Somatic embryogenesis in cell cultures of birch (Betula pendula Roth.)

Somatic embryogenesis was induced in cell cultures of birch (Betula pendula Roth.) derived from juvenile tissue of seed embryos and from mature leaf tissue. Embryos were formed in liquid and on solidified medium containing 2,4-dichlorophenoxyacetic acid (2,4-D) and 6-furfurylaminopurine (kinetin). Sometimes somatic embryos formed only after transfer to medium devoid of growth regulators. The embryos germinated on hormone-free medium and were potted in soil and grown in the greenhouse.Finnish Forest Research InstituteUniversity of Helsinki, School of Pharmacy;     

Morphological variation among clones of ulluco (Ullucus tuberosus,Basellaceae) collected in Southern Peru

Material of 229 clones of ulluco (Ullucus tuberosus), representing 10 accessions from markets in southern Peru and one from the wild in Bolivia, was analyzed with regard to morphological variation. The diploid chromosome number (2n = 24) was ascertained in 16 of the clones. Clones grown in different environments (e.g., long vs. short day) retained a more or less similar order of variation with respect to color of the tubers, stems, and leaves and the length/width ratio of the leaves. The shape of fully developed tubers also showed stable variation. Within a single experiment several other vegetative characters showed significant differences between accessions. Furthermore, many flower and inflorescence characters showed variation between accessions, some of them also indicating geographical variation between the Puno-Juliaca area and the Cuzco area. Considerable variation between the clones of an accession was evident, some accessions being clearly more variable than others. Our results show that ulluco is a crop plant that varies extensively even within a geographically limited area. Although repeatedly referred to in the literature as a purely asexual crop, ulluco was shown to be capable of sexual reproduction; this must have been and possibly still is an important source of genetic variation in the species.     

Computational models for the shuttling motion of the macrocycle in rotaxane-based molecular switches

The shuttling motion of a macrocycle in rotaxane-based molecular switching devices has been studied by computational density functional methods. In the test case, energy profiles corresponding to the dethreading process of different types of guest molecules in a cyclobis(paraquat-p-phenylene) host verified the experimental preference of the tetrathiafulvalene recognition site over the dioxynaphthalene site in a Stoddart-Heath type molecular device. Furthermore, modification of the redox state of either the macrocycle or the guest molecule resulted in considerable changes in the computational energy profiles, which can be utilized in explaining the behavior of the host-guest system. In order to study the effect of chemical oxidation/reduction in the guest molecule, we have investigated a prototypical shaft including two octahedral ruthenium complexes linked by a conjugated C₁₄ carbon chain, where the shuttling motion can be triggered by changing the electronic environment of the active complexes with ligand exchange reactions. The computational results also indicated effective communication between the macrocycle and the conjugated carbon chain, therefore showing the importance of non-covalent host-guest interactions in the control of the motion. Figure A computational model for a chemically controlled molecular switch     

Colonization routes of Pinus sylvestris inferred from distribution of mitochondrial DNA variation

Understanding the present-day distribution of molecular variation requires knowledge about the history of the species. Past colonization routes and locations of refugia of Scots pine (Pinus sylvestris) were inferred from variation in mitochondrial DNA in material collected from 37 populations located in countries within, and immediately adjacent to the continent of Europe. Two mitochondrial regions, nad1 intron (exon B/C) and nad7 intron 1, were included in the study. Differentiation in maternally inherited mitochondria was high (G _ST′ = 0.824). Two new haplotypes were found at the nad7 intron 1. The occurrence of a 5-bp indel variant was restricted to the Turkish Kalabak population and a 32 bp only found in Central, Eastern, and Northern Europe. The complete absence of the 32-bp indel from the Mediterranean peninsulas supports the view that coniferous forests existed outside these areas during the last glacial maximum, and these populations contributed to the subsequent colonization of the northern parts of Europe. P. sylvestris shares features of its glacial and postglacial history with two other northern, cold-tolerant tree species, Picea abies and Betula sp. These three species differ from many other European trees for which pollen core and molecular evidence indicate colonization from southern refugia after the last glacial period.     

Reconsideration of hydrophobic lipid distributions in lipoprotein particles

Lipoprotein particles are commonly known as micellar aggregates with hydrophobic lipids located within the core and amphipathic molecules in the surface. Using a new structural model for optimizing the distribution of hydrophobic lipids, namely triglyceride (TG) and cholesterol ester (CE) molecules, we reveal that particle size-dependent proportion of these 'core lipids' may locate in the surface of lipoprotein particles. The composition of the particles also strongly influences the actual molecular content of the surface. For example, in high-density lipoprotein (HDL) particles the percentage of CEs of all surface lipids is between 13% and 27% due to the high tendency of CEs to locate in the surface and the high concentration of CEs in the particles. Conversely, although the percentage of TG molecules in the surface of HDL particles is also high, approximately 60% as for CE, the percentage of TGs of all surface lipids is low, only up to 5%, because HDL particles have a low-TG concentration. These structural models provide an intuitive and coherent structural rationale for various metabolic cascades in lipoprotein metabolism with the catalytic enzyme action and molecular binding for transport proteins taking place at the surface of the particles.     

