Infection of human natural killer (NK) cells with replication-defective human T cell leukemia virus type I provirus. Increased proliferative capacity and prolonged survival of functionally competent NK cells.

Human T-cell leukemia virus type I (HTLV-I) can infect a variety of human cell types, but only T lymphocytes are efficiently immortalized after HTLV-I infection. This study reports an attempt to infect and to immortalize NK cells with HTLV-I. Co-cultivation of freshly isolated NK cells with a HTLV-I-producing T cell line did not result in NK cell infection. However, NK cells activated with an anti-CD16 mAb and co-cultivated with a HTLV-I-producing T cell line were reproducibly infected by HTLV-I. HTLV-I infection was documented in NK cell lines and clones by the detection of defective integrated provirus by both Southern blot and polymerase chain reaction analysis. Although HTLV-I-infected NK cells produced viral proteins, they did not produce infectious viral particles. HTLV-I-infected NK cells were phenotypically indistinguishable from their uninfected counterparts (CD16+, CD2+, CD56+, CD3-). They also retained the ability to mediate both natural and antibody-dependent cell cytotoxicity. The IL-2-dependent proliferation of HTLV-I-infected NK cells was significantly greater than that of uninfected NK cells. The doubling time of this infected population was reduced from 9 days to 3 days, and the overall survival of the culture in the absence of restimulation was extended from 5 wk to 18 wk. Unlike T lymphocytes, HTLV-I-infected NK cells were not immortal, implying a fundamental difference between these two lymphocyte populations.     

Absence of VA colonization in Oxalis pes-caprae inoculated with Glomus mosseae

Although members of the Oxalidaceae family have been described as host plants of vesicular-arbuscular mycorrhizal fungi, Oxalis pes-caprae did not become colonized by Glomus mosseae. Extracts of Ox. pes-caprae root inhibited the germination of G. mosseae spores. However, the presence of G. mosseae in the rhizosphere of Ox. pes-caprae produced browning of the roots, which was interpreted as a hypersensitivity response of the plant to the presence of VA fungus.     

Modulation of auxin-binding proteins in cell suspensions

Summary Cultured cell lines from carrot (Daucus carota L.) with little or no embryogenic potential were examined for the auxin-binding capacity of their membranes. The lines belonged to different classes: (a) wild-type lines kept in culture for different periods (the longer the period, the lower being their embryogenic potential); (b) variants, isolated after mutagenesis, showing normal growth but a lack of embryogenic response; (c) auxin-resistant lines, isolated as colonies on solid media containing 45 M 2,4-d; (d) a previously described tumorous line (E9) isolated because of its resistance to hypomethylating drugs. All of these lines showed alterations in auxin-induced, auxin-binding capacity (modulation), i.e. in the non-embryogenic lines the addition of auxin increased the auxinbinding capacity to a very small degree, or removal of the hormone did not produce the proper decrease in that capacity, or both defects could be simultaneously present. Both types of defects were shown to be correctable: after treatments designed to increase the amplitude of modulation, embryogenic capacity was restored in a number of lines.     

Fine structure and histochemistry of the venom gland in the Indian stinging catfish (Heteropneustes fossilis)

The venom glands of Heteropneustes lie deep in the epidermis at the sides of the pectoral fin spines, and consist of large cells that react positively to histochemical tests for proteins, histidine and tyrosine, negatively to periodic acid-Schiff and for serotonin and bombesin. The venom cells are of two types, appearing to have dense or lucent cytoplasm when seen by electron microscopy. The dense type has the better developed Golgi systems, the lucent type has more ribosomes. Both appear to differentiate from the basal layer of the epidermis. The epidermis over the glands has a zone with relatively few desmosomes and enlarged intercellular spaces. Both staining reactions and fine structure differentiate the venom gland cells from the club cells of the epidermis, to such an extent that they must be considered distinct secretory elements.     

Morphological and histochemical study of supragingival human calculus and dental plaque using ruthenium hexammine trichloride and acridine orange.

Ruthenium hexammine trichloride (RHT) and acridine orange were used to preserve and visualize anionic groups in human plaque and dental calculus. RHT-reacting material was present on the membrane of micro-organisms and in intermicrobial spaces of the calcifying areas, and seems to correspond to, and derive from, acidic glyco- and phospholipids of the plasma membrane of the micro-organisms. However, the presence of acidic salivary peptidoglycans cannot be ruled out. Two types of calcification were found: extramicrobial and intramicrobial. The former consisted of calcified deposits irregularly scattered in the intermicrobial matrix. They were in close relationship with RHT-reacting material, or were placed inside vesicular structures delimited by a membrane. Intramicrobial calcification consisted of small aggregates of needle-shaped crystals and/or of granular deposits; in both cases, they either masked the whole cytoplasm of the micro-organisms, or were located only over the plasma membrane. These results suggest that mineral deposition occurs in connection with acidic components of intermicrobial matrix, microbial plasma membranes, and cytoplasms. The addition of RHT and acridine orange to fixing and decalcifying solutions yields satisfactory preservation of dental calculus and plaque, and apparently reduces loss of their anionic organic components and increases their electron density. However, these substances are not sufficient to preserve all ultrastructural details in decalcified areas, probably because the inorganic substance prevents reaction of acridine orange and RHT with the organic components of the calcified matrix.     

