Can the preservation of historical relicts permit the conservation of endangered plant species? The case of <Emphasis Type="Italic">Silene sennenii</Emphasis> (Caryophyllaceae)

Allozyme electrophoresis was used to measure levels and distribution of genetic diversity in the critically endangered (CR) Silene sennenii (Caryophyllaceae), a narrow endemic plant species found in northeastern Catalonia (Spain). At present, only 5 populations remain, containing no more than 5,000 individuals, which are subjected to several human pressures such as habitat fragmentation and land use changes. Interestingly, the largest population is located in and around a fortress built in the XVIII century, a circumstance which might facilitated its preservation until now. From the 21 satisfactorily interpreted loci, low levels of genetic variation were detected (P = 20.9, A = 1.31 and H _e = 0.071), which may be related to small population size, isolation, and fragmentation of extant populations. Moderate to high levels of inbreeding were also found, probably as consequence of the population’s genetic structuring. The conservation of the population located in the fortress would allow the preservation of all the alleles detected at species level; nevertheless, conservation of other populations, coupled with the inclusion of S. sennenii in the Spanish Catalogue of Endangered Species, would also be desirable in order to ensure the long-term survival of the species.     

Determination of fluoxetine, norfluoxetine and their enantiomers in rat plasma and brain samples by liquid chromatography with fluorescence detection

Fluoxetine (FLX) and norfluoxetine (NFLX) racemic mixtures were determined by reversed-phase liquid chromatography with fluorescence detection (lambda(exc)=227 nm, lambda(em)=305 nm). The calibration curves prepared from drug-free plasma and brain were linear in the range of 5-1000 ng ml(-1) and 100-40,000 ng g(-1) for doped samples, with detection limits of 3.2 and 2.1 ng ml(-1) in plasma and 31.5 and 26.1 ng g(-1) in brain tissue for FLX and NFLX, respectively. Enantiomer determination was carried out through normal phase HPLC-FD (lambda(exc)=224 nm, lambda(em)=336 nm) after precolumn chiral derivatization with R-1-(1-naphthyl)ethyl isocyanate. Standard curves also prepared in a drug-free matrix were linear for each enantiomer over the range of 2-1000 ng ml(-1) and 20-7000 ng g(-1) with detection limits for the four compounds ranging between 0.2 and 0.5 ng ml(-1) in plasma and between 3.0 and 8.2 ng g(-1) in brain tissue. In both methods the analytes were isolated from the biological matrix by a new solid-phase extraction procedure with recovery in plasma and brain over 90 and 87%, respectively. The repeatability of this extraction procedure was satisfactory within-day and between-day with CV<9.1%. This study also offered the opportunity to obtain an assessment of the potential relationships between the concentration of individual enantiomers of FLX and NFLX in plasma and brain tissue after chronic treatment with racemic FLX at a dose intended to mimic the human plasma concentration of FLX in standard clinical conditions, and therefore should make for more reliable extrapolation of neurochemical findings in other species.     

Differential intestinal mucosal protein expression in hypercholesterolemic mice fed a phytosterol-enriched diet

The molecular mechanisms involved in the phytosterol-induced decrease in intestinal cholesterol absorption remain unclear. Further, other biological properties such as immunomodulatory activity and protection against cancer have also been ascribed to these plant compounds. To gain insight into the mechanisms underlying phytosterol actions, we conducted a proteomic study in the intestinal mucosa of phytosterol-fed apolipoprotein E-deficient hypercholesterolemic (apoE-/-) mice. With respect to control-fed apoE-/- mice, nine differentially expressed proteins were identified in whole-enterocyte homogenates using 2-D DIGE and MALDI-TOF MS. These proteins are involved in plasma membrane stabilization, cytoskeleton assembly network, and cholesterol metabolism. Four of these proteins were selected for further study since they showed the highest abundance change or had a potential functional relationship with known effects of phytosterols. Annexin A2 (ANXA2) and beta-actin decrease and annexin A4 (ANXA4) and annexin A5 (ANXA5) increase were confirmed by Western blot analysis. Intestinal gene expression of ANXA2 and A5 and beta-actin was reduced, whereas that of ANXA4 was unchanged. The main results were retested in normocholesterolemic C57BL/6J mice. ANXA4 and ANXA5 protein upregulation and ANXA2 and beta-actin downregulation were reproduced in these animals. However, no changes in gene expression were found in C57BL/6J mice in either of the four proteins selected. ANXA2, A4, and A5 and beta-actin are proteins of special interest given their pleiotropic functions that include cholesterol-ester transport from caveolae, apoptosis, and anti-inflammatory properties. Therefore, the protein expression changes identified in this study might be involved in the biological effects of phytosterols.     

