Toxic activity from liquid culture of Colletotrichum acutatum

Colletotrichum acutatum has become an increasingly important plant pathogen worldwide. With this background, a study was carried out to characterize the toxicity of liquid culture media from different isolates and to identify some properties of the toxic principles. Liquid culture media from all isolates were toxic to rubber leaves and induced the anthracnose symptoms. Toxicity of the culture filtrate was not host specific and toxic substances were thermostable. Acetone soluble fraction of the culture filtrates retained the toxic activity and it was effective even at a concentration of 700 μg dry mycelium mass/ml. This revised version was published online in June 2006 with corrections to the Cover Date.     

A comparative study of Phytophthora meadii isolates from rubber (Hevea brasiliensis) plantations in Sri Lanka

Thirty-three Phytophthora meadii isolates were obtained from different Hevea brasiliensisclones grown in different climatic regions in Sri Lanka. Growth, morphology and pathogenicity of the isolates were compared. Growth and pathogenicity levels varied among the isolates. Some isolates obtained from moderately susceptible rubber clones were highly pathogenic, compared to isolates obtained from resistant clones. Highly pathogenic isolates produced a higher number of sporangia on agar at 27 ± 2°C. It was impossible to group isolates according to the clone or the region from where they were obtained.This revised version was published online in October 2005 with corrections to the Cover Date.     

Oxygen consumption by tumor cells during membrane vesicle shedding

The incubation of mastocytoma P815 cells at low temperature (0 degrees C/1-2 hr), with a subsequent shift to greater than or equal to 20 degrees C results in the formation and shedding of membrane vesicles from the tumor cell surfaces. This process, when occurring at physiologic temperature (37 degrees C), mimics the morphological and membrane permeability changes occurring during T-lymphocyte mediated cytolysis of tumor cells. The latter is an oxygen dependent event, but it is not known whether this requirement is at the effector T cell or at the tumor cell level. The present study investigated the oxygen consumption rates of mastocytoma P815 cells induced to shed membrane vesicles by a temperature shift (0 degrees C/1-2 hrs----greater than or equal to 20 degrees C). Results showed that cells undergoing the membrane vesicle shedding process had significantly higher oxygen requirements than control non-shedding cells. Inhibition of the shedding process with deuterium oxide and hexylene glycol, reduced the oxygen consumption rates of low temperature treated cells to the level of control cells. The oxygen consumption rates of the latter were unaffected by these microtubule stabilizing agents. These data indicate that the oxygen required for immune T-cell mediated lysis of tumor cells may be at the target tumor cell level.     

Coaxial Aerodynamically Assisted Bio‐jets: A Versatile Paradigm for Directly Engineering Living Primary Organisms

In this paper, a coaxial jetting methodology is demonstrated as a first example (non‐electric field driven) completely run by aerodynamic forces which are brought about by the application of a differential pressure for the safe handling of primary living organisms by means of jets as encapsulated droplets. Previously this jetting technique in this configuration has only been investigated for processing combinations of liquid‐liquid and liquid‐gas systems. These developmental studies into aerodynamically assisted jets (AAJ) have unearthed a versatile bio‐jetting approach referred to here as coaxial aerodynamically assisted bio‐jetting (CAABJ). In the current work, this flexible approach is demonstrated to handle two primary cell types for drop‐and‐placing onto several different substrates. Furthermore, the study assesses cellular viability of the post‐treated cells in comparison to controls by way of flow cytometry. These first steps demonstrate the promise this protocol has in exploring the creation of biologically viable structures to form encapsulations of cells which would be useful as a direct tissue engineering to the immuno‐hinding methodology in bio‐repair and therapeutics. Therefore, these investigations place CAABJ into the cell jetting pursuit together with bio‐electrosprays, which will undergo an explosive developmental research.     

Environmental influence on the occurrence of epizootic ulcerative syndrome (EUS) in freshwater fish in the Bellanwila–Attidiya wetlands, Sri Lanka

Water quality and rainfall conditions were analysed from November 1996 to September 1997 in selected sampling sites in the Bellanwila–Attidiya wetlands of Sri Lanka, an area where the epizootic ulcerative syndrome (EUS) has occurred, in order to examine the environmental factors that may be associated with EUS. Quantitative sampling of fish populations was carried out over the study period and the recurrence of EUS was monitored by gross clinical signs and histopathology. The occurrence of EUS from the sampling sites was recorded from January 1997 to March 1997. The outbreak was preceded by heavy rainfall. Diurnal variations in air temperature were relatively high during the outbreak period. Declining dissolved oxygen concentrations in water coincided with initiation of the disease outbreak. There were significantly low levels of dissolved oxygen in the water and significantly low rainfall in the area during the period of EUS in comparison with non-EUS periods. The biochemical oxygen demand of the water was significantly higher during the outbreak period than that in the period prior to the outbreak. Of the fish species sampled, the prevalence of EUS was highest in Trichogaster pectoralis (snake skin gourami). Histopathological studies on ulcerative lesions of fish confirmed that diffuse proliferative mycotic granulomatosis is a consistent feature of EUS. The results indicate that interaction between rainfall, deteriorating water quality and presence of pathogens could provide stressful conditions for fish, thereby inducing EUS lesions in susceptible fish populations.     

