Gas exchange responses of a mangrove species, Avicennia marina, to waterlogged and drained conditions

This study was undertaken in summer on fully expanded leaves of Avicennia marina trees in the Beachwood Mangroves Nature Reserve, Durban,South Africa. Data sets were obtained over 5–7 days of relatively dry conditions and over two periods of 5 days during which the swamp was continuously inundated with dilute seawater (< 150 mol m⁻³NaCl). Gas exchange responses were strongly influenced by photosynthetic photon flux density (PPFD), leaf temperature and leaf to air vapour pressure deficit (Δw). Carbon dioxide exchange was saturated at a PPFD of about 800 μmol m⁻² s⁻¹. Maximal CO₂ exchange rates ranged from 8.5 to 9.9 μmol m⁻² s⁻¹ with no differences between drained and waterlogged conditions. Under drained conditions, leaf conductance,transpiration and internal CO₂ concentrations were generally lower, and water use efficiencies higher, than during waterlogging. Continuous waterlogging for 5 days had no adverse effect on CO₂ exchange. Xylem water potentials ranged from −1.32to −3.53 MPa during drained and from −1.02 to −2.65 Mpa during waterlogged conditions. These results are discussed in relation to anatomical and metabolic adaptations of A. marina to waterlogging stress. This revised version was published online in August 2006 with corrections to the Cover Date.     

Coronary artery bypass grafting is associated with immunoparalysis of monocytes and dendritic cells

Coronary artery bypass grafting (CABG) triggers a systemic inflammatory response that may contribute to adverse outcomes. Dendritic cells (DC) and monocytes are immunoregulatory cells potentially affected by CABG, contributing to an altered immune state. This study investigated changes in DC and monocyte responses in CABG patients at 5 time‐points: admission, peri‐operative, ICU, day 3 and day 5. Whole blood from 49 CABG patients was used in an ex vivo whole blood culture model to prospectively assess DC and monocyte responses. Lipopolysaccharide (LPS) was added in parallel to model responses to an infectious complication. Co‐stimulatory and adhesion molecule expression and intracellular mediator production was measured by flow cytometry. CABG modulated monocyte and DC responses. In addition, DC and monocytes were immunoparalysed, evidenced by failure of co‐stimulatory and adhesion molecules (eg HLA‐DR), and intracellular mediators (eg IL‐6) to respond to LPS stimulation. DC and monocyte modulation was associated with prolonged ICU length of stay and post‐operative atrial fibrillation. DC and monocyte cytokine production did not recover by day 5 post‐surgery. This study provides evidence that CABG modulates DC and monocyte responses. Using an ex vivo model to assess immune competency of CABG patients may help identify biomarkers to predict adverse outcomes.     

Morphological and physiological responses of the halophyte, Odyssea paucinervis (Staph) (Poaceae), to salinity

In this study, salt tolerance was investigated in Odyssea paucinervis Staph, an ecologically important C₄ grass that is widely distributed in saline and arid areas of southern Africa. Plants were subjected to 0.2%, 10%, 20%, 40%, 60% and 80% sea water dilutions (or 0.076, 3.8, 7.6, 15.2, 22.8, and 30.4 parts per thousand) for 11 weeks. Increase in salinity from 0.2% to 20% sea water had no effect on total dry biomass accumulation, while further increase in salinity to 80% sea water significantly decreased biomass by over 50%. Morphological changes induced by salinity included reductions in the number of culms, leaves and internodes as well as decreases in internode length and leaf length:leaf width ratios. Carbon dioxide exchange, leaf conductance and transpiration decreased at salinities of 40% and higher, while quantum yield of photosystem II (PSII), electron transport rate (ETR) through PSII and intrinsic photosynthetic efficiency generally decreased at salinities of 60% and higher compared to 0.2% sea water. Concentrations of Na⁺ and Cl⁻ increased significantly with salinity increase in both roots and shoots. Na⁺/K⁺ ratios in the roots and shoots ranged from 0.66 to 3.28 and increased with increase in substrate salinity. The maximal rate of secretion at 80% sea water was 415 nmol cm⁻² d⁻¹ for Na⁺ and 763 nmol m⁻² d⁻¹ for Cl⁻ with high selectivity for these two ions. Predawn and midday ψ decreased with increase in salinity and were more negative than those of the treatment solutions. The concentration of proline increased with increase in salinity in both roots and shoots. The data clearly indicated that O. paucinervis is a highly salt-tolerant species that is morphologically and physiologically adapted to a saline environment.     

