Antimicrobial activity of extracts of herbal plants used in the traditional medicine of Jordan.

Petroleum ether, ethanol, butanol, and aqueous crude extracts of the whole aerial parts of nine plants exhibited variable degrees of antimicrobial activity against four bacterial and three fungal species. Methanol and hexane extracts did not show any activity. Compared with standard antibiotics, extracts had low to moderate activity. The activity spectrum is wide against gram-positive and negative bacteria as well as fungi tested. However, the butanol extracts at 4 mg/disc of Ononis spinosa (OS), Bryonia syriaca (BS) had high moderate antifungal activity against Aspergillus flavus, Fusarium moniliforme and Candida albicans relative to miconazole nitrate at 40 microg/disc. Furthermore, higher antibacterial activity was observed though low to moderate compared with streptomycin and very comparable with chloramphenicol. Cyclaman persicum (CP) petroleum ether extracts only exhibited pronounced antibacterial activity.

Regulation of the p21-activated kinase-related Dictyostelium myosin I heavy chain kinase by autophosphorylation, acidic phospholipids, and Ca2+-calmodulin.

The Dictyostelium myosin I heavy chain kinase (MIHCK) is a member of the p21-activated kinase family (Lee, S.-F., Egelhoff, T. T., Mahasneh, A., and Côté, G. P. (1996) J. Biol. Chem. 271, 27044-27048). MIHCK incubated with MgATP in the absence of effectors incorporates 1 mol of phosphate/mol, resulting in an approximately 40-fold increase in kinase activity. Sequence analysis of tryptic peptides has identified the major site of phosphorylation as Ser-8. A peptide and a glutathione S-transferase fusion protein containing the Ser-8 phosphorylation site were good substrates for MIHCK, indicating that MIHCK can catalyze its own activation. Guanosine 5'-3-O-(thio)triphosphate (GTPgammaS)-Rac1 stimulates MIHCK autophosphorylation and kinase activity 10-fold. Phosphatidylserine, phosphatidylinositol, and phosphatidylinositol 4,5-bisphosphate, but not phosphatidylcholine or sphingosine, were as effective as GTPgammaS-Rac1 in enhancing MIHCK autophosphorylation and activity. Acidic lipids and GTPgammaS-Rac1 induced the autophosphorylation of a similar set of sites as judged by two-dimensional tryptic peptide maps. It is proposed that GTP-Rac and acidic phospholipids function cooperatively to associate MIHCK with membranes. Ca2+-calmodulin bound MIHCK and inhibited activation by acidic phospholipids but not by GTPgammaS-Rac1. These studies reveal a number of similarities between the regulatory properties of the Dictyostelium and Acanthamoeba MIHCK, suggesting that the signaling pathways that control myosin I are conserved.

Cloning and characterization of a Dictyostelium myosin I heavy chain kinase activated by Cdc42 and Rac.

The motile activities of the small, single-headed class I myosins (myosin I) from the lower eukaryotes Acanthamoeba and Dictyostelium are activated by phosphorylation of a single serine or threonine residue in the head domain of the heavy chain. Recently, we purified a myosin I heavy chain kinase (MIHCK) from Dictyostelium based on its ability to activate the Dictyostelium myosin ID isozyme (Lee, S. -F., and Côté, G. P. (1995) J. Biol. Chem. 270, 11776-11782). The complete sequence of the Dictyostelium MIHCK has now been determined, revealing a protein of 98 kDa that is composed of an amino-terminal domain rich in proline, glutamine, and serine, a putative Cdc42/Rac binding motif, and a carboxyl-terminal kinase catalytic domain. MIHCK shares significant sequence identity with the Saccharomyces cerevisiae Ste20p kinase and the mammalian p21-activated kinase. Gel overlay assays and affinity chromatography experiments showed that MIHCK interacted with GTPgammaS (guanosine 5'-3-O-(thiotriphosphate))-labeled Cdc42 and Rac1 but not RhoA. In the presence of GTPgammaS-Rac1 MIHCK autophosphorylation increased from 1 to 9 mol of phosphate/mol, and the rate of Dictyostelium myosin ID phosphorylation was stimulated 10-fold. MIHCK may therefore provide a direct link between Cdc42/Rac signaling pathways and motile processes driven by myosin I molecules.

Effects of 3,3'-di-O-methylquercetin on guinea-pig isolated smooth muscle.

The effects of the flavone 3,3'-di-O-methylquercetin (DOMQ) have been examined and compared with those of quercetin, on guinea-pig isolated ileum, trachea, and main pulmonary artery (MPA). Except for transient contractions induced by low concentrations (10(-8)-3 x 10(-6) M), DOMQ and quercetin (up to 3 x 10(-4) M) caused reduction of the tone and the phasic contractions of the ileum. A23187 reversed the inhibitory effects of quercetin but not those of DOMQ. DOMQ and quercetin caused concentration-dependent relaxation of the trachea and the adrenaline-contracted MPA. DOMQ shifted to the right the concentration-effect curves induced by acetylcholine on the ileum and the trachea, and by adrenaline on MPA and those induced by CaCl2 on ileum, trachea and MPA. DOMQ also inhibited the contractions induced, in Ca2+-free EGTA-containing buffer, by histamine on ileum and by adrenaline on MPA. These observations suggest that DOMQ inhibits Ca2+ influx, Ca2+ release from intracellular stores and, more likely, Ca2+ binding to intracellular receptor proteins.