Determining the evolutionary potential of a gene

In addition to information for current functions, the sequence of a gene includes potential information for the evolution of new functions. The wild-type ebgA (evolved beta-galactosidase) gene of Escherichia coli encodes a virtually inactive beta-galactosidase, but that gene has the potential to evolve sufficient activity to replace the lacZ gene for growth on the beta-galactoside sugars lactose and lactulose. Experimental evidence, which has suggested that the evolutionary potential of Ebg enzyme is limited o two specific amino acid replacements, is limited to examining the consequences of single base- substitutions. Thirteen beta-galactosidases homologous with the Ebg beta-galactosidase are widely dispersed, being found in gram-negative and gram-positive eubacteria and in a eukaryote. A comparison of Ebg beta-galactosidase with those 13 beta-galactosidases shows that Ebg is part of an ancient clade that diverged from the paralogous lacZ beta- galactosidase over 2 billion years ago. Ebg differs from other members of its clade at only 2 of the 15 active-site residues, and the two mutations required for full Ebg beta-galactosidase activity bring Ebg into conformity with the other members of its clade. We conclude that either these are the only acceptable amino acids at those positions, or all of the single-base-substitution replacements that must arise as intermediates on the way to other acceptable amino acids are so deleterious that they constitute a deep selective valley that has not been traversed in over 2 billion years. The evolutionary potential of Ebg is thus limited to those two replacements.      

Poor physical fitness is independently associated with mild cognitive impairment in elderly Koreans

The purpose of this study was to investigate the association between physical fitness and mild cognitive impairment (MCI) in elderly Koreans. This was a cross-sectional study that involved 134 men and 299 women aged 65 to 88 years. Six senior fitness tests were used as independent variables: 30 s chair stand for lower body strength, arm curl for upper body strength, chair-sit-and-reach for lower body flexibility, back scratch for upper body flexibility, 8-ft up-and-go for agility/dynamic balance, and 2-min walk for aerobic endurance. Global cognitive function was assessed using the Korean version of the Mini-Mental State Examination (MMSE). Potential covariates such as age, education levels, blood lipids, and insulin resistance (IR) markers were also assessed. Compared to individuals without MMSE-based MCI, individuals with MMSE-based MCI had poor physical fitness based on the senior fitness test (SFT). There were significant positive trends observed for education level (p=0.001) and MMSE score (p<0.001) across incremental levels of physical fitness in this study population. Individuals with moderate (OR=0.341, p=0.006) and high (OR=0.271, p=0.007) physical fitness based on a composite score of the SFT measures were less likely to have MMSE-based MCI than individuals with low physical fitness (referent, OR=1). The strength of the association between moderate (OR=0.377, p=0.038) or high (OR=0.282, p=0.050) physical fitness and MMSE-based MCI was somewhat attenuated but remained statistically significant even after adjustment for the measured compounding factors. We found that poor physical fitness was independently associated with MMSE-based MCI in elderly Koreans.     

Molecular dynamics of synthetic leucine-serine ion channels in a phospholipid membrane

Molecular dynamics calculations were carried out on models of two synthetic leucine-serine ion channels: a tetrameric bundle with sequence (LSLLLSL)(3)NH(2) and a hexameric bundle with sequence (LSSLLSL)(3)NH(2). Each protein bundle is inserted in a palmitoyloleoylphosphatidylcholine bilayer membrane and solvated by simple point charge water molecules inside the pore and at both mouths. Both systems appear to be stable in the absence of an electric field during the 4 ns of molecular dynamics simulation. The water motion in the narrow pore of the four-helix bundle is highly restricted and may provide suitable conditions for proton transfer via a water wire mechanism. In the wider hexameric pore, the water diffuses much more slowly than in bulk but is still mobile. This, along with the dimensions of the pore, supports the observation that this peptide is selective for monovalent cations. Reasonable agreement of predicted conductances with experimentally determined values lends support to the validity of the simulations.     

