In vitro cytotoxicity of 5-aminosubstituted 20(S)-camptothecins. Part 1.

A number of 5-aminosubstituted 20(S)-camptothecin analogues were prepared via semi-synthesis starting from 20(S)-camptothecin and 9-methoxy 20(S)-camptothecin. In vitro anti-cancer activity of these analogues was determined using 60 human tumor cell line assay. Although water solubility of most of these compounds was improved compared to 20(S)-camptothecin, their anti-cancer activity was considerably diminished. However, only smaller substituents such as methylamine or hydroxylamine as present in 8s and 8t, respectively, showed good activity with improved water solubility.     

Improvement of rifamycin B production using mutant strains of Amycolatopsis mediterranei

Two mutant strains of Amycolatopsis mediterranei VA17 and VA18 were isolated using physical (UV) and chemical (NTG) mutagens gave high rifamycin B than the parent type when grown in the same fermentation medium with a pH of 7.2, temperature 32v°C for a period of 12 days. The cultural conditions of both mutant strains are similar to the parent strain except temperature which was higher by 4v°C. By this mutation and selection study, rifamycin B production was improved from 1400 mg/l to 2450 mg/l.     

Distinct subcellular localisations of the putative inositol 1,3,4,5-tetrakisphosphate receptors GAP1 and GAP1 result from the GAP1 PH domain directing plasma membrane targeting

Inositol 1,3,4,5-tetrakisphosphate (IP₄), is a ubiquitous inositol phosphate that has been suggested to function as a second messenger. Recently, we purified and cloned a putative IP₄ receptor, termed GAP1^IP4BP[1], which is also a member of the GAP1 family of GTPase-activating proteins for the Ras family of GTPases. A homologue of GAP1^IP4BP, called GAP1^m, has been identified [2] and here we describe the cloning of a GAP1^m cDNA from a human circulating-blood cDNA library. We found that a deletion mutant of GAP1^m, in which the putative phospholipid-binding domains (C2A and C2B) have been removed, binds to IP₄ with a similar affinity and specificity to that of the corresponding GAP1^IP4BP mutant. Expression studies of the proteins in either COS-7 or HeLa cells showed that, whereas GAP1^IP4BP is located solely at the plasma membrane, GAP1^m seems to have a distinct perinuclear localisation. By mutational analysis, we have shown that the contrast in subcellular distribution of these two closely related proteins may be a function of their respective pleckstrin homology (PH) domains. This difference in localisation has fundamental significance for our understanding of the second messenger functions of IP₄.     

Chromosome numerical mosaicism in some hybrids of theSolanum nigrum complex

The colchicine amphiploid (2n=72) of the hybridSolanum nodiflorum (2n=24)×S. villosum (2n=48) was crossed toS. memphiticum (2n=72) and toS. nigrum (2n=72). In the resulting F₁ hybrids chromosome numerical mosaicism was observed in the anthers of some plants. It was found that chromosome mosaicism did not affect chromosome pairing in normal cells. Disharmonious gene combination seems to be responsible for premeiotic somatic instability which results in chromosome mosaicism observed during meiosis in pollen mother cells. Gene or genes for fertility are transmitted independent of those for chromosome stability. It is suggested thatS. nigrum andS. memphiticum possess in common a gene complex, which when combined withS. nodiflorum andS. villosum genomes will induce somatic instability.     

Potential regulation of ADP-ribosylation factor 6 signalling by phosphatidylinositol 3,4,5-trisphosphate

In this review we have described data supporting the conclusion that PtdIns(3,4,5)P3 may regulate the activation state of ARF6 through an ability to recruit the ARF exchange factors ARNO, GRP1 and cytohesin-1 to the plasma membrane. The downstream consequences of such a PtdIns(3,4,5)P3-dependent activation of ARF6 are presently unclear. However, given the role of ARF6 in fusion events at the plasma membrane, we have proposed that PtdIns(3,4,5)P3 may regulate vesicle trafficking at this membrane through its ability to activate ARF6. This is an attractive possibility given the number of PtdIns(3,4,5)P3-dependent pathways which involve some aspect of vesicle trafficking at the plasma membrane, for instance glucose transport, membrane ruffling and cell movement.     

Interaction effects of insecticide combinations on the growth ofScenedesmus bijugatus andSynechococcus elongatus

The interaction effects of insecticide combinations involving an organophosphate (monocrotophos or quinalphos) and a pyrethroid (cypermethrin or fenvalerate) were determined on the growth ofScenedesmus bijugatus (unicellular green alga) andSynechococcus elongatus (unicellular cyanobacterium). In general, an organophosphate and a pyrethroid in combinations interacted additively, antagonistically, and synergistically on the growth of the two phototrophic microorganisms. However, combinations of quinalphos and cypermethrin or fenvalerate yielded additive interaction response in onlyS. bijugatus.     

Study on bioactive compounds from Streptomyces sp. ANU 6277

An attempt was made to study the bioactive compounds from a terrestrial Streptomyces sp. ANU 6277 isolated from laterite soil. Four active fractions were recovered from the solvent extracts obtained from the culture broth of five day-old strain. Three bioactive compounds were purified and identified as 3-phenylpropionic acid, anthracene-9,10-quinone and 8-hydroxyquinoline. The components of the partially purified fourth active fraction were analyzed by gas chromatography-mass spectrometry and identified as benzyl alcohol, phenylethyl alcohol and 2H-1, 4-benzoxazin-3 (4H)-one. Four active fractions were screened for antimicrobial activity against Gram-positive and Gram-negative bacteria, and fungi including phytopathogenic, toxigenic and dermatophytic genera. Among these metabolites, 8-hydroxyquinoline exhibited strong antibacterial and antifungal activity as compared to 3-phenylpropionic acid and anthracene-9,10-quinone.     

Three-dimensional solution structure of <Emphasis Type="Italic">Tropidechis carinatus</Emphasis> venom extract trocarin: a structural homologue of Xa and prothrombin activator

Trocarin belongs to group D of prothrombin activators derived from snake venom of Tropidechis carinatus and is a rich non-hepatic source of Xa, the only known hepatic prothrombin activator. The structural and functional similarity with Xa makes trocarin an interesting target for exploring the structure-functional relationship with Xa. Herein we report a predicted complete three-dimensional all-atom structural model of trocarin equilibrated in explicit water using 4 ns of molecular dynamics simulation. The tertiary structure was modeled using the structure of human blood coagulation factor Xa. The conformational and structural features of trocarin are then compared with the X-ray crystal and solution simulation structures of human factor Xa. The modeled structure of trocarin has four individual domains (Gla, EGF1, EGF2 and SP) connected along the long axis with similar secondary structural elements to Xa. The simulations suggest that sodium ion binding in the serine protease domain is impaired in trocarin as compared to Xa. In contrast to Xa, for which the sodium ion forms an octahedral coordination network that brings two loop regions connecting four anti-parallel beta-sheets together, we do not find a similar pattern of network in trocarin. We observe that the difference in the binding pattern of sodium ion leads to a approximately 2-A "shrinkage" of the beta2 strand (B2), in comparison to human Xa, as marked by a shorter distance between 189Asp373 (S1-site residue) and 195Ser379 (active-site residue) in the B2 strand. We propose that these differences may be linked to the experimentally observed lower amidolytic activity of trocarin as compared to Xa.