Measuring phase shifts in humans following a simulated time-zone transition: agreement between constant routine and purification methods

Twelve healthy participants were studied in an Isolation Unit. For the first 7 (control) days, subjects lived on UK time. Then the clock was advanced by 8 h, mimicking an eastward time-zone transition, and for days 8 to 12, participants lived on this new local time. Two constant routines (participants were not allowed to sleep, were restricted in movement, and ate regular, identical snacks) were undertaken, during the control days (days 3 to 4) and at the end of the experiment (days 11 to 12). Rectal temperature and activity were measured throughout, with activity used to correct the measured temperatures for the direct (masking) effects of the sleep-wake cycle. Phase changes of the temperature rhythm between the constant routines were assessed by cross-correlation and cosinor analysis. During days 8 to 10, the measured temperatures and those that had been corrected (purified) for masking were assessed by the same two methods, and the shifts were extrapolated to predict the values expected during the second constant routine. Individuals differed widely in the phase shifts of the temperature rhythm, but the correlations between the changes measured by constant routines and those estimated by the purification methods were high (r=0.771 to 0.903), and the differences between them were not significantly different from zero (p>0.24). Phase shifts of the measured (masked) temperature rhythm were poorer predictors of the shift obtained from the constant routines (r3+/-4.5 h). Limitations of the methods due to the variability of results are discussed, but we conclude that the mean phase shifts obtained from purified, but not raw, temperature data show acceptable agreement with those found using our version of the constant routine.     

Coenzyme Q and the regulation of intracellular steady-state levels of superoxide in HL-60 cells

The present work was set to study how CoQ concentrations affected steady-state levels of superoxide in a cellular model of partial CoQ(10) deficiency in cultured human myeloid leukemia HL-60 cells. Culturing HL-60 cells in the presence of p-aminobenzoate, a competitive inhibitor of polyprenyl-4-hydroxybenzoate transferase (Coq2p), produced a significant decrease of CoQ(10) levels without affecting cell viability. Concomitant decreases in CoQ-dependent electron transport activity and mitochondrial membrane potential were observed under these conditions. Intracellular superoxide was significantly elevated in cells treated with p-aminobenzoate, both under serum-containing and serum-free conditions, and this effect was reversed by exogenous CoQ(10). A slight increase of superoxide was also observed in CoQ(10)-supplemented cells in the absence of serum. Our results support a requirement for CoQ(10) to control superoxide levels in HL-60 cells. The importance of extramitochondrial sources of superoxide in cells with impaired CoQ(10) biosynthesis is discussed.     

Synthesis of tricyclic analogs of stephaoxocanidine and their evaluation as acetylcholinesterase inhibitors

The synthesis of simplified analogs of the novel isoquinoline alkaloid stephaoxocanidine, carrying the oxazaphenalene ABC-ring system of the natural product, and their activity as inhibitors of the enzyme acetylcholinesterase, are reported. 5,6-Dimethoxy-7H -8-oxa-1-aza-phenalen-9-one (5) was as active as a Narcissus extract enriched in galantamine.     

Genetic variability and phylogeography of the invasive zebra mussel, Dreissena polymorpha (Pallas)

The zebra mussel, Dreissena polymorpha (Pallas), a bivalve species originally native to the Black and Caspian seas, has invaded Ireland in the last decade. Five microsatellite loci were used to investigate genetic diversity and population structure in 10 populations across Europe (Ireland, UK, the Netherlands and Romania) and the Great Lakes (Lake Ontario and Lake St Clair). Levels of allelic diversity and mean expected heterozygosity were high for all populations (mean number of alleles/locus and H(E) were 10-15.2 and 0.79-0.89, respectively). High levels of polymorphism observed in Irish populations suggest that the Irish founder population(s) were large and/or several introductions took place after foundation. Significant deficits of heterozygotes were recorded for all populations, and null alleles were the most probable factor contributing to these deficits. Pairwise comparisons using Fisher exact tests and F(ST) values revealed little genetic differentiation between Irish populations. The UK sample was not significantly differentiated from the Irish samples, most probably reflecting an English origin for Irish zebra mussels. No significant differentiation was detected between the two Great Lakes populations. Our data support a northwest rather than a central or east European source for North American zebra mussels.     

