A 19-kb CpG Island Associated with Single-minded Gene 2 in Down Syndrome Chromosomal Region

To help in isolating the genes involved in Down syndrome, wesought CpG islands in 4 Mb cosmid/PAC contigs spanning mostof the 21q.22.2 band using seven rare cutting enzymes. A strikingfeature was observed upstream of hSIM2 where at least 41 rare-cuttingsites were clustered within a 20-kb region. To investigate thestructure of the cluster, a cosmid containing hSIM2 was submittedto shotgun sequencing. Sequence analysis revealed that the clusterwas a long CpG island extending 19, 128 nucleotides which includesin the first and second exons of hSIM2. Taken together withour observation in which the CpG islands were concentrated within1.2 Mb around hSIM2, we propose that this region functions asan R-band, and the cluster provides a unique element for markingof DNA for the spatial and temporal expression of the hSIM2locus.     

Skeletal muscle: A paradigm for testing principles of bioenergetics

Muscular activity converts chemical energy into useful work and metabolism restores muscle to its original state. This essay explores the organization and interactions of the regulatory system(s) which allow this energy balance to occur. The term energy balance is used in a biochemical rather than a thermodynamic sense—concerned not with deductions from the physical principles of thermodynamics, but rather with those enzymatic processes which nature evolved and which operate at remarkably fixed stoichiometry. Energy balance is a statement of conservation of energy put into biochemical observables.³¹P NMR spectroscopy is one of the most useful techniques for investigating these questions quantitatively under physiological conditionsin vivo. The author (1) describes the rules or principles of biochemical energy balance; (2) discusses sample results from human muscle to demonstrate its use in studying this class of questions; (3) presents a simple model of integrated cellular respiration to demonstrate its sufficiency to account for the main observations.     

Moral Darwinism: Ethical evidence for the descent of man

Could an ethical theory ever play a substantial evidential role in a scientific argument for an empirical hypothesis? InThe Descent of Man, Darwin includes an extended discussion of the nature of human morality, and the ethical theory which he sketches is not simply developed as an interesting ramification of his theory of evolution, but is used as a key part of his evidence for human descent from animal ancestors. Darwin must rebut the argument that, because of our moral nature, humans are essentially different in kind from other animals and so had to have had a different origin. I trace his causal story of how the moral sense could develop out of social instincts by evolutionary mechanisms of group selection, and show that the form of Utilitarianism he proposes involves a radical reduction of the standard of value to the concept of biological fitness. I argue that this causal analysis, although a weakness from a normative standpoint, is a strength when judged for its intended purpose as part of an evidential argument to confirm the hypothesis of human descent.     

Thein vitro effect of theophylline on 17α, 20ß-dihydroxy-4-pregnen-3-one-induced germinal vesicle breakdown in the catfish,Clarias batrachus

The effects of theophylline (a phosphodiesterase inhibitor) and cAMP on 17α, 20ß-dihydroxy-4-pregnen-3-one-induced germinal vesicle breakdown was investigatedin vitro in catfish (Clarias batrachus) oocytes. Folliculated oocytes incubated with 17α, 20ß-dihydroxy-4-pregnen-3-one at the concentration of 1 μg/ml induced 93.2 ± 2.23% germinal vesicle breakdown. When the oocytes were prestimulated with 17α,20ß-dihydroxy-4-pregnen-3-one for 6 h and then treated with different concentrations of theophylline, there was a significant drop in the frequency of germinal vesicle breakdown at the concentrations 2.0, 1.5 and 1.0 mM. However, theophylline was found to be incapable of inhibiting germinal vesicle breakdown at its lowest concentration (0.5 inM). In the time course study, significant inhibition of germinal vesicle breakdown was recorded when 1 mM theophylline was added up to 30 h of 17α,20ß-dihydroxy-4-pregnen-3-one Stimulation but the inhibitory effect of theophylline gradually (time dependent manner) declined if the stimulatory time of 17α,20ß-dihydroxy-4-pregnen-3-one was increased. A similar inhibition of germinal vesicle breakdown was also recorded with various concentrations of cAMP. Except 0.5 mM, all the higher concentrations of cAMP significantly inhibited 17α,20ß-dihydroxy-4-pregnen-3-one induced germinal vesicle breakdown.     