Perceptions and Patterns of Human–elephant Conflict in Old and New Settlements in Sri Lanka: Insights for Mitigation and Management

Human–elephant conflict poses a major threat to elephants in many parts of Asia, including Sri Lanka. We studied human–elephant conflict in two areas with contrasting scenarios of landuse and conflict, Kahalle and Yala. Kahalle was developed and settled under the Mahaweli irrigation project and the main agricultural practice was irrigated agriculture, with two annual growing seasons. The area was a mosaic of settlements, agriculture, and small forest patches with ill defined human- and elephant-use areas. Elephants ranged within the habitat mosaic year round, occupying remnant forest patches and raiding adjacent crops at night. In contrast, Yala was dominated by a large protected area complex, and the main agricultural methods were slash-and-burn agriculture and rain-fed paddy cultivation. Human- and elephant-use areas were well defined and segregated. The protected area provided elephants with a refuge and food during the rainy season, when the single annual crop was grown. During the dry season, elephants moved into slash-and-burn areas and utilized leftover crops and pioneer vegetation in fallow fields. The landuse pattern and agricultural practices in Yala facilitated co-existence, whereas that in Kahalle led to year round conflict. We suggest that areas managed according to traditional landuse practices should be part of an elephant conservation strategy, where people and elephants have to share resources.     

Global structural changes in annexin 12. The roles of phospholipid,Ca2+, and pH

Ca2+-dependent membrane interaction has long been recognized as a general property of the annexin (ANX) family of proteins. More recently, it has become clear that ANXs can also undergo Ca2+-independent membrane interactions at mildly acidic pH. Here we use site-directed spin labeling in combination with circular dichroism and biochemical labeling methods to compare the structure and membrane topography of these two different membrane-bound forms of ANX12. Our results reveal strong similarities between the solution structure and the structure of the Ca2+-dependent membrane-bound form at neutral pH. In contrast, all Ca2+-independent membrane interactions tested resulted in large scale conformational changes and membrane insertion. Pairs of spin labels that were in close proximity across the interface of different domains of the protein in both the soluble and Ca2+-dependent membrane form were >25 A apart in the Ca2+-independent membrane-bound form. Despite these major conformational changes, the overall secondary structure content did not appear to be strongly altered and ANX12 remained largely helical. Thus, Ca2+-independent membrane interaction leads to massive refolding but not unfolding. Refolding did not occur at low pH in the absence of membranes but occurred within a few seconds after phospholipid vesicles were added. The phospholipid composition of the vesicles was an important modulator of Ca2+-independent membrane interaction. For example, cardiolipin-containing vesicles induced Ca2+-independent membrane interaction even at near neutral pH, thereby raising the possibility that lipid composition could induce relatively rapid Ca2+-independent membrane interaction in vivo.     

Identifying structural features of fibrillar islet amyloid polypeptide using site-directed spin labeling

Pancreatic amyloid deposits, composed primarily of the 37-residue islet amyloid polypeptide (IAPP), are a characteristic feature found in more than 90% of patients with type II diabetes. Although IAPP amyloid deposits are associated with areas of pancreatic islet beta-cell dysfunction and depletion and are thought to play a role in disease, their structure is unknown. We used electron paramagnetic resonance spectroscopy to analyze eight spin-labeled derivatives of IAPP in an effort to determine structural features of the peptide. In solution, all eight derivatives gave rise to electron paramagnetic resonance spectra with sharp lines indicative of rapid motion on the sub-nanosecond time scale. These spectra are consistent with a rapidly tumbling and highly dynamic peptide. In contrast, spectra for the fibrillar form exhibit reduced mobility and the presence of strong intermolecular spin-spin interactions. The latter implies that the peptide subunits are ordered and that the same residues from neighboring peptides are in close proximity to one another. Our data are consistent with a parallel arrangement of IAPP peptides within the amyloid fibril. Analysis of spin label mobility indicates a high degree of order throughout the peptide, although the N-terminal region is slightly less ordered. Possible similarities with respect to the domain organization and parallelism of Alzheimer's amyloid beta peptide fibrils are discussed.