Catabolism of sulphoquinovosyl diacylglycerol by an enzyme preparation from Phaseolus multiflorus

An enzyme which will deacylate sulphoquinovosyl diacylglycerol (SQDG) has been partially purified from the leaves of runner bean (Phaseolus multiflorus). No monoacyl intermediate was observed and the acyl hydrolase was more active towards unsaturated molecular species of SQDG than towards saturated species. The major peak of activity of SQDG acyl hydrolase, separated on both DEAE-cellulose and Sephadex columns, also contained galactolipid acyl hydrolase activity. The distribution of these activities together with substrate competition and inhibitor experiments indicated that at least part of the SQDG acyl hydrolase activity was due to an enzyme that also hydrolysed galactolipids.     

The effect of trenbolone acetate on the bovine oestrous cycle

The anabolic steroid, trenbolone acetate, affected the bovine oestrous cycle in different ways depending on the timing of administration relative to oestrus and the plasma concentration of trenbolone achieved. Administration before oestrus caused failure of ovulation and a high incidence of a syndrome similar to cystic ovarian disease. High plasma concentrations of trenbolone during the luteal phase of the oestrous cycle prevented luteolysis and maintained progesterone secretion for up to 42 days. Subsequent cycles were not affected.     

The marine cytotoxin portimine is a potent and selective inducer of apoptosis

Portimine is a recently discovered member of a class of marine micro-algal toxins called cyclic imines. In dramatic contrast to related compounds in this toxin class, portimine has very low acute toxicity to mice but is highly cytotoxic to cultured cells. In this study we show that portimine kills human Jurkat T-lymphoma cells and mouse embryonic fibroblasts (MEFs), with LC₅₀ values of 6 and 2.5 nM respectively. Treated cells displayed rapid caspase activation and phosphatidylserine exposure, indicative of apoptotic cell death. Jurkat cells overexpressing the anti-apoptotic protein Bcl-2 or Bax/Bak knockout MEFs were completely protected from portimine. This protection was apparent even at high concentrations of portimine, with no evidence of necrotic cell death, indicating that portimine is a selective chemical inducer of apoptosis. Treatment of the Bcl-2-overexpressing cells with both portimine and the Bcl-2 inhibitor ABT-737 proved a powerful combination, causing >90?% death. We conclude that portimine is one of the most potent naturally derived inducers of apoptosis to be discovered, and it displays strong selectivity for the induction of apoptotic pathways.     

Surface sensing and lateral subcellular localization of WspA,the receptor in a chemosensory‐like system leading to c‐di‐GMP production

Pseudomonas aeruginosa responds to growth on agar surfaces to produce cyclic‐di‐GMP, which stimulates biofilm formation. This is mediated by an alternative cellular function chemotaxis‐like system called Wsp. The receptor protein WspA, is bioinformatically indistinguishable from methyl‐accepting chemotaxis proteins. However, unlike standard chemoreceptors, WspA does not form stable clusters at cell poles. Rather, it forms dynamic clusters at both polar and lateral subcellular locations. To begin to study the mechanism of Wsp signal transduction in response to surfaces, we carried out a structure–function study of WspA and found that its C‐terminus is important for its lateral subcellular localization and function. When this region was replaced with that of a chemoreceptor for amino acids, WspA became polarly localized. In addition, introduction of mutations in the C‐terminal region of WspA that rendered this protein able to form more stable receptor–receptor interactions, also resulted in a WspA protein that was less capable of activating signal transduction. Receptor chimeras with a WspA C‐terminus and N‐terminal periplasmic domains from chemoreceptors that sense amino acids or malate responded to surfaces to produce c‐di‐GMP. Thus, the amino acid sequence of the WspA periplasmic region did not need to be conserved for the Wsp system to respond to surfaces.     

