflavine-Feulgen staining and from autoradiography after pulse labeling with tritiated thymidine. We conclude that this method is useful for the measurement of relative DNA content by flow cytofluorometry, although modifications in the technique are necessary for some cell types which grow in monolayers.     

Purification and properties of a myotoxin from Conus geographus venom

Previous studies (R. Endean et al. (1974), Toxicon12: 131–138) indicate that whole venom from the marine mollusc Conus geographus directly inhibits skeletal muscle, leaving peripheral nerves, cardiac and smooth muscle unaffected. A quantitative bioassay has been used to detect and measure biologically active myotoxin. By using chromatography on phosphocellulose, purified myotoxin is obtained which has the same physiological effects as whole venom. The LD₅₀ of purified toxin is 12 μg/kg in mice, death occurring as a result of flaccid paralysis and respiratory failure. A biochemical characterization of the purified myotoxin indicates that it is a peptide of 13 amino acids containing two disulfide bonds. This and related peptide myotoxins from Conus should be of exceptional interest in muscle physiology.     

Human urine: Epicatechin metabolites and antioxidant activity after cocoa beverage intake

Associations between cocoa consumption in humans, excreted metabolites and total antioxidant capacity (TAC) have been scarcely investigated. The aims of the study were to investigate the epicatechin (( ? )-Ec) metabolites excreted in urine samples after an intake of 40 g of cocoa powder along with the TAC of these urine samples and the relation between both the analyses. Each of the 21 volunteers received two interventions, one with a polyphenol-rich food (PRF) and one with a polyphenol-free food (PFF) in a randomized cross-over study. Urine samples were taken before and during 24 h at 0–6, 6–12 and 12–24 h periods after test intake. The excreted ( ? )-Ec metabolites and the TAC were determined in urine samples by LC-MS/MS and TEAC assay, respectively. The maximum excretion of ( ? )-Ec metabolites and the maximum TAC value were observed in urine samples excreted between 6 and 12 h after PRF consumption. Significance of TAC increase was found in urine samples excreted during 0–6 and 6–12 h (66.6 and 72.67%, respectively, with respect to the 0 h).