Enantioselective metabolism of primaquine by human CYP2D6

Given the threat of resistance of human malaria parasites, including to artemisinin derivatives, new agents are needed. Chloroquine (CQ) has been the most widely used anti-malarial, and new analogs (CQAns) presenting alkynes and side chain variations with high antiplasmodial activity were evaluated. Six diaminealkyne and diaminedialkyne CQAns were evaluated against CQ-resistant (CQ-R) (W2) and CQ-sensitive (CQ-S) (3D7) Plasmodium falciparum parasites in culture. Drug cytotoxicity to a human hepatoma cell line (HepG2) evaluated, allowed to calculate the drug selectivity index (SI), a ratio of drug toxicity to activity in vitro. The CQAns were re-evaluated against CQ-resistant and -sensitive P. berghei parasites in mice using the suppressive test. Docking studies with the CQAns and the human (Hss LDH) or plasmodial lactate dehydrogenase (Pf LDH) enzymes, and, a β-haematin formation assay were performed using a lipid as a catalyst to promote crystallization in vitro. All tested CQAns were highly active against CQ-R P. falciparum parasites, exhibiting half-maximal inhibitory concentration (IC50) values below 1 μΜ. CQAn33 and CQAn37 had the highest SIs. Docking studies revealed the best conformation of CQAn33 inside the binding pocket of Pf LDH; specificity between the residues involved in H-bonds of the Pf LDH with CQAn37. CQAn33 and CQAn37 were also shown to be weak inhibitors of Pf LDH. CQAn33 and CQAn37 inhibited β-haematin formation with either a similar or a 2-fold higher IC50 value, respectively, compared with CQ. CQAn37 was active in mice with P. berghei, reducing parasitaemia by 100%. CQAn33, -39 and -45 also inhibited CQ-resistant P. berghei parasites in mice, whereas high doses of CQ were inactive. The presence of an alkyne group and the size of the side chain affected anti-P. falciparum activity in vitro. Docking studies suggested a mechanism of action other than Pf LDH inhibition. The β-haematin assay suggested the presence of an additional mechanism of action of CQAn33 and CQAn37. Tests with CQAn34, CQAn37, CQAn39 and CQAn45 confirmed previous results against P. berghei malaria in mice, and CQAn33, 39 and 45 were active against CQ-resistant parasites, but CQAn28 and CQAn34 were not. The result likely reflects structure-activity relationships related to the resistant phenotype.     

Agrobacterium-mediated transformation and plant regeneration of Brassica oleracea var. capitata

An efficient protocol for Agrobacterium tumefaciens-mediated transformation of four commercial cultivars of Brassica oleracea var. capitata is described. A strain of A. tumefaciens LBA4404 with the neomycin phosphotransferase gene (nptII) and a CaMV 35S-peroxidase gene cassette were used for co-cultivation. Preliminary selection of regenerated transgenic plants was performed on kanamycin-containing medium. The frequency of transgenic plants was calculated on the basis of GUS (β-glucuronidase) activity detected by the histochemical X-gluc test. Tissue-specific GUS expression driven by the peroxidase gene promoter in transgenic plants was analysed by GUS staining. The transformation rates of the commercial cultivars of B. oleracea was higher than in previous reports. Southern blot analysis revealed that integration of marker genes occurred in single and multiple loci in the genome. All transgenic plants grew normally after a brief vernalization period and showed stable inheritance of the marker gene. The present study demonstrates that morphologically normal, fertile transgenic plants of B. oleracea can be obtained. Received: 24 August 1999 / Revision received: 23 November 1999 / Accepted: 3 December     

Leaf hydrocarbons of Phacelia species (hydrophyllaceae)

Whole leaf hydrocarbons of six species of Phacelia were investigated for their variability among natural Californian populations, and for adaptive     

Rho kinase is required for CCR7-mediated polarization and chemotaxis of T lymphocytes