Chromosomal regions underlying noncoagulation of milk in Finnish Ayrshire cows

About 10% of Finnish Ayrshire cows produce noncoagulating milk, i.e., milk that does not form a curd in a standard 30-min testing time and is thus a poor raw material for cheese dairies. This phenomenon is associated with peak and midlactation, but some cows produce noncoagulating milk persistently. A genomewide scan under a selective DNA pooling method was carried out to locate genomic regions associated with the noncoagulation of milk. On the basis of the hypothesis of the same historical mutation, we pooled the data across sires. Before testing pools for homogeneity, allele intensities were corrected for PCR artifacts, i.e., shadow bands and differential amplification. Results indicating association were verified using daughter design and selective genotyping within families. Data consisted of 18 sire families with 477 genotyped daughters in total, i.e., 12% of each tail of the milk coagulation ability. Data were analyzed using interval mapping under maximum-likelihood and nonparametric methods. BMS1126 on chromosome 2 and BMS1355 on chromosome 18 were associated with noncoagulation of milk across families on an experimentwise 0.1% significance level. By scanning gene databases, we found two potential candidate genes: LOC538897, a nonspecific serine/threonine kinase on chromosome 2, and SIAT4B, a sialyltransferase catalyzing the last step of glycosylation of κ-casein on chromosome 18. Further studies to determine the role of the candidates in the noncoagulation of milk are clearly needed.     

Coleopteran diversity and abundance in different habitats near Kihansi waterfall, in the Udzungwa Mountains, Tanzania

Coleoptera diversity at the family level was investigated along the Kihansi gorge near a 700 m high waterfall system which will be taken to hydropower use, and the current river flow will be diverted due to dam construction. The coleopteran communities of three micro-habitats: spray zone, forest site and riverine site were compared by sweepnetting and pitfall trap methods. The highest Coleopteran family level diversity was found in the spray zone where the Shannon–Weaver index of diversity was 0.71 (forest site 0.31; riverine site 0.50). Coleopterans were most abundant in the forest site where 44% of all sampled individuals were found (spray zone 31%; riverine site 23%). Most of less frequently recorded families were found in the spray zone. Coleoptera families were found to be unequally partitioned in all three micro-habitats. The highest percentage similarity index (85%) was found between forest and riverine sites (spray/forest 76%; spray/riverine 79%). Distributions of abundances of coleopteran families were significantly different between all studied habitats. The study revealed that the spray generated by the waterfall provide a special micro-habitat for Coleoptera. It is suggested that conservation efforts and monitoring in the study area using selected taxonomic indicator Coleopteran groups should be carried out in order to help to adjust mitigation measures.     

Diffusion of KCl in an amylose film

A method has been developed measuring the diffusion coefficient of KCl in amylose films. The films were soaked in potassium chloride solutions, then immersed in pure water and conductivity measured as a function of time. Different concentrations of the soaking solution were used and the measurements were made at several temperatures. The diffusion coefficient of KCl was found to be independent of the soaking solution KCl concentration, but found to increase with increasing temperature. The diffusion coefficient values were about one quarter of those found in water and varied from 4.8×10⁻¹⁰ to 11×10⁻¹⁰ m² s⁻¹. The activation energy of diffusion was close to that found in water. Two values for the activation energy were obtained, 20.1 and 14 kJ mol⁻¹, indicating a change in the film structure at 45 °C. Amylose films swelled equally in KCl-solutions and water. The thickness of amylose films doubled and the increase in mass was 100–200% corresponding the decrease of amylose content from about 87 to 37%, when the conditions changed from normal humidity conditions to water.     

Association of cholesteryl ester transfer protein with HDL particles reduces its proteolytic inactivation by mast cell chymase

Human atherosclerotic intima contains mast cells that secrete the neutral protease chymase into the intimal fluid, which also contains HDL-modifying proteins, such as cholesteryl ester transfer protein (CETP), in addition to abundant amounts of nascent discoidal HDL particles. Here, we studied chymase-dependent degradation of a) CETP isolated from human plasma and b) CETP-HDL complexes as well as the functional consequences of such degradations. Incubation with chymase caused a rapid cleavage of CETP, yielding a specific proteolytic pattern with a concomitant reduction in its cholesteryl ester transfer activity. These chymase-dependent effects were attenuated after CETP was complexed with HDL. This attenuation was more effective when CETP was complexed with HDL(3) and HDL(2) than with discoidal reconstituted high density lipoprotein (rHDL). Conversely, rHDL, but not spherical HDLs, was protected in such CETP complexes against functional inactivation by chymase. Thus, in contrast to the complexes of CETP with spherical HDLs, the ability of the CETP-rHDL complexes to promote cholesterol efflux from macrophage foam cells remained unchanged, despite treatment with chymase. In summary, complexation of CETP and HDL modifies their resistance to proteolytic inactivation: spherical HDLs protect CETP, and CETP protects discoidal HDL. These results suggest that in inflamed atherosclerotic intima, CETP, via its complexation with HDL, has a novel protective role in early steps of reverse cholesterol transport.