Phylogenetic relationships among transposon-like elements in human and primate DNA

A THE-1 sequence in intron 7 of the human dystrophin gene has been found to represent a new subfamily of THE-1 elements. The sequence is closely related to the MstII family of repetitive sequences and is more like single-copy sequences found in the galago genome than any other THE-1 sequence previously reported. This new THE-1 sequence has been compared with two other complete THE-1 sequences and three related long-terminal repeat elements that we have previously found in intron 7 of the dystrophin gene, and with members of the same family from elsewhere in the primate genome. Parsimony and deletion analysis show that the cluster of THE-1 sequences in intron 7 of the dystrophin gene has arisen from at least three individual insertion events, rather than from the insertion and duplication of a single progenitor sequence. Correspondence to: G.B. Petersen     

Adenosine effects upon insulin action on lipolysis and glucose transport in human adipocytes

The dose response effect of a new adenosine analogue, GR 79236 (N-[1S trans-2-hydroxycyclopentyl] adenosine) upon insulin sensitivity was examined in human adipocytes. The influence of adenosine upon insulin sensitivity for suppression of lipolysis and stimulation of glucose transport was examined. Removal of adenosine by use of adenosine deaminase stimulated lipolysis to the same extent as did 10^–9 M noradrenaline. GR79236 brought about dose dependent inhibition of lipolysis with half-maximal effect at 11.3±7.8×10^–9 M. When lipolysis was stimulated by noradrenaline alone the subsequent inhibition of lipolysis brought about by GR79236 was significantly greater than that of insulin. To examine adenosine effects on the insulin signalling pathway separately from those on lipolysis, the insulin sensitivity of glucose transport was examined. Removal of adenosine brought about a small but significant increase in the concentration of insulin required for half-maximal stimulation of glucose transport. Adenosine agonists offer promise as new agents for the modulation of metabolism in diabetes and other states of insulin resistance.     

Simulation and dynamic optimisation of animal cell culture

In animal cell culture, there are some 25 substrates that both have a significant effect on the culture performance and which can be measured with relative ease. A detailed dynamic simulation for such a culture has been produced and an optimisation policy that use this model to identify ideal media conditions has been developed. This paper describes an extension of that work to include the dynamic optimisation of cultures under fed-batch operation. Two different types of feeding policy were considered – in the first, discrete shots of feed were supplied, while in the second, feed was added continuously. Both policies offered significant improvements in the predicted productivity of the culture - up to 30% that of an experimentally optimisedbatch culture.     

Expression of the human milk protein β-casein in transgenic potato plants

A 1177 bp cDNA fragment encoding the human milk protein -casein was introduced into Solanum tuberosum cells under control of the auxin-inducible, bidirectional mannopine synthase mas12) promoters using Agrobacterium tumefaciens-mediated leaf disc transformation methods. Antibiotic-resistant plants were regenerated and transformants selected based on luciferase activity carried by the expression vector containing the human -casein cDNA. The presence of human -casein cDNA in the plant genome was detected by PCR and DNA hybridization experiments. Human -casein mRNA was identified in leaf tissues of transgenic plants by RT-PCR analysis. Human - casein was identified in auxin-induced leaf and tuber tissues of transformed potato plants by immunoprecipitation and immunoblot analysis. Human -casein produced in transgenic plants migrated in polyacrylamide gels as a single band with an approximate molecular mass of 30 kDa. Immunoblot experiments identified approximately 0.01% of the total soluble protein of transgenic potato leaf tissue as -casein. The above experiments demonstrate the expression of human milk - casein as part of an edible food plant. These findings open the way for reconstitution of human milk inedible plants for replacement of bovine milk in baby foods for general improvement of infant nutrition, and for prevention of gastric and intestinal diseases in children     