A chemokine receptor, CXCR4, and its endogenous ligand, stromal cell-derived factor-1 (SDF-1), have been recognized to be involved in the metastasis of several types of cancers. T140 analogs are peptidic CXCR4 antagonists composed of 14 amino acid residues that were previously developed as anti-HIV agents having inhibitory activity against HIV-entry through its co-receptor, CXCR4. Herein, we report that these compounds effectively inhibited SDF-1-induced migration of human breast cancer cells (MDA-MB-231), human leukemia T cells (Sup-T1) and human umbilical vein endothelial cells at concentrations of 10–100 nM in vitro. Furthermore, slow release administration by subcutaneous injection using an Alzet osmotic pump of a potent and bio-stable T140 analog, 4F-benzoyl-TN14003, gave a partial, but statistically significant (P≤0.05 (t-test)) reduction in pulmonary metastasis of MDA-MB-231 in SCID mice, even though no attempt was made to inhibit other important targets such as CCR7. These results suggest that T140 analogs have potential use for cancer therapy, and that small molecular CXCR4 antagonists could potentially replace neutralizing antibodies as anti-metastatic agents for breast cancer.     

Identifying factors that influence bird richness and abundance on farms

Capsule: Farmers can influence species richness and abundance of typical farmland birds positively, even on rather small farms (20–50?ha) within intensively farmed areas.

Aims: To assess the impact of farm settings, farm characteristics and heterogeneity of habitats on bird species richness and abundance, and to indicate which actions and measures farmers can take to promote farmland birds at a farm level.

Methods: Farmland bird species richness and abundance were modelled as a function of farm settings, farm characteristics and semi-natural habitats on 133 farms. The data were analysed at the farm scale, as this is the ‘operating range’ of a farmer, but also at the territory scale, which represents the range birds (mainly passerines) use during the breeding season. Additionally, effects of the farm variables on species abundance/occurrence were investigated for nine widespread species.

Results: Farmland bird species abundance (but not richness) was elevated on organic compared to non-organic farms. Farmland bird species richness and abundance increased with decreasing mean field size. Crop diversity had positive effects on five species at the territory scale. Several semi-natural habitats, especially hedgerows, were associated with higher bird species richness and abundance at both farm and territory scales. Settlement revealed rather negative effects at the farm scale, but several positive relations at the territory scale.

Conclusion: Birds, especially passerines, are restricted to a small area during the breeding season, and so even small farms can contribute to their protection by growing diverse crops, reducing field size and managing a diversity of semi-natural, uncropped habitats. These measures should ideally be accessible within the relatively small scale of a bird territory.     

Vitalfärbestudien mit Methylrot

Ohne Zusammenfassung     

Health facility characteristics and their relationship to coverage of PMTCT of HIV services across four African countries: the PEARL study

Background

Health facility characteristics associated with effective prevention of mother-to-child transmission of HIV (PMTCT) coverage in sub-Saharan are poorly understood.

Methodology/Principal Findings

We conducted surveys in health facilities with active PMTCT services in Cameroon, Cote d''Ivoire, South Africa, and Zambia. Data was compiled via direct observation and exit interviews. We constructed composite scores to describe provision of PMTCT services across seven topical areas: antenatal quality, PMTCT quality, supplies available, patient satisfaction, patient understanding of medication, and infrastructure quality. Pearson correlations and Generalized Estimating Equations (GEE) to account for clustering of facilities within countries were used to evaluate the relationship between the composite scores, total time of visit and select individual variables with PMTCT coverage among women delivering.Between July 2008 and May 2009, we collected data from 32 facilities; 78% were managed by the government health system. An opt-out approach for HIV testing was used in 100% of facilities in Zambia, 63% in Cameroon, and none in Côte d''Ivoire or South Africa. Using Pearson correlations, PMTCT coverage (median of 55%, (IQR: 33–68) was correlated with PMTCT quality score (rho = 0.51; p = 0.003); infrastructure quality score (rho = 0.43; p = 0.017); time spent at clinic (rho = 0.47; p = 0.013); patient understanding of medications score (rho = 0.51; p = 0.006); and patient satisfaction quality score (rho = 0.38; p = 0.031). PMTCT coverage was marginally correlated with the antenatal quality score (rho = 0.304; p = 0.091). Using GEE adjustment for clustering, the, antenatal quality score became more strongly associated with PMTCT coverage (p<0.001) and the PMTCT quality score and patient understanding of medications remained marginally significant.

