A silver stain for the detection of nanogram amounts of tRNA following two-dimensional electrophoresis

Three ultrasensitive protein silver-staining methods have been compared with respect to the detection of tRNA in polyacrylamide gels. The method of Sammons (D. W. Sammons, L.D. Adams, and E.E. Nishizawa (1981) Electrophoresis 2, 135-141) has been shown to have remarkable sensitivity, with a detection limit of 0.3 ng tRNA/mm2, allowing the two-dimensional fractionation of submicrogram amounts of bulk tRNA. The application of this technique to developmental and differentiation problems and other areas where the amounts of nonradioactive tRNA available are limited is anticipated.     

Mitogenomic analysis of Montipora cactus and Anacropora matthai (cnidaria; scleractinia; acroporidae) indicates an unequal rate of mitochondrial evolution among Acroporidae corals

The complete nucleotide sequence of the mitochondrial (mt) genome was determined for specimens of the coral species Montipora cactus (Bernard 1897) and Anacropora matthai (Pillai 1973), representing two morphologically distinct genera of the family Acroporidae. These sequences were compared with the published mt genome sequence for the confamilial species, Acropora tenuis (Dana 1846). The size of the mt genome was 17,887 bp and 17,888 bp for M. cactus and A. matthai. Gene content and organization was found to be very similar among the three Acroporidae mt genomes with a group I intron occurring in the NADH dehyrogenase 5 (nad5) gene. The intergenic regions were also similar in length among the three corals. The control region located between the small ribosomal RNA (ms) and the cytochrome oxidase 3 (cox3) gene was significantly smaller in M. cactus and A. matthai (both 627 bp) than in A. tenuis (1086 bp). Only one set of repeated sequences was identified at the 3′-end of the control regions in M. cactus and A. matthai. A lack of the abundant repetitive elements which have been reported for A. tenuis, accounts for the relatively short control regions in M. cactus and A. matthai. Pairwise distances and relative rate analyses of 13 protein coding genes, the group I intron and the largest intergenic region, igr3, revealed significant differences in the rate of molecular evolution of the mt genome among the three species, with an extremely slow rate being seen between Montipora and Anacropora. It is concluded that rapid mt genome evolution is taking place in genus Acropora relative to the confamilial genera Montipora and Anacropora although all are within the relatively slow range thought to be typical of Anthozoa.     

Suppressive effects of soluble histocompatibility antigens on the in vitro generation of cytotoxic T cells to D-end alloantigens

Murine histocompatibility antigens were solubilized from the spleens and lungs of C57BL/6 (H-2b) animals with hypertonic salt (3 M KC1). Aggregate-free soluble antigens were incubated with nonadherent lymph node cells from BALB/c (H-2d) mice for 18 hr prior to their use as responder cells in the mixed-lymphocyte reaction (MLR). It was found that the generation of cytotoxic cells was suppressed while the proliferative response was not affected. The observed suppression was not due to a shift in the kinetics of the generation of cytotoxicity as determined throughout a 10-day culture period. The suppression was specific in that the response in MLR to unrelated H-2f stimulator cells and the subsequent generation of cytotoxic cells were unchanged. Using various H-2 recombinant strains as target cells in the assay of cell-mediated lympholysis, suppression of cytotoxicity was observed when the D end, but not the K end, was shared with the C57BL/6 strain from which the antigens were derived.     

Integrated palaeoecological and historical data in the service of fine-resolution land use and ecological change assessment during the last 1000 years in Rõuge, southern Estonia

Aims Our aim is to reconstruct decadal scale development of historical landscapes during the last 1000 years by means of fossil pollen analysis of annually laminated lake sediments, and detailed historical maps and documents. Location Lake Rõuge Tõugjärv (Estonia), a small lake with annually laminated lake sediments situated in a dense prehistoric setting. Methods The chronology of the palaeodata is based on the annual laminations supported by AMS ¹⁴C and ²¹⁰Pb dating and ¹³⁷Cs, ²⁴¹Am, and spheroidal carbonaceous particle marker horizons. The time‐scale and resolution allows fine sampling (the pollen samples generally comprise 3.5 years) and vegetation change reconstruction. Relevant source area of pollen (RSAP) of the lake was estimated, and the statistical zonation, rate of change, palynological richness, and DCA and PCA ordinations were generated on the basis of the pollen data. The historical calibration data set (maps, numerical information on population, domestic stock, farmland division, etc.) is based on archival material preserved in the Estonian Historical Archives. Results The topmost part (0–180 cm) of the sediment column of Lake Rõuge Tõugjärv, covering the last 1000 years, is visibly laminated carbonaceous gyttja. The varve chronology extends from ad 2000 to ad 1339, with a cumulative ± 9‐year error estimate. Beyond this the chronology is extrapolated using the ¹⁴C date and varve age–depth estimations. The simulation of the RSAP of Lake Tõugjärv shows that the major portion of the pollen loading originating from local vegetation is derived from plants growing within 2000 m of the sampling site. The pollen record divides into five statistically significant subgroups, which fall on the PCA plot into three clusters reflecting the general openness–closedness of the landscape. During the period between ad 1000 and 1200 (RT 1) the Rõuge area was generally wooded with birch, spruce and pine forests. The advancement of extensive farming gradually opened up the landscape between ad 1200 and 1650 (RT 2 and RT 3). The maximum openness of the landscape was reached between ad 1650 and 1875 (RT 4), with the most open period in the late eighteenth century. Historical maps from 1684 and 1870–99 and available quantitative data on population, domestic stock, farmland division, etc. show the same trend. The pollen data covering the last 125 years, and maps from 1935 and 1995, show the reduction of arable land in RSAP of the lake under investigation and the reduction of open land to an extent comparable with the end of the seventeenth century. Main conclusions The formation and development of the cultural landscape at Rõuge over the last 1000 years is characterized by rapid changes in floristic richness and rates of vegetation change attributed to certain historic processes in the RSAP. Five phases of landscape and social development are clearly distinguished during the last 1000 years. The decadal scale vegetation response to human‐induced forcing agrees with historical maps and documents and could be used for past landscapes prior to the period with solid historical data.     

