Regioselective Enzymatic Hydrolysis of 3',5'-DI-O-Acetylthymidine and Observation of a Surface Effect Due to Polystyrene

The selective enzymatic hydrolysis of 3',5'-di-O-acetylthyidine (1) was studied. The lipases from porcine pancreas and Aspergillus niger, and pig liver esterase, all catalysed selective hydrolysis of the 5'O-acetyl group, but the lipase from Candida cylindracea catalysed selective hydrolysis of the 3'-O-acetyl group. Highest selectivity, leading to essentially pure 3'-O-acetylthymidine, was achieved using porcine pancreatic lipase in dilute solution at pH 7.5. Provision of an artificial interface in the form of polystyrene beads led to a significant increase in the rate of hydrolysis, accompanied by a marked fall in selectivity. Other changes in the hydrolysis conditions, such as raising the concentration of substrate or adding cosolvent, also led to a fall in selectivity.     

Plate tectonics and the problem of anthropoid origins

Using plate tectonic theory, geologic and geophysical determinants of South Atlantic opening are assessed during the Cretaceous through the Oligocene (approximately 135–25 m.y.B.P.). This is done to evalute the feasibility of statements on paleontologic and neontologic distributional data that relate to African and South American separation as the South Atlantic was created. In particular, the question of anthropoid origins is examined in light of plate tectonic theory, and the idea that monophyly of the anthropoid grade is supported by continental drift is refuted. When geological and geophysical evidence is examined in detail, the occurrence of Tertiary transfer of primates between Africa and South America is improbable. Thus one must look to either North America or Middle America for the ancestors of the platyrrhines, and to Eurasia for the ancestors of the catarrhines. The problem of whether those ancestors were prosimians or anthropoids is not dealt with, because the question of whether the Platyrrhini and the Catarrhini are independently evolved from prosimian ancestors (anthropoid diphyly) or are evolved from ancestors that were already of an anthropoid level of structural organization (anthropoid monophyly) can only be resolved on biological grounds and at a level of analysis that allows one to distinguish between analogies developed in parallel, homologies caused by symple-siomorphy, and homologies caused by synapomorphy.     

A protoplast to plant system for the chrysanthemum Dendranthema zawadskii x D. grandiflora

Summary Plantlets were regenerated from protoplasts of in vitro shoot cultures and leaf-derived de novo shoots of the chrysanthemum Dendranthema zawadskii x D. grandiflora. Isolated protoplasts reformed cell walls and then began to divide within 24 hours of culture in streaky plate agarose lenses surrounded by liquid V-KM medium. Twenty one days after isolation, 1 mm diameter callus clumps were transferred to shoot regeneration medium. After a further 33 days leaves became visible. Elongated shoots were rooted on half strength hormone-free MS medium.Abbreviations BAP 6-benzylaminopurine - IAA 3-indoleacetic acid - MS Murashige and Skoog (1962) - NAA 1-naphthylacetic acid - Pfr Photon fluence rate - V-KM Binding and Nehls (1977)     

A sporulation medium forDiscula destructiva,the dogwood anthracnose fungus

A growth medium incorporating finely ground oak or maple leaf tissue was developed for the purpose of increasing in vitro conidium production by the dogwood anthracnose pathogen,Discula destructiva.     

Risks of fetal cystic fibrosis based on linkage disequilibrium data

Summary First-trimester prenatal diagnosis of cystic fibrosis depends on tissues being available from a previous affected child for determination of the phase relationship of DNA markers. If no such tissues are available, it is possible to estimate the risks of a couple producing an affected child from the distribution of haplotypes showing linkage disequilibrium with the cystic fibrosis gene. We have calculated all the fetal risk subsets from the various parental haplotype combinations for the restriction fragment length polymorphisms identified by the KM. 19/PstI and XV-2c/TaqI systems. We conclude that only in a limited number of parental combinations are the fetal risks sufficiently high or sufficiently low to be used in prenatal diagnosis.     

Minimizing Cognitive Errors in Site-Specific Causal Assessments

Interest in causal investigations in aquatic systems has been a natural outgrowth of the increased use of biological monitoring to characterize the condition of resources. Although biological monitoring approaches are critical tools for detecting whether effects are occurring, they do not identify the cause of the observed effects. Formal approaches to causal evaluation can provide a mechanism to build on expert knowledge, increasing the likelihood that remedial efforts will achieve the desired environmental improvement. This paper examines how formal approaches to causal investigations minimize common errors. We reviewed common cognitive errors reported in the literature, and compared them with considerations suggested for strength-of-evidence approaches. Many of the causal considerations are directed toward distinguishing spurious correlations from true causal relationships. However, this is only one type of error; others include hypothesis dependence, confirmation bias, hypothesis tenacity and anchoring. We suggest three general principles for minimizing error in site-specific investigations: (1) Conduct the causal evaluation as a fair, transparent comparison among alternatives; (2) Carefully describe and quantify the conjunction of cause and effect; and (3) Consider that conjunction between cause and effect is spurious, or that a real conjunction was masked.     