Aufreinigung von Fumonisinen und ihren Hydrolysaten über ein neuartiges Festphasenextraktions-Material

Tests with various clean-up materials after optimisation of different parameters showed that the use of Oasis® material resulted in matrixless chromatograms in HPLC-FLD. The selectivity and detection limit of the method was improved by using LC-MS/MS as the detection system. Mean recovery was 100%, and no negative food matrix effects could be observed.     

The genome sequence of the protostome Daphnia pulex encodes respective orthologues of a neurotrophin, a Trk and a p75NTR: Evolution of neurotrophin signaling components and related proteins in the bilateria

Background

Neurotrophins and their Trk and p75NTR receptors play an important role in the nervous system. To date, neurotrophins, Trk and p75NTR have only been found concomitantly in deuterostomes. In protostomes, homologues to either neurotrophin, Trk or p75NTR are reported but their phylogenetic relationship to deuterostome neurotrophin signaling components is unclear. Drosophila has neurotrophin homologues called Spätzles (Spz), some of which were recently renamed neurotrophins, but direct proof that these are deuterostome neurotrophin orthologues is lacking. Trks belong to the receptor tyrosine kinase (RTK) family and among RTKs, Trks and RORs are closest related. Flies lack Trks but have ROR and ROR-related proteins called NRKs playing a neurotrophic role. Mollusks have so far the most similar proteins to Trks (Lymnaea Trk and Aplysia Trkl) but the exact phylogenetic relationship of mollusk Trks to each other and to vertebrate Trks is unknown. p75NTR belongs to the tumor necrosis factor receptor (TNFR) superfamily. The divergence of the TNFR families in vertebrates has been suggested to parallel the emergence of the adaptive immune system. Only one TNFR representative, the Drosophila Wengen, has been found in protostomes. To clarify the evolution of neurotrophin signaling components in bilateria, this work analyzes the genome of the crustacean Daphnia pulex as well as new genetic data from protostomes.

Results

The Daphnia genome encodes a neurotrophin, p75NTR and Trk orthologue together with Trkl, ROR, and NRK-RTKs. Drosophila Spz1, 2, 3, 5, 6 orthologues as well as two new groups of Spz proteins (Spz7 and 8) are also found in the Daphnia genome. Searching genbank and the genomes of Capitella, Helobdella and Lottia reveals neurotrophin signaling components in other protostomes.

Conclusion

It appears that a neurotrophin, Trk and p75NTR existed at the protostome/deuterostome split. In protostomes, a "neurotrophin superfamily" includes Spzs and neurotrophins which respectively form two paralogous families. Trks and Trkl proteins also form closely related paralogous families within the protostomian RTKs, whereby Trkls are absent in deuterostomes. The finding of p75NTR in several protostomes suggests that death domain TNFR superfamily proteins appeared early in evolution.     

Enhancement of vaccinia virus based oncolysis with histone deacetylase inhibitors

Histone deacetylase inhibitors (HDI) dampen cellular innate immune response by decreasing interferon production and have been shown to increase the growth of vesicular stomatitis virus and HSV. As attenuated tumour-selective oncolytic vaccinia viruses (VV) are already undergoing clinical evaluation, the goal of this study is to determine whether HDI can also enhance the potency of these poxviruses in infection-resistant cancer cell lines. Multiple HDIs were tested and Trichostatin A (TSA) was found to potently enhance the spread and replication of a tumour selective vaccinia virus in several infection-resistant cancer cell lines. TSA significantly decreased the number of lung metastases in a syngeneic B16F10LacZ lung metastasis model yet did not increase the replication of vaccinia in normal tissues. The combination of TSA and VV increased survival of mice harbouring human HCT116 colon tumour xenografts as compared to mice treated with either agent alone. We conclude that TSA can selectively and effectively enhance the replication and spread of oncolytic vaccinia virus in cancer cells.     

Voluntary anonymous linked study of the prevalence of HIV infection and hepatitis C among inmates in a Canadian federal penitentiary for women.

