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.     

A simple cell labeling technique by means of lectins linked to fluorochromes for the detection of cells on tissue sections

Summary— We designed a protocol for cell labeling with the lectin wheat germ agglutinin (WGA) linked to the fluorochrome tetramethyl-rhodamine isothiocyanate (TRITC) for effective detection of the B16F10 melanoma and Lewis lung carcinoma (LLc) cells on pulmonary histological sections from C57BL6; mice. We have also determined a suitable concentration of WGA-TRITC (10 μg/ml), which leads to a very intense and homogeneous labeling of the cells, as it avoids cell clumping due to the presence of the lectin WGA. In order to determine to what extent the method affects these tumor cells, we have studied some important aspects related to their metastatic behavior, taking into account three parameters: a) viability and rate of proliferation of the cells cultured in vitro; b) percentage of animals C57BL6 mice) bearing metastasis 15 days after intravenous inoculation with 10⁵ B16F10 or LLc cells; and c) pattern of distribution of tumor foci in lung. There were no significant differences in these three parameters between the WGA-TRITC labeled-cells compared to the cultures of non-labeled cells in either of the cell lines (B16F10, LLc). Thus, we conclude that B16F10 and LLc tumor cells can be labeled following the protocol set-up in our study, as it allows these cells to be neatly identified on tissue sections and it causes no important physiological changes in the cells, with regard to metastatic behavior. These points make this technique very suitable for the detection of B16F10 and LLc cells on histological sections in studying their behavior during the first stages of the metastatic process.     

Elemental distribution in striated muscle and the effects of hypertonicity: Electron probe analysis of cryo sections

A method of rapid freezing in supercooled Freon 22 (monochlorodifluoromethane) followed by cryoultramicrotomy is described and shown to yield ultrathin sections in which both the cellular ultrastructure and the distribution of diffusible ions across the cell membrane are preserved and intracellular compartmentalization of diffusabler ions can be quantitated. Quantitative electron probe analysis (Shuman, H., A.V. Somlyo, and A.P. Somlyo. 1976. Ultramicros. 1:317-339.) of freeze-dried ultrathin cryto sections was found to provide a valid measure of the composition of cells and cellular organelles and was used to determine the ionic composition of the in situ terminal cisternae of the sarcoplasmic reticulum (SR), the distribution of CI in skeletal muscle, and the effects of hypertonic solutions on the subcellular composition if striated muscle. There was no evidence of sequestered CI in the terminal cisternae of resting muscles, although calcium (66mmol/kg dry wt +/- 4.6 SE) was detected. The values of [C1](i) determined with small (50-100 nm) diameter probes over cytoplasm excluding organelles over nuclei or terminal cisternae were not significantly different. Mitochondria partially excluded C1, with a cytoplasmic/ mitochondrial Ci ratio of 2.4 +/- 0.88 SD. The elemental concentrations (mmol/kg dry wt +/- SD) of muscle fibers measured with 0.5-9-μm diameter electron probes in normal frog striated muscle were: P, 302 +/- 4.3; S, 189 +/- 2.9;C1, 24 +/- 1.1;K, 404 +/- 4.3, and Mg, 39 +/- 2.1. It is concluded that: (a) in normal muscle the "excess CI" measured with previous bulk chemical analyses and flux studies is not compartmentalized in the SR or in other cellular organelles, and (b) the cytoplasmic C1 in low [K](0) solutions exceeds that predicted by a passive electrochemical distribution. Hypertonic 2.2 X NaCl, 2.5 X sucrose, or 2.2 X Na isethionate produced: (a) swollen vacuoles, frequently paired, adjacent to the Z lines and containing significantly higher than cytoplasmic concentrations of Na and Cl or S (isethionate), but no detectable Ca, and (b) granules of Ca, Mg, and P = approximately (6 Ca + 1 Mg)/6P in the longitudinal SR. It is concluded that hypertonicity produces compartmentalized domains of extracellular solutes within the muscle fibers and translocates Ca into the longitudinal tubules.     

