In Situ Analysis of Na,K-ATPase α1- and α3-Isoform mRNAs in Aging Rat Hippocampus

Abstract: Age-related changes in the expression of Na,K-ATPase α1- and α3-isoform mRNAs were analyzed by in situ hybridization in the Fischer-344 rat hippocampus. Quantification of signal density with cRNA probes in rat hippocampus at 3 months of age showed (a) α1 content is 1.5 times higher in granule than in pyramidal cell layers, whereas α3 content shows the opposite ratio and (b) α3 label is found in large clusters related to mossy cells and basket cells and in medium clusters corresponding to interneurons within the dendritic fields of CA1–3. In the 24-month-old rats as compared with the young animals, the α1 signal is increased more than sevenfold in the dendritic fields and is not significantly changed in perikaryal layers. The α3 signal is reduced about threefold ( p < 0.0001, ANOVA, n = 6) in perikaryal layers, is almost completely absent over the interneurons, basket cells, and mossy cells, and is not significantly changed in dendritic fields. These data indicate age-related, cell- and isoform-specific alterations in pretranslational regulation of Na,K-ATPase α isoforms. The striking changes in the dendritic fields, mossy cells, and GABAergic basket cells and interneurons may constitute early and sensitive markers for age-related alterations in hippocampal function, before cell loss.     

Aminoadamantanes containing monoterpene-derived fragments as potent tyrosyl-DNA phosphodiesterase 1 inhibitors

The ability of a number of nitrogen-containing compounds that simultaneously carry the adamantane and monoterpene moieties to inhibit Tdp1, an important enzyme of the DNA repair system, is studied. Inhibition of this enzyme has the potential to overcome chemotherapeutic resistance of some tumor types. Compound (+)-3c synthesized from 1-aminoadamantane and (+)-myrtenal, and compound 4a produced from 2-aminoadamantane and citronellal were found to be most potent as they inhibited Tdp1 with IC₅₀ values of 6 and 3.5 µM, respectively. These compounds proved to have low cytotoxicity in colon HCT-116 and lung A-549 human tumor cell lines (CC₅₀ > 50 µM). It was demonstrated that compound 4a at 10 µM enhanced cytotoxicity of topotecan, a topoisomerase 1 poison in clinical use, against HCT-116 more than fivefold and to a lesser extent of 1.5 increase in potency for A-549.     

Olive oil history,production and by-product management

In this review paper, the history of the olive tree and the development of the main olive oil extraction systems employed in the past are presented. Furthermore, the management techniques employed from antiquity until today for the treatment of olive mill wastewater (OMW), which constitutes the most important by-product of olive oil production, are addressed and compared. Reference is also made to global olive oil production, the environmental impacts of the uncontrolled disposal of OMW, and to the legal framework concerning its management in the main olive oil producing countries. Other by-products of olive processing are also discussed.     

A general framework to describe the alteration of natural tree species composition as an indicator of forest naturalness

The alteration of natural tree species composition is defined as the deviation of the current tree species composition from that of the natural state. It can be used as a measure of human influence on forest vegetation, and thus as an indicator of the naturalness of forest vegetation. The aim of the study was to develop a standard procedure for estimating the alteration of natural tree species composition, to explain factors driving alteration and to examine its significance for susceptibility of forest stands to natural disturbances. The alteration of natural tree species composition was estimated for the Dinaric region (5556 km², Slovenia) by the Robič Index of Dissimilarity (RID), ranging from 0 (completely natural) to 100 (completely altered). The index was calculated on the compartment level (24 ha each on average) with data on current and potential natural forest vegetation. The influence of human activities on tree species alteration was examined by using topographic and accessibility variables. The susceptibility of forest stands to natural disturbances was analysed with data on sanitary felling. In the study area, the natural tree species composition of forest stands is moderately preserved; the average value of RID was 50.05, ranging from 1.76 to 100, and the coefficient of variation was 0.49. The alteration of the natural tree species composition of forest stands is primarily the result of forest management and past land use, conditioned either by topography or accessibility of forests. The degree of alteration of tree species composition decreased along the gradients of rockiness, inclination and elevation. A greater degree of alteration appeared on the slopes of intermediate and south facing aspects than on north facing slopes, and in areas that were closer to the forest edge. A higher level of alteration significantly increases the susceptibility of forest stands to natural disturbances. The procedure represents a novel approach in modelling the alteration (naturalness) of tree species composition of forest vegetation. It is applicable at different spatial scales and fosters an understanding of the patterns of tree species composition under the influence of human activity across forest landscapes.     