OBJECTIVE: To determine the seroprevalence of HIV infection and hepatitis C among inmates of a federal penitentiary for women. DESIGN: Voluntary, anonymous, linked, point-prevalence study involving testing of blood samples for antibodies to HIV and hepatitis C virus. PARTICIPANTS: All inmates of the multilevel security federal Prison for Women, Kingston, Ont., who volunteered to participate in the study. Inmates at this long-stay facility are from across Canada. OUTCOME MEASURE: Seroprevalence rate among participants of antibodies to HIV and hepatitis C virus. RESULTS: Of the 130 inmates available for study 113 (86.9%) agreed to donate a blood sample. One woman (0.9%) was HIV positive; 45 (39.8%) were positive for hepatitis C antibody. CONCLUSIONS: It is possible to obtain a high participation rate in a voluntary, anonymous, linked point-prevalence study in a long-stay penitentiary. The HIV seroprevalence rate of 0.9% is lower than that found in studies in provincial (short-stay) prisons. However, the high rate of antibodies to hepatitis C suggests a significant level of risk behaviour, most likely injection drug use, and suggests the potential for a rapid increase in the rate of HIV infection should the number of newly admitted HIV-positive inmates who use injection drugs rise.     

The effects of shading on kaempferol content and leaf characteristics of five soybean lines

Previous work has shown that the presence of kaempferol triglucoside (K9) in soybean ( Glycine max L. Merr.) leaves is associated with reduced numbers of stomata, especially on the upper surface. In the present test, shade was imposed on soybean plants as a means of altering the level of K9, and thus testing the relationship between quantities of K9 and stomatal density. Five lines of soybean that differ in their complement of flavonol glycosides were grown in the field unshaded and also with a 64 and 80% reduction in daylight intensity. Samples of the second, sixth and tenth trifoliate leaves were taken at 34, 53 and 77 days after planting. Shade reduced the quantities of flavonoids on a per leaf and a per unit area basis. Shade had no effect on stomatal density in lines containing K9, but reduced stomatal density in lines lacking K9. We envisage two opposing effects of shade on stomatal density: a direct effect in which shade reduces numbers, and an indirect effect in which a decrease in K9 content allows an increase in stomatal density. The net effect of shade on lines containing K9 appears to be the maintenance of a constant stomatal density. The quantity of K9 was highly correlated with stomatal density within all treatment combinations, and with all treatments combined.     

Rhythmic emission of floral volatiles from<Emphasis Type="Italic"> Rosa damascena semperflorens</Emphasis> cv. ‘Quatre Saisons’

The control of rhythmic emission of floral volatiles emitted from Rosa damascena semperflorens cv. Quatre Saisons throughout floral development under various light regimes was studied. 2-Phenylethanol was the major volatile emitted in addition to monoterpenols, oxidised monoterpenols, monoterpenes and aromatic compounds. All detected volatiles were emitted rhythmically, with maximum peaks coinciding 8–10 h into a 12-h photoperiod. For some compounds a secondary, nocturnal peak was apparent. The primary and secondary maxima both occurred at approximately 24-h intervals. Rhythms appeared to be regulated endogenously: rhythmic emission continued upon exposure to continuous light or continuous darkness, and a phase shift in emission was induced upon inversion of the photoperiod. Additionally, emission continued after flower excision. A similar profile of free volatiles was stored within the floral tissue, together with glycosidic forms of 2-phenylethanol (>99% -d-glucoside), benzyl alcohol, citronellol and geraniol. Regression analysis indicated a significant decrease in glycosylated 2-phenylethanol through the photoperiod. These results suggest that glycosylated volatiles stored within petals may be a source of rhythmically emitted volatiles.Abbreviations CD Continuous darkness - CL Continuous light - 2-PE 2-Phenylethanol - 2-PEG 2-Phenylethyl -d-glucopyranoside     

Bestimmung von Fumonisinen in komplexen Lebensmittelmatrices mittels Accelerated Solvent Extraktion (ASE)

Accelerated solvent extraction (ASE) is an alternative sample extraction procedure for fumonisins in corn and corn products. ASE gave results comparable to that of a draft CEN method, but required less extraction time. Furthermore, ASE gave significantly higher quantitative values than another method reported for extraction of fumonisins (Trucksess et al., 1995).