Transmission of freshwater myxozoans during the asexual propagation of invertebrate hosts

The phylum Myxozoa contains over 1350 species almost all of which are considered to be obligate parasites of aquatic animals. The phylum is composed of two classes, the Myxosporea and the Malacosporea, species of which are important pathogens responsible for severe economic losses in cultured fisheries. The life cycles of freshwater Myxozoa are believed to involve horizontal, indirect transmission, involving an invertebrate (oligochaetes or bryozoans) and a vertebrate host (fish or amphibians). Here, we describe myxozoan propagation through the fragmentation of invertebrate hosts to form new infected individuals. The two hosts examined are an oligochaete Lumbriculus variegatus infected with an unidentified myxosporean (Triactinomyxon sp.) and the bryozoan Fredericella sultana infected with the malacosporean Tetracapsuloides bryosalmonae which causes proliferative kidney disease, a major constraint of the European rainbow trout industry. Such intra-clonal propagation is a novel form of vertical transmission that is likely to be widespread within the Myxozoa and could form an important method by which some of these parasites maintain and proliferate within the aquatic environment.     

The high viscosity encountered during freezing in glycerol solutions: effects on cryopreservation

The objective of this study was to determine the viscosity of the residual unfrozen solution that cells are exposed to during freezing in the presence of glycerol and use this to interpret some key aspects of cryopreservation. The viscosity of the glycerol-water binary system exceeded 1000 cP at -40 degrees C, whilst the viscosity of the ternary system, glycerol-water-NaCl, exceeded 100,000 cP at -55 degrees C. The effect of these high viscosities on the diffusion of water at a constant temperature during freezing and during cooling at different linear rates has been estimated. At rates of cooling faster than 100 degrees C min(-1) the diffusion distance during freezing was calculated to be less than 15 microm. Validation of the diffusion calculations was confirmed by examination of the ultrastructure of the freeze concentrated matrix in samples prepared at a range of cooling rates. At a critical rate of cooling, water diffusion becomes limited by the high viscosity and two phenomena, of relevance to cryobiology, occur: (1) the composition of the freeze concentrated matrix around cells deviates from that of the equilibrium phase diagram; and (2) the osmotic loss of water from cells is restricted. These factors are of particular relevance to an understanding of the response of cells such as spermatozoa, red blood cells, and bacteria cooled rapidly with glycerol as cryoprotectant.     

Expression of a pertussis toxin S1 fragment by inducible promoters in oral Streptococcus and the induction of immune responses during oral colonization in mice

Previous work aimed at developing a live oral vaccine expressing pertussis toxin S1 fragment on the surface of the bacterium Streptococcus gordonii elicited a lower than expected antibody response, perhaps because of low antigen expression. In this study, in-frame promoter fusions were constructed to investigate whether an increase in antigen production by the streptococcal vaccine strain results in a better antibody response. The promoters tested were (i) the Streptococcus mutans sucrose-inducible fructosyltransferase (ftf) promoter and (ii) the Bacillus subtilis/Escherichia coli chimeric tetracycline-inducible xyl/tetO promoter. Each of these two promoters was placed upstream of the spaP/s1 fusion gene to drive its expression. The constructs were introduced into S. gordonii DL1 and S. mutans 834. The inducibility of the promoters was confirmed through the determination of SpaP/S1 production via Western blottings. Induced production of SpaP/S1 was observed in S. gordonii and S. mutans with each of the promoters, but the level of expression was the highest in S. mutans, using the xyl/tetO promoter. Thus, S. mutans carrying the xyl/tetO/spaP/s1 construct (S. mutans PM14) was used in oral colonization studies in BALB/c mice. Streptococccus mutans PM14 was able to colonize the animals for the 14-week duration of experimentation. A mucosal IgA response was observed in all the treatment groups but was highest in mice receiving tetracycline induction. In the mouse model of Bordetella pertussis respiratory infection, animals colonized with S. mutans PM14 showed a decreased in B. pertussis lung colony count (P = 0.03) on day 3 compared with control mice colonized by the parent S. mutans 834.     

Traffic of prion protein between different compartments on the neuronal surface, and the propagation of prion disease

The key mechanism in prion disease is the conversion of cellular prion protein into an altered, pathogenic conformation, in which cellular mechanisms play a poorly understood role. Both forms of prion protein are lipid-anchored and reside in rafts that appear to protect the native conformation against conversion. Neurons rapidly traffic their cellular prion protein out of its lipid rafts to be endocytosed via coated pits before recycling back to the cell surface. It is argued in this review that understanding the mechanism of this trafficking holds the key to understanding the cellular role in the conformational conversion of prion protein.     

