The Holdridge life zones of the conterminous United States in relation to ecosystem mapping

Aim Our main goals were to develop a map of the life zones for the conterminous United States, based on the Holdridge Life Zone system, as a tool for ecosystem mapping, and to compare the map of Holdridge life zones with other global vegetation classification and mapping efforts. Location The area of interest is the forty-eight contiguous states of the United States. Methods We wrote a PERL program for determining life zones from climatic data and linked it to the image processing workbench (IPW). The inputs were annual precipitation (Pann), biotemperature (T_bio), sea-level biotemperature (T₀bio), and the frost line. The spatial resolution chosen for this study (2.5 arc-minute for classification, 4-km for mapping) was driven by the availability of current state-of-the-art, accurate and reliable precipitation data. We used the Precipitation-elevation Regressions on Independent Slopes Model, or PRISM, output for the contiguous United States downloaded from the Internet. The accepted standard data for air temperature surfaces were obtained from the Vegetation/Ecosystem Modelling and Analysis Project (VEMAP). This data set along with station data obtained from the National Climatic Data Center for the US, were used to develop all temperature surfaces at the same resolution as the Pann. Results The US contains thirty-eight life zones (34% of the world's life zones and 85% of the temperate ones) including one boreal, twelve cool temperate, twenty warm temperate, four subtropical, and one tropical. Seventy-four percent of the US falls in the ‘basal belt’, 18% is montane, 8% is subalpine, 1% is alpine, and < 0.1% is nival. The US ranges from superarid to superhumid, and the humid province is the largest (45% of the US). The most extensive life zone is the warm temperate moist forest, which covers 23% of the country. We compared the Holdridge life zone map with output from the BIOME model, Bailey's ecoregions, Küchler potential vegetation, and land cover, all aggregated to four cover classes. Despite differences in the goals and methods for all these classification systems, there was a very good to excellent agreement among them for forests but poor for grasslands, shrublands, and nonvegetated lands. Main conclusions We consider the life zone approach to have many strengths for ecosystem mapping because it is based on climatic driving factors of ecosystem processes and recognizes ecophysiological responses of plants; it is hierarchical and allows for the use of other mapping criteria at the association and successional levels of analysis; it can be expanded or contracted without losing functional continuity among levels of ecological complexity; it is a relatively simple system based on few empirical data; and it uses objective mapping criteria.     

Membrane topology of the colicin E1 channel using genetically encoded fluorescence

The membrane topology of the colicin E1 channel domain was studied by fluorescence resonance energy transfer (FRET). The FRET involved a genetically encoded fluorescent amino acid (coumarin) as the donor and a selectively labeled cysteine residue tethered with DABMI (4-(dimethylamino)phenylazophenyl-4'-maleimide) as the FRET acceptor. The fluorescent coumarin residue was incorporated into the protein via an orthogonal tRNA/aminoacyl-tRNA synthetase pair that allowed selective incorporation into any site within the colicin channel domain. Each variant harbored a stop (TAG) mutation for coumarin incorporation and a cysteine (TGT) mutation for DABMI attachment. Six interhelical distances within helices 1-6 were determined using FRET analysis for both the soluble and membrane-bound states. The FRET data showed large changes in the interhelical distances among helices 3-6 upon membrane association providing new insight into the membrane-bound structure of the channel domain. In general, the coumarin-DABMI FRET interhelical efficiencies decreased upon membrane binding, building upon the umbrella model for the colicin channel. A tentative model for the closed state of the channel domain was developed based on current and previously published FRET data. The model suggests circular arrangement of helices 1-7 in a clockwise direction from the extracellular side and membrane interfacial association of helices 1, 6, 7, and 10 around the central transmembrane hairpin formed by helices 8 and 9.     

Rapamycin improves social and stereotypic behavior abnormalities induced by pre-mitotic neuronal subset specific Pten deletion

The mechanistic target of rapamycin (mTOR) pathway is a signaling system integral to neural growth and migration. In both patients and rodent models, mutations to the phosphatase and tensin homolog gene (PTEN) on chromosome 10 results in hyperactivation of the mTOR pathway, as well as seizures, intellectual disabilities and autistic behaviors. Rapamycin, an inhibitor of mTOR, can reverse the epileptic phenotype of neural subset specific Pten knockout (NS-Pten KO) mice, but its impact on behavior is not known. To determine the behavioral effects of rapamycin, male and female NS-Pten KO and wildtype (WT) mice were assigned as controls or administered 10 mg/kg of rapamycin for 2 weeks followed by behavioral testing. Rapamycin improved social behavior in both genotypes and stereotypic behaviors in NS-Pten KO mice. Rapamycin treatment resulted in a reduction of several measures of activity in the open field test in both genotypes. Rapamycin did not reverse the reduced anxiety behavior in KO mice. These data show the potential clinical use of mTOR inhibitors by showing its administration can reduce the production of autistic-like behaviors in NS-Pten KO mice.     

Isolation of microcell hybrid clones containing retroviral vector insertions into specific human chromosomes.

