Exploring the functional significance of forest diversity: A new long-term experiment with temperate tree species (BIOTREE)

Effects of biodiversity on ecosystem functioning have been mainly studied in experiments that artificially create gradients in grassland plant diversity. Woody species were largely excluded from these early experiments, despite the ecological and socioeconomic importance of forest ecosystems. We discuss conceptual aspects of mechanistically driven research on the biodiversity–ecosystem functioning relationship in forests, including the comparison of scientific approaches like ‘observational studies’, ‘removal experiments’, and ‘synthetic-assemblage experiments’. We give a short overview on the differences between herbaceous and forest ecosystems, focusing on canopy characteristics, and the possibilities for individual versus population-based investigations.We present detailed information about the first large-scale, multisite and long-term biodiversity–ecosystem functioning experiment with tree species of temperate forests (BIOTREE – BIOdiversity and ecosystem processes in experimental TREE stands). At three sites of differing geology and local climate, we planted 200,000 saplings on a total area of 70 ha. At two sites, diversity gradients were established by varying the number of tree species (BIOTREE-SPECIES). At a third site, only functional diversity at a constant level of tree species richness was manipulated by selecting mixtures that differ in the functional trait values of the corresponding species (BIOTREE-FD). Additional experimental treatments at the subplot level include silvicultural management options, the addition of subdominant species, and the reduction of genetic diversity. Response variables focus on productivity, biogeochemical cycles and carbon sequestration, and resource use complementarity.We explore the use of different measures of functional diversity for a posteriori classifications of functional richness and their use in the analysis of our tree diversity experiment. The experiment is thought to provide a long-term research platform for a variety of scientific questions related to forest biodiversity and ecosystem processes.     

DNA ploidy abnormalities in rabbit preimplantation embryos are not increased by conditions associated with in vitro culture

Possible adverse effects of in vitro culture-associated physical factors were studied in 3- and 4-day-old rabbit embryos. Laboratory conditions were mimicked by exposure to visible light (320–740 nm, 1600 lx) or decreased temperature (22 ± 1°C). Embryos were exposed for a 24-hr period followed by either immediate evaluation or an additional 24 hr of standard in vitro culture (darkness, 37°C) and evaluation thereafter. Effects were assayed by cytophotometric measurement of the DNA content in Feulgen-stained cell nuclei and by cell number. The incidence of DNA aneuploid embryos and DNA aneuploid cell nuclei per embryo, as well as the average nuclear DNA content, was not significantly different between exposed embryos and controls. Both in vitro culture and reduced temperature caused a decrease in cell number. The temperature-induced cell number decrease was reversible within 24 hr after return to 37°C. These results demonstrate that physical factors associated with in vitro culture do not increase DNA ploidy abnormalities in cultured preimplantation embryos. Mol. Reprod. Dev. 50:30–34, 1998. © 1998 Wiley-Liss, Inc.     

Two distinct models account for short and long deletions within sequence repeats in Escherichia coli.

In Escherichia coli, (GpC)n sequences cloned into plasmid DNA molecules are deletion-prone with the occurrence of both short (<2 bp) and long (>2 bp) deletion events. These repetitive tracts can be stabilized by interrupting the strict monotony of the repetition with a variant dinucleotide sequence. The stabilization of short deletion events that is mediated by the variant sequence is completely lost in E. coli mismatch repair-deficient strains. In contrast, this repair pathway has no influence on the frequency of occurrence of long deletion events, even in sequences containing the variant repeat. These results lead us to propose two distinct models to account for short and long deletions within repetitive sequences in E. coli. Furthermore, this study reveals that the deletions occur preferentially at the end of the repeat sequence that is distal with respect to the origin of replication.     

Structure of the replication fork in ultraviolet light-irradiated human cells.

The DNA extracted from xeroderma pigmentosum human fibroblasts previously irradiated with 12.5 J/m2 of UV light and pulse-labeled for 45 min with radioactive and (or) heavy precursors, was used to determine the structural characteristics of the replication fork. Density equilibrium centrifugation experiments showed that a fork moved 6 micrometer in 45 min and bypassed 3 pyrimidine dimers in both strands. The same length was covered in 15-20 min in control cells. The delay in irradiated cells was apparently due to pyrimidine dimers acting as temporary blocks to the fork movement. Evidence for this interpretation comes from kinetics of incorporation of [3H]thymidine into DNA, which show that the time necessary to attain a new stable level of DNA synthesis in irradiated cells is equivalent to that required for the replication fork to cover the interdimer distance in one strand. On the other hand, the action of S1 nuclease on DNA synthesized soon after irradiation gives rise to a bimodal distribution in neutral sucrose gradients, one peak corresponding to 43 X 10(6) daltons and the other to 3 X 10(6) daltons. These two DNA species are generated by the attack of the S1 nuclease on single-stranded regions associated with the replication fork. A possible explanation for these results is given by a model according to which there is a delayed bypass of the dimer in the leading strand and the appearance of gaps opposite pyrimidine dimers in the lagging strand, as a direct consequence of the discontinuous mode of DNA replication. In terms of the model, the DNA of 43 X 10(6) daltons corresponds to the leading strand, linked to the unreplicated branch of the forks, whereas the piece of 3 X 10(6) daltons is the intergap DNA coming from the lagging strand. Pulse and chase experiments reveal that the low molecular weight DNA grows in a pattern that suggests that more than one gap may be formed per replication fork.     

