Effect of anticancer drugs on the release of interleukin-6 in vitro

Summary This study investigates the effects of anticancer drugs and immunomodulating agents on the release of interleukin-6 (IL-6) from lipopolysaccharide-stimulated human peripheral blood mononuclear leucocytes in vitro. The addition of non-cytotoxic concentrations of Adriamycin (doxorubicin), vincristine and 4-OOH-cyclophosphamide (the in vitro active analogue of cyclophosphamide) resulted in suppression of IL-6 release. The drugs bleomycin, FK156 [d-lactoyl-l-alanyl--d-glutamyl-(l)-meso-diaminopimelyl-(l)-glycine], FK565 [heptanoyl--d-glutamyl-(l)-meso-diaminopimelyl-(d)-alanine] and the immunosuppressive agent cyclosporin A did not alter the release of IL-6 in the same experimental system.     

Drosophila homologs of two mammalian intracellular Ca(2+)-release channels: identification and expression patterns of the inositol 1,4,5-triphosphate and the ryanodine receptor genes.

We have identified and cloned portions of two Drosophila genes homologous to two classes of mammalian intracellular Ca(2+)-release channels, the ryanodine receptor and the inositol 1,4,5-triphosphate (IP3) receptor. The Drosophila ryanodine receptor gene (dry) encodes an approx. 15 kb mRNA. It is expressed in the mesoderm of early stage-9 embryos and subsequently in somatic muscles and their precursor cells. In adults, dry mRNA was detected in tubular muscles and at a lower level in neuronal tissues. Embryonic expression of the Drosophila IP3 receptor gene (dip) appears more dynamic and is associated with developing anterior sense organs. In adults, dip expression occurs in several tissues, and relatively high levels of dip mRNA in adult antennae suggest a role for this gene product during olfactory transduction.     

Studies on effects of tamoxifen (ICI 46474) on agonistic encounters between pairs of intact mice

The anti-estrogen tamoxifen (Tam), which has been shown to dramatically suppress offensive behavior in male rats without markedly influencing other aspects of the social encounter, was tested for its effectiveness in mice. TO strain albino mice were given control injections or 50 or 100 micrograms of Tam for 4 or 8 days. Subsequently, mice were tested in pairs (for a particular dose and treatment duration) in which both animals received Tam, one animal received Tam and one saline, or both animals received saline control injections. Ten-minute videotaped encounters were analyzed in terms of total times allocated to nonsocial investigation, social investigation, offense, defense, sexual activity/intense social investigation, and immobility. The lower dose given for the shorter duration produced less social investigation and more nonsocial investigation when Tam-treated subjects were paired together (cf. the Tam vs saline pairing). At all the other doses and durations, Tam reduced offense. Defense also changed in those pairings, but that activity seemed related to the amount of attack received. Tamoxifen had little influence on the weights of accessory sex glands. The data confirm that Tam is a potent suppressor of "androgen-dependent" aggression in male laboratory mice and provide further support for the aromatization hypothesis.     

A novel multistep method for generating precise unidirectional deletions using BspMI, a class-IIS restriction enzyme

A novel approach is described that permits the introduction of unidirectional deletions into a cloned DNA fragment, in a precisely controlled manner. The method is based on the use of a special vector and a class-IIS restriction endonuclease, BspMI, which produces staggered cuts 4 and 8 nucleotides (nt) to the 3' from its recognition site 5'-ACCTGC-3'. The DNA fragment is inserted into the pUC19-based plasmid, which contains a unique BspMI recognition site, and the appropriate number of cleavage-and-deletion cycles is performed, each cycle removing 4 bp. Since the recognition site is not affected by the BspMI cleavage, no recloning of the DNA fragment is necessary. Each cycle consists of BspMI cleavage, removal of the 4-nt single-stranded cohesive ends with mung bean nuclease (MB), and blunt-end ligation to recircularize the plasmid. The shortened plasmid is reintroduced into the host, after one or after several such 4-bp deletion cycles. When DNA is inserted into the multiple cloning site in the lacZ alpha gene, the progress of 4-bp removal can be followed by determining the Lac phenotype, since removal of multiples of 3 bp retains the reading frame while other kinds of deletions distort (or restore) the reading frame. Loss of pre-existing restriction sites or creation of new ones also permits monitoring the progress of the deletion process.(ABSTRACT TRUNCATED AT 250 WORDS)     

In vitro flowering of Fortunella hindsii (Champ.)