We sought an efficient means to introduce specific human chromosomes into stable interspecific hybrid cells for applications in gene mapping and studies of gene regulation. A defective amphotropic retrovirus was used to insert the gene conferring G418 resistance (neo), a dominant selectable marker, into the chromosomes of diploid human fibroblasts, and the marked chromosomes were transferred to mouse recipient cells by microcell fusion. We recovered five microcell hybrid clones containing one or two intact human chromosomes which were identified by karyotype and marker analysis. Integration of the neo gene into a specific human chromosome in four hybrid clones was confirmed by segregation analysis or by in situ hybridization. We recovered four different human chromosomes into which the G418 resistance gene had integrated: human chromosomes 11, 14, 20, and 21. The high efficiency of retroviral vector transformation makes it possible to insert selectable markers into any mammalian chromosomes of interest.     

Cyclosporin A inhibits calcineurin (phosphatase 2B) and P-glycoprotein activity in Entamoeba histolytica

Cyclosporin A (CsA) inhibits the proliferation of several protozoan parasites through blocking the activity of calcineurin (Cn) or P-glycoproteins (Pgp). We report here, that inhibition of the proliferation of Entamoeba histolytica trophozoites, the causal agent of human amebiasis, is due to interference of the phosphatase activity of Cn, in a similar fashion to the effect of this immunosuppressive drug on T lymphocytes. The non-immunosuppressive CsA analog PSC-833, which binds Pgp without interfering the function of Cn, did not inhibit the proliferation of HM1:IMSS trophozoites. Moreover, phosphatase activity of amebic Cn, detected using the phosphopeptide RII, was drastically affected by incubation with CsA, but not with PSC-833. On the other hand, both drugs were also tested on clone C2 trophozoites, which grow in the presence of emetine due to over-expression of Pgp. The effect of CsA was similar to that observed on HM1:IMSS trophozoites, whereas PSC-833 only affected the proliferation and viability of clone C2 when the trophozoites were grown in the presence of 40 microM of emetine, suggesting an interference of the Pgp activity. This suggestion was confirmed by results from experiments of Pgp-dependent effux of rhodamine from pre-loaded trophozoites, in the presence of either of these drugs. Therefore, CsA inhibition of E. histolytica trophozoite proliferation is more likely due to Cn than Pgp activity inhibition.     

Mineral content of leaves from trees growing on serpentine soils under contrasting rainfall regimes in Puerto Rico

The interactions between water and soil nutrient availability in determining leaf nutritional composition and structural features were investigated in forests on serpentine in Maricao and Susua (Puerto Rico). These forests grow under contrasting rainfall regimes: Maricao is a wet forest located at altitudes above 500 m and receiving more than 2500 mm rainfall, while Susua is a humid forest located well below 500 m, with less than 1500 mm rainfall and a well defined dry season. Dominant tree species and soils were analysed for N, P, K, Ca, Mg and Ni. Soils can be differentiated according to their K content (higher in Maricao) and P contents (higher in Susua). Mature leaves of both forests have sclerophyllous characteristics as judged from the Specific Leaf Areas (<80 cm² g^-1) and low P contents. Leaf area development is strongly correlated with leaf N and P contents in both forests, but Maricao samples appear to be more limited by P availability. In concordance with soil values, the Susua leaf sample set has significantly higher contents of P, but lower contents of K when compared with the Maricao sample set. Analyses of soluble K, Ca, and Mg reveal strong physiological selectivity in the absorption of these cations. K/Ca and Ca/Mg ratios are markedly higher in the soluble leaf extracts than in the soil extracts. It seems that restriction to vegetation development in the serpentine areas investigated are more related to nutritional deficiencies and not to high contents of either Mg or Ni in the upper soil layers. Only two strong Ni accumulators were found, Cassine xylocarpa (1.2 mol Ni g^-1 dry mass or 70 g g^-1) from Susua, and Chionanthus domingensis (12.2 mol g^-1, or about 700 g g^-1) from Maricao. These species are not restricted to serpentine areas in Puerto Rico.     

Isolation of thymidine kinase-deficient rat hepatoma cells by selection with bromodeoxyuridine,Hoechst 33258, and visible light

The photosensitivity of bromodeoxyuridine (BrdU)-substituted cells is known to be markedly enhanced by the fluorochrome Hoechst 33258. Since the incorporation of BrdU into nucleic acids depends upon its prior phosphorylation via thymidine kinase (TK; EC 2.7.1.21), cells deficient in TK activity are refractory to photoinduced killing. These observations suggested that combined treatment with BrdU, Hoechst 33258, and visible light would constitute an efficient selective strategy for the recovery of TK^– mutant cells. In this report we describe a single-step selection protocol which reduced the survival of TK⁺ cells by a factor of 10⁵ without affecting the viability of TK^– mutants. This procedure was used to isolate H4IIEC3-derived rat hepatoma cells deficient in TK activity. The properties of several TK^– hepatoma clones are discussed.The valuable technical assistance of J. M. Courvall, F. R. Parker, and M. M. Smith is acknowledged. These studies were supported by grant GM26449 from the National Institutes of Health. A.M.K. was supported by a postdoctoral fellowship from the American Cancer Society, California Division. T.G.L. is a Leukemia Society of America postdoctoral fellow. R.E.K.F. is the recipient of an American Cancer Society Faculty Research Award.