Conclusions/Results

We observed a positive relationship between an antenatal quality score and PMTCT coverage but did not identify a consistent set of variables that predicted PMTCT coverage.     

The cobblers stick to their lasts: pollinators prefer native over alien plant species in a multi-species experiment

The majority of plant species rely, at least partly, on animals for pollination. Our knowledge on whether pollinator visitation differs between native and alien plant species, and between invasive and non-invasive alien species is still limited. Additionally, because numerous invasive plant species are escapees from horticulture, the transition from human-assisted occurrence in urbanized habitats to unassisted persistence and spread in (semi-)natural habitats requires study. To address whether pollinator visitation differs between native, invasive alien and non-invasive alien species, we did pollinator observations for a total of 17 plant species representing five plant families. To test whether pollinator visitation to the three groups of species during the initial stage of invasion depends on habitat type, we did the study in three urbanized habitats and three semi-natural grasslands, using single potted plants. Native plants had more but smaller flower units than alien plants, and invasive alien plants had more but smaller flowers than non-invasive alien plants. After accounting for these differences in floral display, pollinator visitation was higher for native than for alien plant species, but did not differ between invasive and non-invasive alien plant species. Pollinator visitation was on average higher in semi-natural than in urbanized habitats, irrespective of origin or status of the plant species. This might suggest that once an alien species has managed to escape from urbanized into more natural habitats, pollinator limitation will not be a major barrier to establishment and invasion.     

Antioxidant,antibacterial, antifungal activities and gas chromatographic fingerprint of fractions from the root bark of Afzelia africana

Background: Afzelia africana is a tropical plant with numerous ethno-medicinal benefits. The plant has been used for the treatment of pain, hernia, fever, malaria, inflammation and microbial infections. Objectives: To perform bioassay-guided fractionation, antioxidant and antimicrobial activities of the bark of Afzelia africana. Methods: Column chromatography fractionation, antioxidant activity (% (2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid (ABTS) and 1,1-diphenyl picrylhydrazyl (DPPH) scavenging activity))), antimicrobial activity (microbroth dilution: Minimum Inhibitory Concentration (MIC), Minimum Bactericidal Concentration (MBC), MBC/MIC ratio), and synergistic activities (Checkerboard assay: Fraction Inhibitory Concentration Index (FICI)). Results: Bioassay-guided fractionation of A. africana produced four fractions that displayed promising free radical scavenging activities in the ABTS (54-93)% and the DPPH (35-76)% assays in the ranking order of F1(93-54)>F4(81-58)>F2(74-58)>F3(72-55) and F3(77-42)>F1(64-46)>F4(55-44)>F2(47-35) respectively at a concentration range of 1.0-0.01 mg/mL. The fraction F1 (MBC: 2.5-5.0 mg/mL) and F4 (MBC: 1.25-10.0 mg/mL) exhibited broad spectrum of superior bactericidal effects than F2 (MBC≥100.0 mg/mL) and F3 (MBC: 12.5-100.0 mg/mL) against Staphylococcus mutans, Staphylococcus aureus, Escherichia coli, fluconazole-resistant Candida albicans, methicillin-resistant S. aureus, Bacillus subtilis, Klebsiella pneumonia, Pseudomonas aeruginosa, Salmonella typhi, and Candida albicans (standard strain). The two most active fractions (F1 and F4) reported synergistic effects (FICI≤0.5) against S. typhi whilst the F4 reported additional synergism against E. coli, K. pneumonia, and S. typhi when combined with ciprofloxacin. Furthermore, the two fractions reported synergistic effects against Escherichia coli, Klebsiella pneumonia, Salmonella typhi, and Pseudomonas aeruginosa when combined with tetracycline whilst F1 reported antifungal synergism against fluconazole resistant Candida albicans when combined with fluconazole and ketoconazole. Conclusion: The study has confirmed the antioxidant, antimicrobial and synergistic uses of A. africana for the treatment of both infectious and non-infectious disease.