Branch internodes of mature plants and stem internodes of seedlings of Fortunella hindsii flowered in vitro on half-strength MT (Murashige and Tucker 1969) basal medium supplemented with benzyladenine, adenine, 6---dimethylallylaminopurine and kinetin. The highest percentage of flowering was achieved with explants originating from branch internodes of flowering plants close to the apex on half-strength MT basal medium containing 5% sucrose and 0.01 mg 1^–1 BA in light. Exposure to darkness for more than 3 weeks followed by re-exposure to light reduced flowering. Flowering required a 4-day exposure to BA, but shoot formation could be initiated even without exposure to BA. First branch internode segments on MT basal medium containing 5% sucrose were prolific in flower (85%) production. The sucrose treatment affected the flower bud size distribution. There were about 13 flower buds per culture in the largest size category (>5 mm).     

Ultrastructure of rat pituitary LH gonadotrophs in relation to serum and pituitary LH levels following repeated LH-RH stimulation

The effects of single and repeated LH-RH injections at 120 min intervals on female rat LH gonadotrophs and on pituitary and serum LH levels were investigated using electronmicroscopy and radioimmunoassay. A temporary stimulation of granule release, of protein and new granule synthesis and of the accumulation of lysosomal structures was found in LH cells after the first LH-RH injection. The temporary stimulations were massively enhanced after the second injection. These consecutive yet in their time-sequence overlapping processes account for the initial depletion of secretory granule content (3--15 min after LH-RH injection), for the subsequent regranulation and accumulation of granules above control levels (60--120 min after injection) and also for the reduction in the number of granules to control levels (150 min after LH-RH injection and thereafter). Increased polymorphic lysosomal structures are believed to be responsible for this reduction of excess granules. The amount of assayable pituitary and serum LH generally corresponds with the morphological changes observed in LH-gonadotrophs, thus further substantiating the above observations. A schema which summarizes the observed morphological and hormonal changes in their time-sequence in response to LH-RH stimulation depicts the short-term regulation of secretory processes in female gonadotrophs.     

Biology of a buprestid beetleSphenoptera clarescens [Col.: Buprestidae], from skeleton weed,Chondrilla juncea

Investigations carried out in Iran, to discover biological control agents for skeleton weed (Chondrilla juncea L.), showed that the buprestid,Sphenoptera clarescens Kerr, was one of the insects highly damaging to the weed. The larvae of this beetle were active throughout theChondrilla season and fed mainly on the rootstocks, causing a progressive decline in naturalChondrilla populations. The host range of the beetle was examined in the field and in the glasshouse, and it was demonstrated thatS. clarescens occurs on and attacks severalCompositae related toC. juncea. As lettuce,Lactuca sativa L., and endive,Cichorium endivia L. could have been at risk in Australia, studies were discontinued.     

Rhabdospora thelohani Laguessé, 1895 (Apicomplexa): new host and geographic records with taxonomic considerations.

New fish species and geographic records for Rhabdospora thelohani Laguessé, 1895 (rodlet cells) are presented. Additionally, the ultrastructure of R. thelohani in Alburnoides bipunctatus ohridanus Karaman, Borostomias antarcticus (L?nnberg), Leuciscus cephalus albus Bonaparte and Rutilus rubilio (Bonaparte) is compared with that reported by other authors and with members of Subphylum Apicomplexa. The ultrastructure of R. thelohani was similar in all the fish species examined; however, the organism was not present in all members of any single species and had intertissue density variations. Rhabdospora thelohani is pyriform, averaging in size 7 X 12 micrometer, with a basal nucleus. The surface complex is composed of a layer (0.5 micrometer diameter) formed by microfilaments (9.3 nm) and an outer trilaminar membrane (9.3 nm). The cytoplasm contains structures identical to rhoptries, micronemes and subpellicular microtubules. Mitochondria, Golgi apparatus, and rough endoplasmic reticulum were not observed, althouth free ribosomes were present and arranged in a vesicular pattern. The observations suggest that the organism moves between cell of epithelial layers and is either released into a lumen intact or passively or actively discharges its contents into a lumen. Results from this study indicate that R. thelohani should be considered a member of Apicomplexa unless definitive evidence is presented to the contrary.