Comparison of Concentration Methods for Quantitative Detection of Sewage-Associated Viral Markers in Environmental Waters

Pathogenic human viruses cause over half of gastroenteritis cases associated with recreational water use worldwide. They are relatively difficult to concentrate from environmental waters due to typically low concentrations and their small size. Although rapid enumeration of viruses by quantitative PCR (qPCR) has the potential to greatly improve water quality analysis and risk assessment, the upstream steps of capturing and recovering viruses from environmental water sources along with removing PCR inhibitors from extracted nucleic acids remain formidable barriers to routine use. Here, we compared the efficiency of virus recovery for three rapid methods of concentrating two microbial source tracking (MST) viral markers human adenoviruses (HAdVs) and polyomaviruses (HPyVs) from one liter tap water and river water samples on HA membranes (90 mm in diameter). Samples were spiked with raw sewage, and viral adsorption to membranes was promoted by acidification (method A) or addition of MgCl₂ (methods B and C). Viral nucleic acid was extracted directly from membranes (method A), or viruses were eluted with NaOH and concentrated by centrifugal ultrafiltration (methods B and C). No inhibition of qPCR was observed for samples processed by method A, but inhibition occurred in river samples processed by B and C. Recovery efficiencies of HAdVs and HPyVs were ∼10-fold greater for method A (31 to 78%) than for methods B and C (2.4 to 12%). Further analysis of membranes from method B revealed that the majority of viruses were not eluted from the membrane, resulting in poor recovery. The modification of the originally published method A to include a larger diameter membrane and a nucleic acid extraction kit that could accommodate the membrane resulted in a rapid virus concentration method with good recovery and lack of inhibitory compounds. The frequently used strategy of viral absorption with added cations (Mg²⁺) and elution with acid were inefficient and more prone to inhibition, and will result in underestimation of the prevalence and concentrations of HAdVs and HPyVs markers in environmental waters.     

Non-crop vegetation associated with chili pepper agroecosystems promote the abundance and survival of aphid predators

Habitat manipulation has long been used as strategy to enhance beneficial insects in agroecosystems. Non-crop weed strips have the potential of supplying food resources to natural enemies, even when pest densities are low. However, in tropical agroecosystems there is a paucity of information pertaining to the resources provided by non-crop weeds and their interactions with natural enemies. In this study we evaluated (a) whether weeds within chili pepper fields affect the diversity and abundance of aphidophagous species; (b) whether there are direct interactions between weeds and aphidophagous arthropods; and (c) the importance of weed floral resources for survival of a native and exotic coccinellid in chili pepper agroecosystems. In the field, aphidophagous arthropods were dominated by Coccinellidae, Syrphidae, Anthocoridae, Neuroptera and Araneae, and these natural enemies were readily observed preying on aphids, feeding on flowers or extrafloral nectaries, and using plant structures for oviposition and/or protection. Survival of native Cycloneda sanguinea (Coleoptera: Coccinellidae) differed between plant species, with significantly greater survival on Ageratum conyzoides and Bidens pilosa. However, no evidence was gathered to suggest that weed floral resources provided any nutritional benefit to the exotic Harmonia axyridis (Coleoptera: Coccinellidae). This research has provided evidence that naturally growing weeds in chili pepper agroecosystems can affect aphid natural enemy abundance and survival, highlighting the need for further research to fully characterize the structure and function of plant resources in these and other tropical agroecosystems.     

A model industrial workhorse: Bacillus subtilis strain 168 and its genome after a quarter of a century

The vast majority of genomic sequences are automatically annotated using various software programs. The accuracy of these annotations depends heavily on the very few manual annotation efforts that combine verified experimental data with genomic sequences from model organisms. Here, we summarize the updated functional annotation of Bacillus subtilis strain 168, a quarter century after its genome sequence was first made public. Since the last such effort 5 years ago, 1168 genetic functions have been updated, allowing the construction of a new metabolic model of this organism of environmental and industrial interest. The emphasis in this review is on new metabolic insights, the role of metals in metabolism and macromolecule biosynthesis, functions involved in biofilm formation, features controlling cell growth, and finally, protein agents that allow class discrimination, thus allowing maintenance management, and accuracy of all cell processes. New ‘genomic objects’ and an extensive updated literature review have been included for the sequence, now available at the International Nucleotide Sequence Database Collaboration (INSDC: AccNum AL009126.4).