Transcription‐blocking DNA damage in aging: a mechanism for hormesis

Recent evidence from studies on DNA repair systems that are implicated in accelerated aging syndromes, have revealed a mechanism through which low levels of persistent damage might exert beneficial effects for both cancer prevention and longevity assurance. Beneficial effects of adaptive responses to low doses of insults that in higher concentrations show adverse effects are generally referred to as hormesis. There are numerous examples of hormetic effects ranging from mild stresses of irradiation to heat stress, hypergravity, pro‐oxidants, or food restriction. Although the notion of hormesis is supported by many observations in various organisms, at least two major caveats have thus far prevented the application of hormesis for disease prevention in humans. First, the very nature of hormesis using toxins as a treatment regimen harbors the inherent danger of detrimental consequences. Second, the molecular mechanisms through which insults might exert beneficial effects have thus far remained elusive. Here, I discuss a mechanistic basis for hormesis and its implications for cancer prevention and healthy aging.     

Englerin A Agonizes the TRPC4/C5 Cation Channels to Inhibit Tumor Cell Line Proliferation

Englerin A is a structurally unique natural product reported to selectively inhibit growth of renal cell carcinoma cell lines. A large scale phenotypic cell profiling experiment (CLiP) of englerin A on ¬over 500 well characterized cancer cell lines showed that englerin A inhibits growth of a subset of tumor cell lines from many lineages, not just renal cell carcinomas. Expression of the TRPC4 cation channel was the cell line feature that best correlated with sensitivity to englerin A, suggesting the hypothesis that TRPC4 is the efficacy target for englerin A. Genetic experiments demonstrate that TRPC4 expression is both necessary and sufficient for englerin A induced growth inhibition. Englerin A induces calcium influx and membrane depolarization in cells expressing high levels of TRPC4 or its close ortholog TRPC5. Electrophysiology experiments confirmed that englerin A is a TRPC4 agonist. Both the englerin A induced current and the englerin A induced growth inhibition can be blocked by the TRPC4/C5 inhibitor ML204. These experiments confirm that activation of TRPC4/C5 channels inhibits tumor cell line proliferation and confirms the TRPC4 target hypothesis generated by the cell line profiling. In selectivity assays englerin A weakly inhibits TRPA1, TRPV3/V4, and TRPM8 which suggests that englerin A may bind a common feature of TRP ion channels. In vivo experiments show that englerin A is lethal in rodents near doses needed to activate the TRPC4 channel. This toxicity suggests that englerin A itself is probably unsuitable for further drug development. However, since englerin A can be synthesized in the laboratory, it may be a useful chemical starting point to identify novel modulators of other TRP family channels.     

Organization and activation of behavior in quail: role of testosterone metabolism

In quail, the hypothalamus enzymatically transforms testosterone (T) into estradiol (E2), 5 alpha-dihydrotestosterone (5 alpha-DHT), and 5 beta-dihydrotestosterone (5 beta-DHT). During the embryonic life, the 5 beta-reductase activity is very high, which probably protects the brain of males from being behaviorally demasculinized by their endogenous T. 5 beta androstanes are inactive androgens. The decrease of 5 beta reductase with age during sexual maturation corresponds to a potentiation of the effects of T as shown by experiments that compared the effects of T and 5 alpha-DHT in adult and young quail. T metabolism is also involved in the activation of male behavior in the adult. T aromatization is probably essential for behavioral activation, but nonaromatizable androgens such as methyltrienolone, and to some extent 5 alpha-DHT, can also stimulate sexual behavior in castrates. These enzymatic activities show a clear neuroanatomical localization and are sexually dimorphic. Males produce more active metabolites (E2, 5 alpha-DHT) than females, which could explain the male's greater sensitivity to T treatments. It thus appears that T metabolism is involved in the differentiation and activation of behavior in quail.     

Nutrient dynamics in a tritrophic system of ants,aphids and beans

The interactions between ants (Lasius niger), aphids (Aphis fabae) and plants (Phaseolus vulgaris) were studied in a laboratory experiment with the following treatments: application of N‐fertilizer for plants, supply of mealworms or sugar solution or their combination as alternative food sources for ants. Three main questions were studied: (1) Do ants reduce tending to honeydew‐producing aphids when an alternative sugar or protein resource was available? (2) Is aphid predation/protein consumption by ants higher when additional carbon is offered to maintain the carbon/protein balance? (3) Does fertilizer treatment propagate in the food web? For the experimental analysis stable isotope techniques were applied. δ¹⁵N served as a marker for the pathway from plants to higher trophic levels. Low δ¹⁵N‐value of fertilizer spread from plant shoots to aphids and ants. To trace which sugar‐/protein source was consumed by ants, the different ¹³C/¹²C‐ratios of C₃‐ and C₄‐plants were used with aphids feeding on C₃‐plant material, while mealworm food and sugar solution originated from C₄‐plant material. Fertilizer application had no effect on biomasses of plants, consumers or microflora. Ant biomass was significantly higher when additional sugar solution was offered. Higher contents of ¹³C indicated a high incorporation rate of additional sugar. Additional protein had no effect on colony biomass and no increasing predation on aphids could be observed when carbon was in excess. However, due to the lack of queens and newly produced larvae, protein requirements of experimental colonies were lower than in natural systems. Ants positively affected aphid populations, but reduced tending, whilst having access to an alternative sugar resource. When sugar/protein was offered to ants, the host plant had an increased root/shoot ratio. This indicates that decreasing aboveground activity of ants could lead to reduced plant growth of aphid‐infested plants, presumably due to higher fungal attack on shoots.