Phylogeographic analysis of the red seaweed Palmaria palmata reveals a Pleistocene marine glacial refugium in the English Channel

Phylogeography has provided a new approach to the analysis of the postglacial history of a wide range of taxa but, to date, little is known about the effect of glacial periods on the marine biota of Europe. We have utilized a combination of nuclear, plastid and mitochondrial genetic markers to study the biogeographic history of the red seaweed Palmaria palmata in the North Atlantic. Analysis of the nuclear rDNA operon (ITS1-5.8S-ITS2), the plastid 16S-trnI-trnA-23S-5S, rbcL-rbcS and rpl12-rps31-rpl9 regions and the mitochondrial cox2-3 spacer has revealed the existence of a previously unidentified marine refugium in the English Channel, along with possible secondary refugia off the southwest coast of Ireland and in northeast North America and/or Iceland. Coalescent and mismatch analyses date the expansion of European populations from approximately 128,000 BP and suggest a continued period of exponential growth since then. Consequently, we postulate that the penultimate (Saale) glacial maximum was the main event in shaping the biogeographic history of European P. palmata populations which persisted throughout the last (Weichselian) glacial maximum (c. 20,000 BP) in the Hurd Deep, an enigmatic trench in the English Channel.     

Identification of the infectious source of an unusual outbreak of histoplasmosis, in a hotel in Acapulco, state of Guerrero, Mexico

Three isolates of Histoplasma capsulatum were identified from mice lung, liver, and spleen inoculated with soil samples of the X hotel's ornamental potted plants that had been fertilized with organic material known as compost. The presence of H. capsulatum in the original compost was detected using the dot-enzyme-linked immunosorbent assay. Nested-PCR, using a specific protein Hcp100 coding gene sequence, confirmed the fungal identification associated with an unusual histoplasmosis outbreak in Acapulco. Although, diversity between the H. capsulatum isolate from the hotel and some clinical isolates from Guerrero (positive controls) was observed using random amplification of polymorphic DNA based-PCR, sequence analyses of H-anti and ole fragment genes revealed a high homology (92-99%) between them.     

Group 8 and 10 metal acetylide naphthalimide dyads

[M]-CC-naphthalimide derivatives {[M] = CpNi(PPh₃), CpFe(dppe), CpRu(dppe) and Cp*Ru(dppe)} have been prepared and their X-ray structures determined. The structures show significant non-linearity of the acetylide link, with concomitant ν_CC at low energy and high intensity. Other properties confirm the strong-donor/strong-naphthalimide acceptor nature of the compounds. All the compounds exhibit an intense MLCT band in the visible spectrum that resolves into two bands in non-polar solvents. Spectroelectrochemistry of the CpFe species shows that the MLCT band disappears upon oxidation, and the appearance of a LMCT band at lower energy.     

An equation to calculate individual muscle contributions to joint stability

The purpose of the current paper was to use the energy approach to develop a simplified equation for quantifying individual muscle contributions to mechanical stability about all three axes of a particular joint. Specific examples are provided for muscles acting about the lumbar spine's L4/L5 joint. The stability equation requires input of: (1) origin and insertion coordinates, relative to the joint of interest, (2) muscle force, and (3) muscle stiffness. The muscle force must be derived from a biomechanical analysis that first results in static equilibrium about all axes being studied. The equation can also accommodate muscles with nodes that change the line of action, with respect to a particular joint, as it passes from the origin to insertion. The results from this equation were compared to those from a Moment approach using more than two million simulated muscles with three-dimensional orientations. The differences between approaches were negligible in all cases. The primary advantage of the current method is that it is very easy to implement into any 2D or 3D biomechanical model of any joint, or system of joints. Furthermore, this approach will be useful in dissecting total joint stability into the individual contributions of each muscle for various systems, joints, postures and recruitment patterns.     

Use of 13C chemical shift surfaces in the study of carbohydrate conformation. Application to cyclomaltooligosaccharides (cyclodextrins) in the solid state and in solution

The anomeric carbon chemical shifts of free cyclomaltohexaose, -heptaose, -octaose, -decaose, and -tetradecaose (alpha-, beta-, gamma-, epsilon-, and eta-cyclodextrin, respectively), and of alpha-cyclodextrin inclusion complexes, both in the solid state and in solution, were computed using ab initio 13C chemical shift surfaces for the D-Glcp-alpha-(1-->4)-D-Glcp linkage as a function of the glycosidic bond dihedral angles. Chemical shift calculations in the solid state used angle pairs measured from cyclodextrin X-ray structures as input. For estimations in the liquid state two different approaches were employed to account for dynamic averaging. In one, the computed solid-state anomeric carbon chemical shifts for each cyclodextrin D-Glcp monomer were simply averaged to obtain an estimate of the 13C shifts in solution. In the other, chemical shifts for the anomeric carbons were determined by averaging back-calculated 13C shift trajectories derived from a series of 5 ns molecular dynamic simulations for the oligosaccharides with explicit representation of water. Good agreement between calculated and experimental 13C shifts was found in all cases. Furthermore, our results show that the ab initio 13C chemical shift surfaces are sufficiently sensitive to reproduce the small variations observed for the anomeric 13C shifts of the different cyclodextrin D-Glcp units in the solid state with excellent accuracy. The use of chemical shift surfaces as tools in conformational studies of oligosaccharides is discussed.