Foliar secretory trichomes of Ocimum obovatum (Lamiaceae): micromorphological structure and histochemistry

This study characterises the micromorphology, ultrastructure and main chemical constituents of the foliar glandular trichomes of Ocimum obovatum using light and electron microscopy and a variety of histochemical tests. Two types of glandular trichomes occur on the leaves: large peltate and small capitate. The head of each peltate trichome is made up of four broad head cells in one layer. The head of each capitate trichome is composed of two broad head cells in one layer (type I) or a single oval head cell (type II, rare). In peltate heads, secretory materials are gradually transported to the subcuticular space via fracture in the four sutures at the connecting walls of the head cells. Release to the head periphery occurs through opposite fracture in the four sutures in the head cuticle. In type I capitate trichomes, release of the secretions to the subcuticular space occurs via a pore between the two head cells, and release to the head periphery occurs through the opposite pore in the head cuticle. In type II capitate trichomes, the secreted material is released from the head cell through a ruptured particular squared area at the central part of the head cuticle. These secretion modes are reported for the first time in the family Lamiaceae. Histochemical tests showed that the secretory materials in the glandular trichomes are mainly essential oils, lipophilic substances and polysaccharides. Large peltate trichomes contain a large quantity of these substances than the small capitate trichomes. Ultrastructural evidence suggests that the plastids produce numerous lipid droplets, and the numerous polysaccharide small vesicles are derived from Golgi bodies.     

Reduced glycerol incorporation into phospholipids contributes to impaired intra-erythrocytic growth of glycerol kinase knockout Plasmodium falciparum parasites

Background

Malaria is a devastating disease and Plasmodium falciparum is the most lethal parasite infecting humans. Understanding the biology of this parasite is vital in identifying potential novel drug targets. During every 48-hour intra-erythrocytic asexual replication cycle, a single parasite can produce up to 32 progeny. This extensive proliferation implies that parasites require substantial amounts of lipid precursors for membrane biogenesis. Glycerol kinase is a highly conserved enzyme that functions at the interface of lipid synthesis and carbohydrate metabolism. P. falciparum glycerol kinase catalyzes the ATP-dependent phosphorylation of glycerol to glycerol-3-phosphate, a major phospholipid precursor.

Methods

The P. falciparum glycerol kinase gene was disrupted using double crossover homologous DNA recombination to generate a knockout parasite line. Southern hybridization and mRNA analysis were used to verify gene disruption. Parasite growth rates were monitored by flow cytometry. Radiolabelling studies were used to assess incorporation of glycerol into parasite phospholipids.

Results

Disruption of the P. falciparum glycerol kinase gene produced viable parasites, but their growth was significantly reduced to 56.5 ± 1.8% when compared to wild type parasites. ¹⁴C-glycerol incorporation into the major phospholipids of the parasite membrane, phosphatidylcholine and phosphatidylethanolamine, was 48.4 ± 10.8% and 53.1 ± 5.7% relative to an equivalent number of wild type parasites.

Conclusions

P. falciparum glycerol kinase is required for optimal intra-erythrocytic asexual parasite development. Exogenous glycerol may be used as an alternative carbon source for P. falciparum phospholipid biogenesis, despite the lack of glycerol kinase to generate glycerol-3-phosphate.

General significance

These studies provide new insight into glycerolipid metabolism in P. falciparum.     

Phenotypes and Karyotypes of Human Malignant Mesothelioma Cell Lines

Background

Malignant mesothelioma is an aggressive tumour of serosal surfaces most commonly pleura. Characterised cell lines represent a valuable tool to study the biology of mesothelioma. The aim of this study was to develop and biologically characterise six malignant mesothelioma cell lines to evaluate their potential as models of human malignant mesothelioma.

Methods

Five lines were initiated from pleural biopsies, and one from pleural effusion of patients with histologically proven malignant mesothelioma. Mesothelial origin was assessed by standard morphology, Transmission Electron Microscopy (TEM) and immunocytochemistry. Growth characteristics were assayed using population doubling times. Spectral karyotyping was performed to assess chromosomal abnormalities. Authentication of donor specific derivation was undertaken by DNA fingerprinting using a panel of SNPs.

