Antibacterial activities of the methanol extracts of Canarium schweinfurthii and four other Cameroonian dietary plants against multi-drug resistant Gram-negative bacteria

Bacterial infections are among the major cause of morbidity and mortality worldwide. The present study was designed to evaluate the in vitro antibacterial activities of the methanol extracts of five Cameroonian edible plants namely Colocasia esculenta, Triumfetta pentandra, Hibiscus esculentus, Canarium schweinfurthii and Annona muricata against a panel of 19 multidrug resistant Gram-negative bacterial strains. The liquid broth microdilution was used to determine the minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) of the extracts. The preliminary phytochemical screening of the extracts was conducted according to the standard phytochemical methods. Results showed that all extracts contained compounds belonging to the classes of polyphenols, triterpenes and steroids, other classes of chemicals being selectively distributed. Canarium schweinfurthii extract showed the best activity with MIC values ranging from 64 to 1024 μg/mL against 89.5% of the 19 tested bacteria strains. MIC values below or equal to 1024 μg/mL were also recorded with Triumfetta pentandra, Annona muricata, Colocasia esculenta and Hibiscus esculentus extracts respectively against 15/19 (78.9%), 11/19 (57.9%), 10/19 (52.6%) and 10/19 (52.6%) tested bacteria. Extract from C. schweinfurthii displayed the lowest MIC value (64 μg/mL) against Escherichia coli AG100A_Tet. Finally, the results of this work provide baseline information for the use of C. esculenta, T. pentandra, H. esculentus, C. schweinfurthii and A. muricata in the treatment of bacterial infections including multidrug resistant phenotypes.     

Direct Measurements of the Pressure and Flow in the Xylem Vessels of Nicotiana tabacum and their Dependence on Flow Resistance and Transpiration Rate

The relationships between xylem tension, velocity of water ascending and transpiration in tobacco plants were measured by means of the “xylem pressure probe technique” (Balling, A. and Zimmermann, U., Planta 182, 325–338, 1990). The flow velocity was determined by suction or injection of fluorescein (or FITC-labelled dextrans of various molecular weights) from the microcapillary of the pressure probe into the punctured xylem vessel, followed by serial-sectioning of the stem after a given propagation time. The distance travelled was defined as the distance from the injection point to the uppermost xylem section in which the dye could be detected. For a transpiration rate of 0.52 ± 0.12 ml . h^?1, a linear dependence between the flow velocity and the tension gradients was found as expected from the Hagen-Poiseuille law. The slope of the straight lines decreased with increasing molecular weight of the fluorescent labelled compound, presumably because of (partial) plugging of the pit membranes. The average value of the flow velocity (2.5 . 10^?4 ± 0.9 . 10^?4 m . s^?1) was one magnitude smaller than the value estimated from the geometric dimensions of the xylem vessels, but agreed well with the literature value of 2.8 . 10^?4 m . s^?1 for herbs (determined by the heat pulse technique; Huber, B. Ber. deutsch. bot. Ges. 50, 89–109, 1932). The average pressure gradient was determined to be 0.39 ± 0.23 MPa . m^?1, in agreement with the literature (Begg, J. E. and Turner, N. C. Plant Physiol. 46, 343–346, 1970). The first response of xylem pressure (or tension) and of flow velocity to a reduction of the transpiration rate (0.14 ± 0.06 ml . h^?1) occurred after about 24 h, when an increase of the xylem pressure towards higher values associated with a decrease in flow velocity was observed. In contrast, re-establishment of the normal transpiration rate brought the pressure (or tension) and the flow velocity back to normal values within half an hour. Similary, introduction of a transverse cross-sectional cut into the stem did not lead during the first 10 h to a change in xylem tension (or velocity). However, during the following day the pressure fell to relatively low values (about ?0.13 MPa). The velocity increased 10-fold. In the next two days the xylem pressure increased again to normal values (average +0.03 MPa), whereas the flow velocity assumed higher values than normal. The data are discussed in terms of the water status and storage of the adjacent tissue cells.     

