Quantitative analysis of anticancer 3-deoxyanthocyanidins in infected sorghum seedlings

3-Deoxyanthocyanidins are structurally related to the anthocyanin pigments, which are popular as health-promoting phytochemicals. Here, it is demonstrated that the 3-deoxyanthocyanidins are more cytotoxic on human cancer cells than the 3-hydroxylated anthocyanidin analogues. At 200 microM concentration, luteolinidin reduced the viability of HL-60 and HepG2 cells by 90 and 50%, respectively. Sorghum is a major source of 3-deoxyanthocyanidins, which are present as seed pigments and as phytoalexins responding to pathogen attack. On the basis of the collision-induced dissociation spectra of luteolinidin and apigeninidin, an LC-MS/MS method, operating in multiple-reaction monitoring mode, was developed for the specific detection and accurate quantification of these compounds in complex mixtures, which may be difficult to analyze using absorbance measurements. The results demonstrated that inoculated sorghum seedlings could be utilized for convenient and large-scale production of 3-deoxyanthocyanidins. A quantity of almost 270 microg/g (fresh weight) of luteolinidin was produced 72 h after fungal inoculation of 1-week-old seedlings.     

Consensus nested PCR amplification and sequencing of diverse reptilian, avian, and mammalian orthoreoviruses

The orthoreoviruses are segmented RNA viruses that infect diverse vertebrate host species. While the most common human orthoreovirus, Mammalian Reovirus, is not typically associated with significant disease, the majority of Orthoreovirus species have been shown to cause significant and often fatal disease in reptiles, birds, and primates. There is significant potential for jumping species. A consensus nested-PCR method was designed for investigation of the RNA-dependent RNA polymerase gene of Orthoreovirus and Aquareovirus. This protocol was used to obtain sequencing template from reoviruses of three different vertebrate classes. Bayesian and maximum likelihood phylogenetic analysis found that all viruses analyzed clustered in the genus Orthoreovirus, that reptile reoviruses formed three distinct clusters, and that an African grey parrot reovirus clustered with Nelson Bay virus from bats. This PCR method may be useful for obtaining templates for initial sequencing of novel orthoreoviruses from diverse vertebrate hosts.     

Host-gut microbiota metabolic interactions

The composition and activity of the gut microbiota codevelop with the host from birth and is subject to a complex interplay that depends on the host genome, nutrition, and life-style. The gut microbiota is involved in the regulation of multiple host metabolic pathways, giving rise to interactive host-microbiota metabolic, signaling, and immune-inflammatory axes that physiologically connect the gut, liver, muscle, and brain. A deeper understanding of these axes is a prerequisite for optimizing therapeutic strategies to manipulate the gut microbiota to combat disease and improve health.     

The stomatopod dactyl club: a formidable damage-tolerant biological hammer

Nature has evolved efficient strategies to synthesize complex mineralized structures that exhibit exceptional damage tolerance. One such example is found in the hypermineralized hammer-like dactyl clubs of the stomatopods, a group of highly aggressive marine crustaceans. The dactyl clubs from one species, Odontodactylus scyllarus, exhibit an impressive set of characteristics adapted for surviving high-velocity impacts on the heavily mineralized prey on which they feed. Consisting of a multiphase composite of oriented crystalline hydroxyapatite and amorphous calcium phosphate and carbonate, in conjunction with a highly expanded helicoidal organization of the fibrillar chitinous organic matrix, these structures display several effective lines of defense against catastrophic failure during repetitive high-energy loading events.     

A mutation affecting curd colour in cauliflower (Brassica oleracea L. var. Botrytis DC)

Summary A single gene mutation in cauliflower resulted in the production of an orange coloured curd when heterozygous, and a stunted orange curd when homozygous. The orange heterozygote has commercial potential.     

FACE facility CO2 concentration control and CO2 use in 1990 and 1991

CO₂ treatment level control and CO₂ use are reported for free-air carbon dioxide enrichment (FACE) facility operations at the University of Arizona's Maricopa Agricultural Center in 1990 and 1991. These are required for evaluation of the validity of biological experiments conducted in four replicates of paired experimental and control plots in a large cotton field and the cost-effectiveness of the plant fumigation facility. Gas concentration was controlled to 550 γmol mol^-1 at the center of each experimental plot, just above the canopy. In both years, season-long (April–September) average CO₂ levels during treatment hours (05:00–19:00 h Mountain Standard Time) were 550 γmol mol⁻¹ measured at treatment plot centers when the facility was operating. Including downtime, the season average was 548 γmol mol⁻¹ in 1991. In 1990, the season averages for the four elevated CO₂ treatments varied from 522 to 544 γmol mol⁻¹, owing to extended periods of downtime after lightning damage. Ambient CO₂ concentration during treatment was 370 γmol mol⁻¹. Instantaneous measurements of CO₂ concentration were within 10% of the target concentration of 550 γmol mol⁻¹ more than 65% of the time when the facility was operating, and 1 min averages were within 10% of the target concentration for 90% of the time. The long-term average of CO₂ concentration measured over the 20 m diameter experimental area of one array at the height of the canopy was in the range 550–580 γmol mol⁻¹ during July 1991, with the higher values near the edges. In 1991, CO₂ demand averaged 1250 kg per array per 14 h treatment day, or 4 kg m⁻² of fumigated plant canopy. The FACE facility provided good temporal and spatial control of CO₂ concentration and was a cost-effective method for large-scale field evaluations of the biological effects of CO₂.     

Community level physiological profile response to plant residue additions in Antarctic soils

Changes in microbial community-level physiological profiles (CLPP) were investigated during decomposition of plant materials in maritime Antarctic soils from Alexander Island and Signy Island. For each soil, a series of microcosms were established and half of them amended with Deschampsia antarctica residues and the other half left unamended (controls). Microcosms were destructively sampled at regular intervals and a soil suspension from each microcosm was used to inoculate BIOLOG EcoPlates. The results showed differences between samples due to the treatment (amendment) but no systematic changes due to incubation period. Microbial response in terms of the overall BIOLOG activity in FB soils was greater than in JC and increased in the former during plant decomposition whilst the overall activity of the JC soils remained relatively unchanged. The differences in the microbial community response to plant amendment between FB and JC may be related to the C:N ratio of these soils and to differences in the way in which the different microbial communities respond to sporadic resource inputs.     

Use of C-labelled plant materials and ergosterol, PLFA and NLFA analyses to investigate organic matter decomposition in Antarctic soil

The relationship between organic matter decomposition and changes in microbial community structure were investigated in Antarctic soils using ¹³C-labelled plant materials. Soils with and without labelled Deschampsia antarctica (a native Antarctic grass) were incubated for 42 days and sampled at 0, 7, 14, 21, 28 and 42 days. Changes in microbial community structure were assessed using phospholipid fatty acid analysis (PLFA) and an analysis of the fatty acids associated with the neutral lipid fraction (NLFA). These studies showed that there were no significant changes in PLFA or NLFA profiles over time suggesting no change in microbial community structure during residue decomposition. There was a marked increase however, in ergosterol levels in these soils indicative of growth of the fungal biomass. Analysis of this ergosterol using gas chromatography-mass spectrometry confirmed the transformation of the plant residue by showing the incorporation of ¹³C-plant C into the ergosterol. This incorporation of ¹³C into the ergosterol increased over the incubation period. Importantly, these changes associated with fungal growth were not evident in the analysis of either the PLFA or NLFA fractions thus questioning the reliability of such approaches for studying changes in microbial communities associated with the decomposition of plant residues.