Results

Most of cell lines exhibited spindle cell shape, with some retaining stellate shapes. At passage 2 to 6 all lines stained positively for calretinin and cytokeratin 19, and demonstrated capacity for anchorage-independent growth. At passage 4 to 16, doubling times ranged from 30–72 hours, and on spectral karyotyping all lines exhibited numerical chromosomal abnormalities ranging from 41 to 113. Monosomy of chromosomes 8, 14, 22 or 17 was observed in three lines. One line displayed four different karyotypes at passage 8, but only one karyotype at passage 42, and another displayed polyploidy at passage 40 which was not present at early passages. At passages 5–17, TEM showed characteristic features of mesothelioma ultrastructure in all lines including microvilli and tight intercellular junctions.

Conclusion

These six cell lines exhibit varying cell morphology, a range of doubling times, and show diverse passage-dependent structural chromosomal changes observed in malignant tumours. However they retain characteristic immunocytochemical protein expression profiles of mesothelioma during maintenance in artificial culture systems. These characteristics support their potential as in vitro model systems for studying cellular, molecular and genetic aspects of mesothelioma.     

Foraging Ranges of Immature African White-Backed Vultures (Gyps africanus) and Their Use of Protected Areas in Southern Africa

Vultures in the Gyps genus are declining globally. Multiple threats related to human activity have caused widespread declines of vulture populations in Africa, especially outside protected areas. Addressing such threats requires the estimation of foraging ranges yet such estimates are lacking, even for widespread (but declining) species such as the African white-backed vulture (Gyps africanus). We tracked six immature African white-backed vultures in South Africa using GPS-GSM units to study their movement patterns, their use of protected areas and the time they spent in the vicinity of supplementary feeding sites. All individuals foraged widely; their combined foraging ranges extended into six countries in southern Africa (mean (± SE) minimum convex polygon area  = 269,103±197,187 km²) and three of the vultures travelled more than 900 km from the capture site. All six vultures spent the majority of their tracking periods outside protected areas. South African protected areas were very rarely visited whereas protected areas in northern Botswana and Zimbabwe were used more frequently. Two of the vultures visited supplementary feeding sites regularly, with consequent reduced ranging behaviour, suggesting that individuals could alter their foraging behaviour in response to such sites. We show that immature African white-backed vultures are capable of travelling throughout southern Africa, yet use protected areas to only a limited extent, making them susceptible to the full range of threats in the region. The standard approach of designating protected areas to conserve species is unlikely to ensure the protection of such wide-ranging species against threats in the wider landscape.     

The secretory apparatus of Xerophyta viscosa (Velloziaceae): Epidermis anatomy and chemical composition of the secretory product

In this study, secretory activity on the adaxial surface of the leaves of the desiccation tolerant plant, Xerophyta viscosa Baker was investigated, using light, scanning and transmission electron microscopy. Glandular activity was associated with sunken cavities which appear to be modifications of infolded epidermal cells. The secretory cavity consisted of a globose lumen surrounded by two layers of cells. The cells of the outer layer were flattened with thickened walls, while those of the inner layer (epithelium), exhibited ultrastructural features of intense metabolic activity. Epithelial cells were larger with a large nucleus, numerous vacuoles, and a dense cytoplasm with abundant mitochondria, endoplasmic reticulum (ER), plastids and a few dictysomes. Lipophilic droplets were abundant in the cytoplasm, mitochondria, ER, plastids and in infolding sites of the plasmalemma outside the protoplast. ER appeared to be involved in the synthesis and transport of lipophilic substances. The mechanism of secretion in X. viscosa appeared to be granulocrine. The chemical composition of hexane and methanol extracts of the leaves, analysed by gas chromatography coupled with mass spectrometry (GC–MS), revealed the presence of diterpenes, phenolic compounds and fatty acids. Compounds in the hexane fraction included velloziolone, manoyl oxide, asperdiol and an unknown substance. Compounds in the methanol fraction included 5-(hydroxymethyl)-2-furancarboxaldehyde, 2,6-dimethoxyphenol, 4-(3-hydroxy-1-propenyl)-2-methoxyphenol and stearic acid.