Don C. Ohadike: The Man, his Intellectual Legacy and African Historiography

It is difficult to completely understand the life history of an intellectual excluding an understanding of his family upbringing and formative years. Family upbringing and childhood environment, often the less known part of a life history, play crucial roles in shaping the ideas and values individuals espouse in their adult life. Notwithstanding, this paper is not concerned with Don C. Ohadike’s childhood. It rather focuses on the professional career of our able historian – that is the part of his life as revealed by his most outstanding published writings. Ohadike’s published works contain a wellspring of idioms that tell much about his values, quality of mind, and his mission as an African historian. Ohadike was a humanist, an African patriot, and a nationalist crusader. His entire philosophy centered on safeguarding his African identity in an emergent world of cultural imperialism. The funds for this research were provided by a NEH-funded fellowship at the Schomburg Center, New York in the Spring of 2007. I owe a lot of gratitude to Professor John McLeod and Dean Blaine Hudson for granting me the extra incentives to pursue my research in New York. While all errors and misinterpretations are mine, I wish to thank the editors and anonymous reviewers for Journal of Dialectical Anthropology for their perspective comments and suggestions on earlier drafts of this paper.     

Ecology and biogeography of plant communities associated with the post Plio-Pleistocene relict Rhododendron ponticum subsp. baeticum in southern Spain

Aims Rhododendron ponticum L. is reputed to be a post Plio‐Pleistocene relict plant species with a disjunct distribution that comprises the Iberian Peninsula to the west and the Euxinian region plus some restricted Mediterranean areas to the east. We analysed the ecological range (of subsp. baeticum) in the western area (Aljibe Mountains, north of the Strait of Gibraltar) to understand the factors determining the present area limitation. Location Sierra del Aljibe, north of the Strait of Gibraltar (Iberian Peninsula). Methods We selected 20 riparian sites where R. ponticum is common, and compiled data on the ecological diversity of associated woody species and ferns. We established a 500‐m main transect in each site, along the stream or river course, in which we placed five 20‐m‐long plots at regular intervals. We recorded physiographic habitat features, woody plants and fern abundance, and the number of R. ponticum individuals. Results Rhododendron ponticum in southern Spain is restricted to riparian forests in acidic soils (pH 4.0–6.4), and is mainly found on the banks of inclined and enclosed streams. In our inventory we recorded 59 woody taxa and 12 ferns, with R. ponticum being the dominant species of the understorey (mean abundance 78.6%). The communities are characterized by a high incidence of the humid warm temperate element, both in number of species (18.8 ± 3.7 per site) and abundance; meanwhile, the presence of the modern Mediterranean element (mean number of species 3.4 ± 3.8 per site) appears to be favoured by disturbance. These ecological–historical groups of taxa also show distinct patterns of typological habit, frequency of endemism, infrageneric diversity and geographical range. Populations of R. ponticum are characterized by a very variable density of seedlings in many sites, and the virtual lack of juveniles. Main conclusions Riparian forests of the Aljibe Mountains constitute a refuge for R. ponticum where the species persists, but populations appear to be in decline. The narrow ecological range of R. ponticum in the area strongly contrasts with its wide amplitude in the eastern natural area, mainly the Euxinian region, where R. ponticum probably finds better conditions due to the environmental heterogeneity of the region, and the lack of a hot dry season.     

Characterization of a light-dependent glutamate synthase activity in Chlamydomonas reinhardtii

Photosynthetically active vesicles prepared from Chlamydomonas reinhardtii retained a light-dependent glutamate synthase activity which was highly specific for 2-oxoglutarate (K_m=2.1 mM) and L-glutamine (K_m=0.9 mM) as amido group acceptor and donor respectively. This activity was inhibited by azaserine, p-hydroxymercuribenzoate and 3-(p-chlorophenyl)-1,1-dimethyl urea.Light-dependent synthesis of glutamate was also obtained by coupling Chlamydomonas photosynthetic particles to purified ferredoxin-glutamate synthase, using ascorbate and 2,6-dichlorophenol-indophenol as electron donor. This system was also specific for 2-oxoglutarate (K_m=1 mM) and L-glutamine (K_m=0.8 mM) as substrates, and was stimulated by dithioerythritol. Azaserine and p-hydroxymercuribenzoate, but not 3-(p-chlorophenyl)-1,1-dimethyl urea, inhibited the reconstituted activity; high concentrations of 2-oxoglutarate were inhibitory.Abbreviations A Absorbance - CCP p-Trichlorometoxi-carbonylcyanide-phenylhydrazone - Chl Chlorophyll - CMU 3-(p-Chlorophenyl)-1,1-dimethyl urea - DPIP 2,6-Dichlorophenol-indophenol - DTE Dithioerythritol - MSX L-Methionine, D-L, sulfoximine - MV Methyl viologen