Nitrification and denitrification in a system of reciprocating jet bioreactor

Concept of separation of stages coupled with novel design of reciprocating jet bioreactor have been incorporated in this research for the development of high efficiency treatment system for contaminated wastewaters.Evaluation of pilot plant data reveals that a three stage reciprocating jet bioreactor system could be effectively employed for nitrification and denitrification of highly polluted wastewater obtained from Berlin wastewater treatment plant. Such a system with COD destruction stage (residence time 1–3 hours) followed by nitrification stage (residence time 3–4.5 hours) and denitrification stage (residence time 0.3 hours) gives COD destruction rate upto 58 kg COD/(m³ day), nitrification rate upto 3.2 NH ₄ ⁺ -N/(m³ day) and denitrification rate upto 28 kg NO ₃ ^– -N/(m³ day) while providing COD, NH ₄ ⁺ -N and NO ₃ ^– -N conversion of more than 90%.Nitrification and denitrification of wastewater at such a short residence time is possible mainly due to the employment of reciprocating jet bioreactor system.Paper presented at the Third Joint Schlesinger Seminar on Transport phenomena and processes in biological systems, Technion — Israel Institute of Technology, Haifa, Israel, May 8–9, 1990     

Metal exchanges in the fauna-sediment system. The case of Nereis diversicolor and Scrobicularia plana in the Bou Regreg estuary (Morocco)

This study identifies the types and quantities of metals retained by Nereis diversicolor and Scrobicularia plana in a predominantly arid estuary environment.Certain abiotic parameters and a number of physiological processes linked to metabolic and reproductive functions play a decisive role in the seasonal influence of metal levels on the tissues of these organisms.Given the deposivoric diet of the organisms, the sedimentary bed would appear to be the principal source of metal accumulation.The following classification illustrates the bioaccumulation of metals: 209-1The possibility that these metals may be carried into the estuary by sewage water from the cities of Rabat and Sále should certainly not be ignored.     

Mutagenic, electrochemical, and crystallographic investigation of the cytochrome b5 oxidation-reduction equilibrium: involvement of asparagine-57, serine-64, and heme propionate-7

A gene coding for lipase-solubilized bovine liver microsomal cytochrome b5 has been synthesized, expressed in Escherichia coli, and mutated at functionally critical residues. Characterization of the recombinant protein revealed that it has a reduction potential that is approximately 17 mV lower than that of authentic wild-type protein at pH 7 (25 degrees C). Structural studies determined that the recombinant protein differed in sequence from authentic wild-type cytochrome b5 owing to three errors in amidation status in the published sequence for the protein on which the gene synthesis was based. The structural origin of the lower reduction potential exhibited by the triple mutant has been investigated through X-ray crystallographic determination of the three-dimensional structure of this protein and is attributed to the presence of Asp-57 within 3.3 A of heme vinyl-4 in the mutant. In addition, the model developed by Argos and Mathews [Argos, P., & Mathews, F.S. (1975) J. Biol. Chem. 250, 747] for the change in cytochrome b5 oxidation state has been studied through mutation of Ser-64 to Ala. In this model, Ser-64 is postulated to stabilize the oxidized protein through H-bonding interactions with heme propionate-7 that orients this propionate group 6.2 A from the heme iron. Spectroelectrochemical studies of a mutant in which Ser-64 has been changed to an alanyl residue demonstrate that this protein has a reduction potential that is 7 mV lower than that of the wild-type protein; moreover, conversion of the heme propionate groups to the corresponding methyl esters increases the potential by 67 mV.(ABSTRACT TRUNCATED AT 250 WORDS)     

A fluorogenic method for the demonstration of human serum 5′-nucleotide phosphodiesterase isozymes after polyacrylamide gel electrophoresis

A rapid fluorogenic method for the demonstration of 5′-nucleotide phosphodiesterase in human serum has been developed. This method uses the substrate 4-methylumbelliferyl 5′-thymidylate impregnated in agarose gels or filter paper strips. Zymograms are developed in less than 30 min at 25°C, and the sensitivity of this method has been compared with that of the indigogenic method.     

Light-induced pigment granule migration in the retinular cells of Drosophila melanogaster. Comparison of wild type with ERG-defective mutants

The dependence of pigment granule migration (PGM) upon the receptor potential was examined using several strains of electroretinogram (ERG)- defective mutants of Drosophila melanogaster. The mutants that have a defective lamina component but a normal receptor component of the ERG (no on-transient A [nonA] and tan) exhibited normal pigment granule migration. The mutants that have very small or no receptor potentials (certain no receptor potential A [norpA] alleles), on the other hand, exhibited no PGM. In the case of the temperature-sensitive norpA mutant, norpAH52, normal PGM was present at 17 degrees but not at 32 degrees C or above, corresponding to its electrophysiological phenotype. In the transient receptor potential (trp) mutant, whose receptor potential decays to the baseline within a few seconds during a sustained light stimulus, the pigment granules initially moved close to the rhabdomere when light was turned on but moved away after about 5 s during a sustained light stimulus. All these results lend strong support to the notion that PGM is initiated by a light-evoked depolarization of the receptor membrane, i.e., the receptor potential. However, under certain experimental conditions, the receptor potentials failed to induce PGM in the trp mutant. The depolarization of the receptor, thus, appears to be closely associated with PGM but is not a sufficient condition for PGM.