Application of ancient DNA to the reconstruction of past microbial assemblages and for the detection of toxic cyanobacteria in subtropical freshwater ecosystems

Ancient DNA (aDNA) analysis of lake sediments is a promising tool for detecting shifts in past microbial assemblages in response to changing environmental conditions. We examined sediment core samples from subtropical, freshwater Laguna Blanca (Uruguay), which has been severely affected by cultural eutrophication since 1960 and where cyanobacterial blooms, particularly those of the saxitoxin‐producer Cylindrospermopsis raciborskii, have been reported since the 1990s. Samples corresponding to ~1846, 1852, 2000 and 2007 AD were selected to perform denaturing gradient gel electrophoresis (DGGE) analysis of the 16S–23S rRNA intergenic transcribed spacer (ribosomal ITS) to compare their prokaryotic assemblage composition. Each stratum showed different ITS patterns, but the composition of 21st century samples was clearly different than those of mid‐19th century. This compositional change was correlated with shifts in sediment organic matter and chlorophyll a content, which were significantly higher in recent samples. The presence of saxitoxin‐producing cyanobacteria was addressed by quantitative real‐time PCR of the sxtU gene involved in toxin biosynthesis. This gene was present only in recent samples, for which clone libraries and ITS sequencing indicated the presence of Cyanobacteria. Phylogenetic analyses identified C. raciborskii only in the 2000 sample, shortly after several years when blooms were recorded in the lake. These data suggest the utility of aDNA for the reconstruction of microbial assemblage shifts in subtropical lakes, at least on centennial scales. The application of aDNA analysis to genes involved in cyanotoxin synthesis extends the applicability of molecular techniques in palaeolimnological studies to include key microbial community characteristics of great scientific and social interest.     

Alcohol Advertising in Sport and Non-Sport TV in Australia,during Children’s Viewing Times

Estimate the amount of alcohol advertising in sport vs. non-sport programming in Australian free-to-air TV and identify children’s viewing audience composition at different times of the day. Alcohol advertising and TV viewing audience data were purchased for free-to-air sport and non-sport TV in Australia for 2012. We counted alcohol advertisements in sport and non-sport TV in daytime (6am-8.29pm) and evening periods (8.30pm-11.59pm) and estimated viewing audiences for children and young adults (0–4 years, 5–13 years, 14–17 years, 18–29 years). During the daytime, most of the alcohol advertising (87%) was on sport TV. In the evening, most alcohol advertising (86%) was in non-sport TV. There was little difference in the mean number of children (0–17 years) viewing TV in the evening (N = 273,989), compared with the daytime (N = 235,233). In programs containing alcohol advertising, sport TV had a greater mean number of alcohol adverts per hour (mean 1.74, SD = 1.1) than non-sport TV (mean 1.35, SD = .94). Alcohol advertising during the daytime, when large numbers of children are watching TV, is predominantly in free-to-air sport TV. By permitting day-time advertising in sport programs and in any programs from 8.30pm when many children are still watching TV, current regulations are not protecting children from exposure to alcohol advertising.     

Comparative Metal Distribution in Scalp Hair of Pakistani and Irish Referents and Hypertensive Patients

The abnormal metabolism of metal ions plays an important role in health and disease conditions, and studies about them have been attracting significant interest. The aim of our study was to assess the heavy metals (cadmium (Cd), nickel (Ni), lead (Pb), and zinc (Zn)) in scalp hair samples of 50 Irish and 78 Pakistani hypertensive patients of an urban population together with 50 Irish and 96 Pakistani non-hypertensive male subjects in the age group of 30–50 years. The concentrations of trace and toxic elements were measured by inductively coupled plasma–atomic emission spectrophotometer and atomic absorption spectrophotometer before microwave-assisted acid digestion. The validity and accuracy of the methodology were checked using certified reference materials, and by the conventional wet acid digestion method on the same certified reference materials and on real samples. The recovery of all the studied elements was found to be in the range of 97.5–99.7% in certified reference material. The results of this study showed that the mean values of cadmium, nickel, and lead were significantly higher in scalp hair samples of both Pakistani and Irish hypertensive patients than in referents (p < 0.001); whereas, the concentration of zinc was lower in the scalp hair samples of hypertensive patients of both genders. The deficiency of zinc and the high exposure of trace and toxic metals may be the risk factors associated with hypertension.     

cAMP measurements with FRET-based sensors in excitable cells

The development of FRET (fluorescence resonance energy transfer)-based sensors for measuring cAMP has opened the door to sophisticated insights into single-cell cAMP dynamics. cAMP can be measured in distinct cell populations and even in distinct microdomains within cells. However, there is still only limited information on cAMP dynamics in excitable cells, particularly as a function of the activity of voltage-gated Ca2+ channels. A major reason for this is the pH shifts that can occur in excitable cells and their effects on fluorescent proteins.     

Inhibition of platelet adhesion by peptidomimetics mimicking the interactive β-hairpin of glycoprotein Ibα

β-Hairpin peptidomimetics mimicking the interaction sites of the platelet receptor glycoprotein (GP)Ibα with von Willebrand factor (vWF) were synthesised and evaluated for their ability to increase platelet velocity under high shear conditions and to inhibit shear-induced platelet aggregation. A cyclic and bridged dodecapeptide 2e containing a heterochiral diproline motif was identified as a lead compound for the generation of a novel class of potential antiplatelet agents.     

Amino acids in oviduct and uterine fluid and blood plasma during the estrous cycle in the bovine

Up to 40% of cattle embryos die within 3 weeks of fertilization while they are nutritionally dependent on the maternal environment provided by the oviduct and uterine fluids for their development and survival. Despite this dependence there is limited information on the composition of these fluids in cattle. Amino acids are essential for the normal growth and development of the early embryo, acting as precursors of proteins and nucleic acids and as energy sources, osmolytes and signaling molecules. The objective of this study was to measure and compare the amino acid concentrations of oviduct and uterine fluid and blood plasma on different days of the estrous cycle. Oviduct fluid was collected in situ from anaesthetised heifers on Days 0, 2, 3, 4 and 6 and uterine fluid on Days 6, 8 and 14 of the estrous cycle and the concentrations of 19 amino acids determined. Glycine was the most abundant amino acid in both oviduct and uterine fluid. However, the concentrations of many amino acids differed between oviduct and uterus and many were present at higher concentrations in oviduct and uterine fluid than in blood plasma. Oviduct fluid concentrations of amino acids were not affected by day of cycle in contrast to uterine fluid for which there was a day of cycle effect on most of the amino acids. These results provide novel information on the amino acid concentrations in the maternal environment of the early cattle embryo and could form the basis for devising improved media for the production of embryos in vitro.