Antimicrobial peptides: Possible anti-infective agents

Multidrug-resistant bacterial, fungal, viral, and parasitic infections are major health threats. The Infectious Diseases Society of America has expressed concern on the decrease of pharmaceutical companies working on antibiotic research and development. However, small companies, along with academic research institutes, are stepping forward to develop novel therapeutic methods to overcome the present healthcare situation. Among the leading alternatives to current drugs are antimicrobial peptides (AMPs), which are abundantly distributed in nature. AMPs exhibit broad-spectrum activity against a wide variety of bacteria, fungi, viruses, and parasites, and even cancerous cells. They also show potential immunomodulatory properties, and are highly responsive to infectious agents and innate immuno-stimulatory molecules. In recent years, many AMPs have undergone or are undergoing clinical development, and a few are commercially available for topical and other applications. In this review, we outline selected anion and cationic AMPs which are at various stages of development, from preliminary analysis to clinical drug development. Moreover, we also consider current production methods and delivery tools for AMPs, which must be improved for the effective use of these agents.     

Oxidative stress-mediated antimalarial activity of plakortin,a natural endoperoxide from the tropical sponge Plakortis simplex

Plakortin, a polyketide endoperoxide from the sponge Plakortis simplex has antiparasitic activity against P. falciparum. Similar to artemisinin, its activity depends on the peroxide functionality. Plakortin induced stage-, dose- and time-dependent morphologic anomalies, early maturation delay, ROS generation and lipid peroxidation in the parasite. Ring damage by 1 and 10 µM plakortin led to parasite death before schizogony at 20 and 95%, respectively. Treatment of late schizonts with 1, 2, 5 and 10 µM plakortin resulted in decreased reinfection rates by 30, 50, 61 and 65%, respectively. In both rings and trophozoites, plakortin induced a dose- and time-dependent ROS production as well as a significant lipid peroxidation and up to 4-fold increase of the lipoperoxide breakdown product 4-hydroxynonenal (4-HNE). Antioxidants and the free radical scavengers trolox and N-acetylcysteine significantly attenuated the parasite damage. Plakortin generated 4-HNE conjugates with the P. falciparum proteins: heat shock protein Hsp70-1, endoplasmatic reticulum-standing Hsp70-2 (BiP analogue), V-type proton ATPase catalytic subunit A, enolase, the putative vacuolar protein sorting-associated protein 11, and the dynein heavy chain-like protein, whose specific binding sites were identified by mass spectrometry. These proteins are crucially involved in protein trafficking, transmembrane and vesicular transport and parasite survival. We hypothesize that binding of 4-HNE to functionally relevant parasite proteins may explain the observed plakortin-induced morphologic aberrations and parasite death. The identification of 4-HNE-protein conjugates may generate a novel paradigm to explain the mechanism of action of pro-oxidant, peroxide-based antimalarials such as plakortin, artemisinins and synthetic endoperoxides.     

SGT1b is required for HopZ3-mediated suppression of the epiphytic growth of Pseudomonas syringae on N. benthamiana

Type III secreted effectors shape the potential of bacterial pathogens to cause disease on plants. Some effectors affect pathogen growth only in specific niches. For example, HopZ3 causes reduced epiphytic growth of Pseudomonas syringae strain B728a on Nicotiana benthamiana. This raises the question of whether genes important for effector-triggered disease resistance are needed for responses to effectors whose major effect is in the epiphytic niche. We report that SGT1b, a protein known to be important for defense activation, is essential for HopZ3-mediated suppression of PsyB728a epiphytic growth. SGT1b is required for HopZ3- and AvrB3-induced cell death in N. benthamiana plants that express the Pto resistance gene from tomato. We suggest that HopZ3 activates R gene mediated responses in N. benthamiana.