High spatial resolution analysis of fungal cell biochemistry – bridging the analytical gap using synchrotron FTIR spectromicroscopy

Fungi impact humans and the environment in many ways, for good and ill. Some fungi support the growth of terrestrial plants or are used in biotechnology, and yet others are established or emerging pathogens. In some cases, the same organism may play different roles depending on the context or the circumstance. A better understanding of the relationship between fungal biochemical composition as related to the fungal growth environment is essential if we are to support or control their activities. Synchrotron FTIR (sFTIR) spectromicroscopy of fungal hyphae is a major new tool for exploring cell composition at a high spatial resolution. Brilliant synchrotron light is essential for this analysis due to the small size of fungal hyphae. sFTIR biochemical characterization of subcellular variation in hyphal composition will allow detailed exploration of fungal responses to experimental treatments and to environmental factors.     

Xenopus exhibits seasonal variation in retinotectal latency but not tecto-isthmo-tectal latency

1. The tectum of Xenopus receives visuotopic input from both eyes. The contralateral eye's projection reaches the tectum directly, via the optic nerve. The ipsilateral eye's projection reaches the tectum indirectly, via the nucleus isthmi and isthmo-tectal projection. 2. Because of the multi-synaptic nature of the ipsilateral pathway, there is an inherent delay between the time that information from the contralateral eye reaches the tectum and the time that information from the ipsilateral eye arrives at the tectum. The length of the intertectal delay is a function of the latencies of the contralateral and ipsilateral pathways. 3. The length of this intertectal delay has functional, as well as developmental, implications with regard to the role of N-methyl-D-aspartate receptors in tectal cell activity and development of orderly synaptic connections. 4. We have found that the latencies of the contralateral and ipsilateral pathways exhibit a seasonal variation, increasing during the winter months. The increases of both latencies during the winter were of similar magnitude, indicating that there were no significant changes in intertectal delay. The seasonal alteration in contralateral latency was not affected by dark-rearing and was affected to only a minor extent by a week-long alteration of ambient temperature.     

Contrasting behavioral responses to cold temperatures by two marine fish species during their pelagic juvenile interval

Motor activity of juvenile walleye pollock, Theragra chalcogramma, and sablefish, Anoplopoma fimbria, was monitored in the laboratory during high and low light levels under a changing temperature regime over a 5 d period. Water temperatures were ambient (12 °C) for the first 24 h of observation, rapidly lowered to 3 °C for the next 48 h, then raised back to 12 °C for the final 48 h. We hypothesized that the fishes' behavior would either follow a simple bioenergetic response of lowered activity associated with reduced metabolic rates at the colder temperature, or an avoidance response, with increased activity at decreased temperatures. Results for walleye pollock were consistent with a bioenergetic response, with activity decreasing in the presence of cold water under both high and low light levels, then returning to initial levels when temperatures increased. The response of sablefish, in contrast, indicated avoidance of cold temperatures, depending on light level. During high light, when sablefish were typically highly active, cold water induced a slight but insignificant decrease in activity. At low light, however, the presence of cold water caused a marked increase in sablefish movement through the experimental tanks, with a seven fold increase in the measured index of activity. When water temperatures were raised back to 12 °C, sablefish activity at low light returned to its normal, minimal level. The sharp increase in activity of sablefish in cold water, followed by a decrease in activity when the temperature was raised to pre-test levels, is suggestive of an avoidance response. The contrasting responses of the two species to thermal changes are consistent with their separate life history patterns and natural distributions.     

Reduced Pain and Inflammation in Juvenile and Adult Rats Fed a Ketogenic Diet

The ketogenic diet is a high-fat, low-carbohydrate regimen that forces ketone-based rather than glucose-based cellular metabolism. Clinically, maintenance on a ketogenic diet has been proven effective in treating pediatric epilepsy and type II diabetes, and recent basic research provides evidence that ketogenic strategies offer promise in reducing brain injury. Cellular mechanisms hypothesized to be mobilized by ketone metabolism and underlying the success of ketogenic diet therapy, such as reduced reactive oxygen species and increased central adenosine, suggest that the ketolytic metabolism induced by the diet could reduce pain and inflammation. To test the effects of a ketone-based metabolism on pain and inflammation directly, we fed juvenile and adult rats a control diet (standard rodent chow) or ketogenic diet (79% fat) ad libitum for 3–4 weeks. We then quantified hindpaw thermal nociception as a pain measure and complete Freund''s adjuvant-induced local hindpaw swelling and plasma extravasation (fluid movement from the vasculature) as inflammation measures. Independent of age, maintenance on a ketogenic diet reduced the peripheral inflammatory response significantly as measured by paw swelling and plasma extravasation. The ketogenic diet also induced significant thermal hypoalgesia independent of age, shown by increased hindpaw withdrawal latency in the hotplate nociception test. Anti-inflammatory and hypoalgesic diet effects were generally more robust in juveniles. The ketogenic diet elevated plasma ketones similarly in both age groups, but caused slowed body growth only in juveniles. These data suggest that applying a ketogenic diet or exploiting cellular mechanisms associated with ketone-based metabolism offers new therapeutic opportunities for controlling pain and peripheral inflammation, and that such a metabolic strategy may offer significant benefits for children and adults.