Acyl carrier protein structural classification and normal mode analysis

All acyl carrier protein primary and tertiary structures were gathered into the ThYme database. They are classified into 16 families by amino acid sequence similarity, with members of the different families having sequences with statistically highly significant differences. These classifications are supported by tertiary structure superposition analysis. Tertiary structures from a number of families are very similar, suggesting that these families may come from a single distant ancestor. Normal vibrational mode analysis was conducted on experimentally determined freestanding structures, showing greater fluctuations at chain termini and loops than in most helices. Their modes overlap more so within families than between different families. The tertiary structures of three acyl carrier protein families that lacked any known structures were predicted as well.     

In vivo rates of erythrocyte glutathione synthesis in adults with sickle cell disease

Despite reports of lower GSH concentration in sickle cell disease (SCD), the in vivo kinetic mechanism(s) responsible for GSH deficiency is unknown. To determine whether suppressed synthesis was responsible for the lower erythrocyte GSH concentration, we used a primed intermittent infusion of [(2)H(2)]glycine to measure erythrocyte GSH synthesis in vivo in 23 individuals with homozygous beta(s) SCD and 8 healthy controls. Erythrocyte cysteine concentration, the rate-limiting precursor for GSH synthesis, plasma markers of oxidant damage, and dietary intakes of energy and protein were also measured. Compared with values of controls, SCD subjects had significantly lower erythrocyte GSH (P < 0.04) and cysteine concentrations (P < 0.004) but significantly faster fractional rates of GSH synthesis (P < 0.02). The absolute rates of GSH synthesis in SCD subjects compared with control subjects was greater by approximately 57% (P = 0.062). However, the concentrations of markers of oxidative damage, plasma derivatives of reactive oxygen metabolites, plasma nitrotyrosine, urinary isoprostane-to-creatinine ratio, and GSH-to-GSSG ratio, as well as dietary intakes of energy, protein, and GSH precursor amino acids, were not different between SCD subjects and controls. The findings of this study suggest that the lower erythrocyte GSH of SCD patients is not due to suppressed synthesis or impaired regeneration but rather to increased consumption. In addition, the lower erythrocyte cysteine concentration plus the faster rate of GSH synthesis strongly suggest that the endogenous cysteine supply is not sufficient to meet all anabolic demands; hence, cysteine may be a conditionally essential amino acid in individuals with SCD.     

Impact of the biological control agent Mesoclanis polana (Tephritidae) on bitou bush (Chrysanthemoides monilifera subsp. rotundata) in eastern Australia

The seed fly Mesoclanis polana (Diptera: Tephritidae) was released in Australia in 1996. Its impact on seed production of bitou bush (Chrysanthemoides monilifera subsp. rotundata) was monitored at eight sites along the New South Wales coast from 1996 to 2004. Peak flowerhead production occurred in autumn (March to May); therefore, samples collected in May of each year were used to compare abundance and impact of M. polana across sites and between years. Latitude had a significant effect on abundance and impact of M. polana. By May 2004, 99.6% of flowerheads at the five most northern sites contained at least one egg, while 64% of flowerheads from the two most southern sites contained at least one egg. In May 2004, mean numbers of M. polana eggs per flowerhead were between 13 and 17 at four of the six northern sites and below two at the two southern sites. In May 2004, average seed destruction by M. polana was 58% at the four most northern sites and 11% at the two most southern sites. The highest level of seed destruction recorded was 86% at Kingscliff in May 2003. Percentage seed destruction in May provided a reasonable estimate of seed destruction for the whole year. Parasitoids were found throughout the range of M. polana in Australia. Their attack rate on M. polana was less than 10% at all sites, except at the two most northern sites where parasitism of up to 27% was recorded. Parasitism results are compared with predictions made following an earlier study. The release of Mesoclanis magnipalpis, a species suited to cooler environments, is recommended for the southern range of C. monilifera subsp. rotundata.     

Large-scale expansion of cytomegalovirus-specific cytotoxic T cells in suspension culture

Clinical trials in recent years involving the adoptive transfer of antigen-specific cytotoxic T lymphocytes (CTL) have shown promise in restoring immunity against viral infection and reducing tumor burden in patients with solid and hematological malignancies. However, the large cell number required to achieve efficacy, 10(9) to 10(11), makes routine application of adoptive immunotherapy impractical. Investigation into new methods of CTL expansion may be useful in addressing this problem. Use of stirred suspension bioreactors are one such method that may allow large-scale T-cell expansion. Suspension cultures offer advantages over conventional static culture methods, including providing a homogeneous culture environment, and the potential for optimization and control of culture conditions. We generated cytomegalovirus (CMV)-specific CTL and investigated the potential of stirred bioreactor systems for expansion of large cell numbers. We found that CTL can be readily expanded ( > 200-fold) from cryopreserved stocks by nonspecific stimulation in the presence of allogeneic feeder cells and interleukin-2 (IL-2). Activated CTL inoculated into either suspension or static cultures could be subsequently expanded tenfold, and showed similar growth kinetics and metabolism independent of the culture vessel used. Furthermore, CTL remained specific for CMVpp65 peptide through the expansion phases, as demonstrated by pp65-tetramer staining ( > 95% tetramer(+)) and cytotoxicity assays. This study indicates that suspension reactor systems may be useful in large-scale expansion of antigen-specific CTL lines or clones, and may facilitate the advancement of routine adoptive immunotherapy.     

Efficacy of ground spray application of bait sprays with malathion or spinosad on Mexican fruit fly (Diptera: Tephritidae) in Texas citrus

An important component in the Mexican fruit fly, Anastrepha ludens (Loew) (Diptera: Tephritidae), eradication program is bait spray application to knock down localized A. ludens infestations enhancing the sterile to wild fly ratio and increasing the effectiveness of the sterile insect release program. Efficacy tests were conducted using spray equipment that applies ultralow application rates of malathion NU-LURE or GF-120 spinosad by ground into citrus. Trapit Dome traps located in fields treated with malathion NU-LURE and GF-120 spinosad high rate caught significantly fewer flies than the control in all replications. Treatments reduced the Mexican fruit fly populations by 99.1 and 92.5% with malathion and 98.2 and 89.9% with GF-120 spinosad high rate. Traps in plots with lower rates of GF-120 reduced fly populations by 76.3 and 74.3% in winter and summer test, respectively. There was no indication of fly repulsion from either malathion or GF-120 spinosad during this test. The bait spray option using ground spray equipment to apply ultra-low rates of either malathion NU-LURE or GF-120 spinosad high rate is a viable cost effective treatment method to treat small acreages for A. ludens. For organic growers, the ground spray equipment is effective in applying GF-120 spinosad at the labeled rates.