Conservation and management issues of Prespa National Park

The present article describes the human activities inthe area and their changes and trends, given that thecontemporary nature conservation key issues areclosely related to past and present socio-economic andcultural conditions, both within Prespa itself andoutside of it. The liberation of Prespa, a remote andrather socially isolated borderline area, from Ottoman occupation took place in 1912–13. Recentevents in its environmental history have been forestclearings and human depopulation in the Civil War(1944–49), irrigation system construction in the1960s, its discovery by ornithologists in the late1960s, the high emigration rates of 1960–1980, itsdesignation as a National Park in 1974, thedestructive development works of 1984–86 afterGreeces entry to the EEC and the conversion to intensivebean cultivation in the mid 1980s.The human population of ca 7000 at the start of thiscentury suffered an almost 80% decrease after theCivil War. A resettlement in the mid 1950s increasedthe population by 40% but in the 1970s emigrationreduced it to its previous size of around 1500, stillits present size. The extensive and diverse farmingsystems of previous centuries changed gradually afterthe construction of an irrigation network in the1960s, which twenty years later and along withinternational changes in production and economypatterns, made possible the present day domination ofa bean monoculture within irrigated croplands. Thiswas accompanied by increased energy inputs,mechanisation and the use of chemicals.Intensification of agriculture brought increasedincomes which attracted also livestock keepers,resulting in a 56% decrease in numbers of livestockin 1964–1993. Cattle are presently kept for meatproduction only. They graze on the mountains, and nolonger in the meadows around the lakeshore. A localshorthorn breed accounting for 93% of all cattle in1963 declined to less than 18% in 1993. Fishing aimedmainly at carp and Prespa bleak, continues today as inthe past to be a source of supplementary income.Eighty-six per cent of the – mainly oak and beech –forests are State owned and the rest belong to localmunicipalities. The forest area has not changedessentially in the last 40 years, but forest roadshave expanded. Forests provide mainly fuelwood forlocal needs and small quantities of industrial timber.No specific management is conducted to safeguard theconservation values of the forest, the majority ofwhich consists of even aged coppiced stands withlimited value for wildlife. Manufacturing never playedan important role for the economy of the area. Tourismhas increased in the last 20 years, concentratedmainly in summer and spring. Accommodation in thevillages is not yet satisfactory though it hasimproved enormously in recent years. The NationalParks infrastructure for visitors remains poor.Tourist pressure generally is not yet that high toimpact drastically the social and environmentalfabric. In places, hunting, angling and poaching cannegatively affect the target species. The constructionof the irrigation network in the eastern part of LakeMikri Prespa was the most important change to thelandscape and the habitats of Prespa the last 30years. It resulted in the embankment of free-flowingstreams, drainage of wet meadows and dramaticreduction of trees and hedges. In the last 15 years,the reedbeds, no longer burnt or mowed and assisted byincreased nutrient loadings from agricultural runoff,have expanded landward to occupy formerly open,periodically flooded, areas. The above changes inhabitats and land use have brought an impoverishmentof biodiversity which can be seen in the exterminationor the decrease in the numbers of certain plant andbird species.The problems preventing a better management of theNational Park and the perpetuation of its values areinappropriate legislation, the poor degree ofcoordination of the public services, the lack of botha specific National Park Service in Greece and amanagement authority for the Park and the hostileattitude of local people due to their limitedinformation and involvement. The scenic beauty, thewetlands and a number of rare habitats, the diversityof habitats, flora and fauna, the endemic life forms,the rare mammals and the colonial waterbirds, thelocal architecture and the cultural expressions suchas the Byzantine monuments, are the values of Prespathat must be preserved as a whole. The initialconservation efforts aimed at preserving biodiversitybut later it became apparent that not only are humansan inseparable part of the whole, but additionally,several of their extensive activities have contributedsubstantially to the rich biodiversity of the area.Through the prerequisites of keeping a balance betweenprimary and tertiary sector activities and ofrevitalizing the social fabric, the following are thekey management and conservation issues of today: anunequivocal agreement on the values to be preservedand their limits; the formation of a managementauthority; a new realistic protection and land usezoning; the diversification and extensification of allfarming activities; the restoration of the wet meadowhabitats; the ensuring of a high water level and agood water quality for both lakes through tri-lateralcooperation between Greece, Albania and FYROM; and theestablishment of a permanent environmental monitoringscheme.