Allozyme polymorphisms, outerossing rates, and hybridization of South AmericanNothofagus

Electrophoretically detectable genetic variation was used to describe the genetic structure of three South American species ofNothofagus: the widespreadN. betuloides andN. dombeyi, and the geographically restrictedN. nitida. Although the widespread species possess higher levels of genetic variation, the three species have more genetic variation within than among populations. These results are consistent with the theoretical expectations for woody, presumably highly outerossed species with wind-borne seeds.Estimates of outcrossing rates from progeny arrays yielded slightly higher average t-values forN. nitida (1.158) andN. dombeyi (range 0.873–1.045) than forN. betuloide (0.878). Hierarchical analysis of population structure revealed values of F_IS and FIT that were positive and significantly different from zero at most loci and for each species. The levels of inbreeding detected by F-statistics indicate some degree of self-fertilization and/or population substructuring into discrete family groups. Reduced seed vagility and regeneration of natural stands after disturbance by a few remnant individuals would probably generate the recruitment of related seedlings underneath parent trees.The analysis of a putative hybrid population betweenN. nitida andN. betuloides indicated that individuals clearly segregated intonitida-like orbetuloides- like individuals. The reduced outcrossing rate ofbetuloides-like individuals from the hybrid site (t=0.585) is interpreted in concert with low pollen availability and the increased probability of selfing and/or hybridization inNothofagus.     

Detection of Plasmodium falciparum DNA in a microtitration plate-based hybridization test

A rapid DNA-test, depending on the affinity based hybrid collection principle, was developed for the detection of Plasmodium falciparum DNA from clinical specimens. In this method, hybridization takes place in solution and the hybrids are collected onto a solid phase for measurement. Two probes are used, one labelled with an affinity tag (biotin) and the other with a detectable label (32P). In the present test a single oligonucleotide complementary to a 21-base pair sequence which is highly repeated in the parasite genome served both as capture and detector probe. The test is a 2-h hybridization performed in streptavidin coated microtitration plate wells, onto which the labelled hybrids simultaneously bind. The sensitivity of the assay with a crude erythrocyte lysate specimen was 1.6 x 10(9) repeat units corresponding to about 160 parasites in one microliter blood. The results allowed quantification of the repeat sequences and thus estimation of the degree of parasitemia in clinical specimens.     

利用rpoB基因芯片技术进行快速分枝杆菌菌种鉴定

利用rpoB基因芯片技术快速进行分枝杆菌菌种鉴定。以分枝杆菌rpoB基因编码序列为靶基因, 用基因芯片技术检测21种分枝杆菌标准株；8种其它细菌标准株；126株临床分离株。分枝杆菌与其它细菌标准株经PCR扩增后, 分枝杆菌标准株均扩增出360 bp DNA片段, 在其它细菌中, 除甲型溶血性链球菌和假白喉棒状杆菌出现同样片段外, 其它细菌均未见扩增。21种寡核苷酸探针除海分枝杆菌与偶然分枝杆菌的探针有交叉杂交外, 其余均为特异性杂交。对126株临床分离株进行鉴定, 89株为结核分枝杆菌, 占70.6％(89/126), 非结核分枝杆菌(NTM)占9.2％(9/98)。应用rpoB基因芯片技术鉴定分枝杆菌菌种, 是一种快速、准确的方法, 具有较高的临床应用价值。     

Selection on floral traits through male fertility in a natural plant population

Most studies on selection in plants estimate female fitness components and neglect male mating success, although the latter might also be fundamental to understand adaptive evolution. Information from molecular genetic markers can be used to assess determinants of male mating success through parentage analyses. We estimated paternal selection gradients on floral traits in a large natural population of the herb Mimulus guttatus using a paternity probability model and maximum likelihood methods. This analysis revealed more significant selection gradients than a previous analysis based on regression of estimated male fertilities on floral traits. There were differences between results of univariate and multivariate analyses most likely due to the underlying covariance structure of the traits. Multivariate analysis, which corrects for the covariance structure of the traits, indicated that male mating success declined with distance from and depended on the direction to the mother plants. Moreover, there was directional selection for plants with fewer open flowers which have smaller corollas, a smaller anther–stigma separation, more red dots on the corolla and a larger fluctuating asymmetry therein. For most of these traits, however, there was also stabilizing selection indicating that there are intermediate optima for these traits. The large number of significant selection gradients in this study shows that even in relatively large natural populations where not all males can be sampled, it is possible to detect significant paternal selection gradients, and that such studies can give us valuable information required to better understand adaptive plant evolution. Second affiliation for Mark Van Kleunen is temporary until February 2007.