Structural and functional diversities among mu-conotoxins targeting TTX-resistant sodium channels

mu-Conotoxins are peptides that block sodium channels. Molecular cloning was used to identify four novel mu-conotoxins: CnIIIA, CnIIIB, CIIIA, and MIIIA from Conus consors, C. catus and C. magus. A comparison of their sequences with those of previously characterized mu-conotoxins suggested that the new mu-conotoxins were likely to target tetrodotoxin-resistant (TTX-r) sodium channels. The four peptides were chemically synthesized, and their biological activities were characterized. The new conotoxins all blocked, albeit with varying potencies, TTX-r sodium currents in frog dorsal-root-ganglion (DRG) neurons. The more potent of the four new mu-conotoxins, CnIIIA and CIIIA, exhibited a strikingly different selectivity profile in blocking TTX-r versus TTX-sensitive channels, as determined by their ability to block extracellularly recorded action potentials in three preparations from frog: skeletal muscle, cardiac muscle and TTX-treated C-fibers. CnIIIA was highly specific for TTX-r sodium channels, whereas CIIIA was nonselective. Both peptides appeared significantly less potent in blocking TTX-r sodium currents in rat and mouse DRG neurons. When CnIIIA and CIIIA were injected intracranially into mice, both induced seizures, but only CIIIA caused paralysis. This is the most comprehensive characterization to date of the structural and functional diversities of an emerging group of mu-conotoxins targeting TTX-r sodium channels.     

Morphometric cranial identity of prehistoric Malawians in the light of sub-Saharan African diversity

Little has been described of the Holocene populations of South‐Central Africa, despite the region demonstrating major subsistence shifts relating to dispersals of agriculturalists at least 2,000 years ago. Seven sites with associated human skeletal remains were selected. Hora, Chencherere, Fingura, and Mtuzi represent the Middle Holocene (2,000–5,000 years ago), and Phwadze, Mtemankhokwe, and Nkudzi Bay represent the Late Holocene and the arrival of agriculturalists between 500–2,000 years ago. Focusing on the identity of Hora and Chencherere specimens, two questions were addressed: are the various Holocene Malawians similar to each other, or do they suggest morphological change over time? What modern populations are closest to the prehistoric specimens? The archaeological sample was compared to modern sub‐Saharan Africans from four regions, plus a historic Khoi‐San foraging group. Factor analyses were performed in order to identify complex patterns of variation in metric traits of the skull. According to the results, prehistoric Malawians showed only slight differences between the Late and Middle Holocene, suggesting a population change without any major discontinuity. Later Stone Age skulls did not exclusively show similarities with the Khoi‐San, as they frequently fit well within the variation of modern Bantu‐speaking groups, especially West‐Central Africa. Therefore, we reject the hypothesis that Middle Holocene South‐Central Africans have an exclusively Khoi‐San ancestry, and support an alternative hypothesis that both Middle and Late Holocene groups share a common biological heritage originating in West‐Central Africa in earlier times. Am J Phys Anthropol, 2006. © 2005 Wiley‐Liss, Inc.     

Effects of ribose as an ergogenic aid

The amount of adenosine triphosphate (ATP) stored in the muscle available for immediate use is limited, and once used, must be resynthesized in the muscle. Ribose, a naturally occurring pentose sugar, helps resynthesize ATP for use in muscles. There have been claims that ribose supplements increase ATP levels and improve performance. Other studies have provided mixed results on the effectiveness of ribose as an ergogenic aid at high doses. None of these studies have compared the impact of the recommended dose of ribose on athletes and nonathletes under exercise conditions that are most conducive for effectiveness. The purpose of this study was to evaluate the effectiveness of ribose as an ergogenic aid at the dose recommended for supplements currently on the market during an exercise trial to maximize its efficacy. Male subjects (n = 11) performed 2 trials 1 week apart. Each trial consisted of three 30-second Wingate tests with a 2-minute recovery between each test. Trials were counterbalanced, with 1 trial being performed with 625 mg of ribose and the other with a placebo. Peak power, mean power, and percent decrease in power were recorded during each Wingate test. Repeated-measures analysis of variance (p > 0.05) found no significant differences between ribose and placebo. These results suggest that ribose had no effect on performance when taken orally, at the dose suggested by the distributor.