Monocyte chemoattractant protein-1 in obesity and type 2 diabetes. Insulin sensitivity study

Objective: Our goal was to test any association between human plasma circulating levels of monocyte chemoattractant protein‐1 (cMCP‐1) and insulin resistance and to compare monocyte chemoattractant protein‐1 (MCP‐1) adipose tissue gene expression and cMCP‐1 in relation with inflammatory markers. Research Methods and Procedures: cMCP‐1 was measured in n = 116 consecutive control male subjects to whom an insulin sensitivity (S_i) test was performed. Circulating levels of soluble CD14, soluble tumor necrosis factor receptor type 2 (sTNFR2), soluble interleukin‐6 (sIL‐6), and adiponectin also were measured. Subcutaneous adipose tissue samples were obtained from n = 107 non‐diabetic and type 2 diabetic subjects with different degrees of obesity. Real‐time polymerase chain reaction was used to measure gene expression of MCP‐1, CD68, tumor necrosis factor‐α (TNF‐α), and its receptor TNFR2. Results: In the S_i study, no independent effect of cMCP‐1 levels on insulin sensitivity was observed. In the expression study, in non‐diabetic subjects, MCP‐1 mRNA had a positive correlation with BMI (r = 0.407, p = 0.003), TNF‐α mRNA (r = 0.419, p = 0.002), and TNFR2 mRNA (r = 0.410, p = 0.003). In these subjects, cMCP‐1 was found to correlate with waist‐to‐hip ratio (r = 0.322, p = 0.048). In patients with type 2 diabetes, MCP‐1 mRNA was up‐regulated compared with non‐diabetic subjects. TNF‐α mRNA was found to independently contribute to MCP‐1 mRNA expression. In this group, CD68 mRNA was found to correlate with BMI (r = 0.455, p = 0.001). Discussion: cMCP‐1 is not associated with insulin sensitivity in apparently healthy men. TNF‐α is the inflammatory cytokine associated with MCP‐1 expression in subcutaneous adipose tissue.     

The relationship between relative weight and school attendance among elementary schoolchildren

Objective: To determine the relationship between relative weight and school attendance among elementary schoolchildren. Research Methods and Procedures: A total of 1069 fourth to sixth graders from nine elementary schools in the inner city of Philadelphia, PA, were part of an ongoing randomized control trial to assess prevention strategies for obesity. The mean rate of students eligible for free/reduced meals was 82.9 ± 11.5%. Weight was measured in the second semester of the academic year. Absentee data for the entire academic year were recorded by homeroom teachers. Participants were classified into relative weight categories described by the Institute of Medicine: underweight, normal‐weight, overweight, and obese. Results: ANOVA showed that overweight children were absent significantly more than normal‐weight children (12.2 ± 11.7 days vs.10.1 ± 10.5 days) (p < 0.05). Linear regression showed that the obese category remained a significant contributor to the number of days absent even after adjusting for age, race/ethnicity, and gender. Discussion: These data suggest that in addition to the medical and psychosocial consequences of being overweight, heavier children have greater risk for school absenteeism than their normal‐weight peers. As the rate of childhood obesity increases, parallel increases in school absenteeism should be expected.     

Temperature effect on tert-butyl alcohol (TBA) biodegradation kinetics in hyporheic zone soils

Background  

Remediation of tert-butyl alcohol (TBA) in subsurface waters should be taken into consideration at reformulated gasoline contaminated sites since it is a biodegradation intermediate of methyl tert-butyl ether (MTBE), ethyl tert-butyl ether (ETBE), and tert-butyl formate (TBF). The effect of temperature on TBA biodegradation has not been not been published in the literature.     

Population biology of <Emphasis Type="Italic">Kosteletzkya pentacarpos</Emphasis> (Malvaceae) in the Llobregat delta (Catalonia,NE of Spain)

Field experiments and wild population monitoring have been performed to study the population biology of the rare long-lived Kosteletzkya pentacarpos (Malvaceae) in the Llobregat delta (Catalonia, NE Spain). Field experiments explored the fate of seeds in soil at different depths, seedling emergence, and seedling survival, growth and flowering with and without canopy cover during the first 2 years of life. They also were used to ascertain the size-related pattern of seedling survivorship and flowering. Field data concerning mortality, growth and fecundity of adult plants were collected yearly in three wild populations for 7–9 years. In old adults (reproducing long before the beginning of the study), ANOVAR tests were performed to compare maximum diameter, total and fertile shoots, and viable seeds per plant between years and populations. New adults (starting flowering the first year of study or in subsequent years) were used to explore, using linear and polynomial regressions, the association of RGR and both total and fertile shoot production with (i) plant size (maximum basal diameter or its logarithm); (ii) plant age (years in adult stage); and (iii) plant age after removing the effect of size and year-to-year fluctuations. In this case, we examined the age-related pattern of the residuals obtained from the regressions with size and year. The study identified the following main demographic features of K. pentacarpos: (i) transient, shallow soil seed bank; (ii) shade tolerance of seedling emergence; (iii) canopy-facilitation of seedling survival and bolting during the first two years of life; (iv) size-related pattern for seedling survivorship but not flowering; (v) exclusive dependence on a fluctuating seed output for reproduction; (vi) rapid adult growth; and (vii) high adult longevity but (viii) rapid depletion of fecundity with age. Seed output was highly constrained by mining insects. The changing size-structure and the decreasing reproductive success of old adults in several populations suggest that K. pentacarpos might undergo a dynamics of population establishment and extinction in the Ricarda marshes. Because of fluctuating reproduction and the lack of a persistent seed bank, the conservation of standing adult populations appears to be a key factor to ensure the persistence of the species.