Growth and derived life-history characteristics of the Brazilian electric ray Narcine brasiliensis

The majority of batoids are listed as Threatened (20.4%) or Data Deficient (41%) by the IUCN Red List. A key challenge to assessing Data-Deficient species is obtaining estimates of key life-history characteristics. Here, a Bayesian approach was used to estimate derived life-history characteristics from a growth model applied to the Data-Deficient Brazilian electric ray Narcine brasiliensis. The age of 170 specimens (107 females, 63 males) was estimated from vertebral centra, and total length, disc width, total weight and birth size were used in a joint estimation of sex-specific length-weight models and two-dimensional von Bertalanffy growth models. Estimates of age at length zero, age at maturity, longevity and mortality at age were derived simultaneously. The Bayesian joint modelling approach was robust to small sample sizes by adding a likelihood to constrain L₀ and sharing parameters, such as Brody growth coefficient between length measurements. The median growth parameter estimates were a shared L₀ = 38.8 mm, female L_∞ = 515 mm, 𝑘 = 0.125 and male L_∞ = 387 mm, 𝑘 = 0.194. Age at maturity was estimated to be 7.40–7.49 years for females and 4.45–4.47 years for males, whereas longevity was 22.5–22.6 years for females and 14.2 years for males depending on length measurement. Age-1 natural mortality was estimated to be 0.199–0.207 for females and 0.211–0.213 for males. The derived life-history characteristics indicate N. brasiliensis is earlier maturing, but slower growing relative to other Torpediniformes. These characteristics along with the species’ endemism to southern Brazil and high by-catch rates indicate that one of the IUCN Red List threatened categories may be more appropriate for the currently Data-Deficient status. The Bayesian approach used for N. brasiliensis can prove useful for utilizing limited age-growth data in other Data-Deficient batoid species to inform necessary life characteristics for conservation and management.     

Plant virus infection development as affected by heavy metal stress

The work was focused on the investigation of possible dependencies between the development of viral infection in plants and the presence of high heavy metal concentrations in soil. Field experiments have been conducted in order to study the development of systemic tobacco mosaic virus (TMV) infection in Lycopersicon esculentum L. cv. Miliana plants under effect of separate salts of heavy metals Cu, Zn and Pb deposited in soil. As it is shown, simultaneous effect of viral infection and heavy metals in tenfold maximum permissible concentration leads to decrease of total chlorophyll content in experiment plants mainly due to the degradation of chlorophyll a. The reduction of chlorophyll concentration under the combined influence of both stress factors was more serious comparing to the separate effect of every single factor. Plants' treatment with toxic concentrations of lead and zinc leaded to slight delay in the development of systemic TMV infection together with more than twofold increase of virus content in plants that may be an evidence of synergism between these heavy metal's and virus' effects. Contrary, copper although decreased total chlorophyll content but showed protective properties and significantly reduced amount of virus in plants.     

Maternal postnatal high-fat diet,rather than gestational diet,affects morphology and mTOR pathway in skeletal muscle of weaning rat

The positive regulation of insulin pathway in skeletal muscle results in increased activity of the mammalian target of rapamycin (mTOR), a positive effector of mRNA translation rate and protein synthesis. Studies that assess the activity of this protein in response to chronic high-fat diet (HFD) are scarce and controversial, and to date, there are no studies evaluating the mTOR pathway in infants exposed to gestational and postgestational HFD. This study investigated the effect of maternal HFD on skeletal muscle morphology and on phosphorylation of proteins that comprise the intracellular mTOR signaling pathway in soleus muscle of offspring at weaning. For this purpose, 10 days prior to conception, 39 female Wistar rats were randomly assigned to either control diet (CTL) or HFD. Later, rats were distributed into four groups according to gestational and postpregnancy diet: CTL/CTL (n=10), CTL/HF (n=11), HF/HF (n=10) and HF/CTL (n=8). After 21 days of lactation, pups were killed, and blood samples and soleus and gastrocnemius skeletal muscle were collected for analysis. We observed an influence of maternal postgestational diet, rather than gestational diet, in promoting an obese phenotype, characterized by body fat accumulation, insulin resistance and high serum leptin, glucose, triglycerides and cholesterol levels (P<.05). We have also detected alterations on skeletal muscle morphology — with reduced myofiber density — and impairment on S6 kinase 1 and 4E binding protein-1 phosphorylation (P<.05). These results emphasize the importance of maternal diet during lactation on muscle morphology and on physiological adaptations of infant rats.