Phytochemical Analysis and in vitro Free‐Radical‐Scavenging Activities of the Essential Oils from Leaf and Fruit of Melaleuca leucadendra L.

The phytochemical profile of Melaleuca leucadendra L. leaf and fruit oils from Cuba was investigated by GC and GC/MS. Forty‐one and sixty‐four volatile compounds were identified and quantified, accounting for 99.2 and 99.5% of the leaf‐oil and fruit‐oil total composition, respectively. The main components were 1,8‐cineol (43.0%), viridiflorol (24.2%), α‐terpineol (7.0%), α‐pinene (5.3%), and limonene (4.8%) in the leaf oil, and viridiflorol (47.6%), globulol (5.8%), guaiol (5.3%), and α‐pinene (4.5%) in the fruit oil. The antioxidant capacity of these essential oils was determined by three different in vitro assays (2,2‐diphenyl‐1‐picrylhydrazyl (DPPH) radical, thiobarbituric acid reactive species (TBARS), and 2,2′‐Azinobis(3‐ethylbenzothiazoline‐6‐sulfonic acid) (ABTS) radical cation), and significant activities were evidenced for all of them.     

Understorey plant species richness and composition in metropolitan forest archipelagos: effects of forest size, adjacent land use and distance to the edge

Aim To address the relative role of adjacent land use, distance to forest edge, forest size and their interactions on understorey plant species richness and composition in perimetropolitan forests. Location The metropolitan area of Barcelona, north‐eastern Spain. Methods Twenty sampling sites were distributed in two forest size‐categories: small forest patches (8–90 ha) and large forest areas (> 18,000 ha). For each forest‐size category, five sites were placed adjacent to crops and five sites adjacent to urban areas. Vascular plant species were recorded and human frequentation was scored visually in 210 10 × 10 m plots placed at 10, 50 and 100 m from the forest edge, and additionally at 500 m in large forest areas. Plant species were grouped according to their ecology and rarity categories. A nonmetric multidimensional scaling (NMS) ordination was carried out to detect patterns of variation in species assemblage, and to explore the relationships between these patterns and the richness of the species groups and the studied factors. Factorial anovas were used to test the significance of the studied factors on the richness of species groups. Relationships between human frequentation and the studied variables were assessed through contingency tables. Results Forest‐size category was the main factor affecting synanthropic species (i.e. those thriving in man‐made or man‐disturbed habitats). Synanthropic species richness decreased with increasing distance from the forest edge and, when forests were adjacent to crops, it was higher in small forest patches than in large forest areas. Richness of rare forest species was lower in small forest patches than in large forest areas when forests were adjacent to urban areas. Richness of common forest species and of all forest species together were higher close to the forest edge than far from it when forests were adjacent to urban areas. Forests adjacent to urban areas were more likely to experience high human frequentation, particularly in those plots nearest to the forest edge. Main conclusions Forest‐size category and adjacent land use were the most important factors determining species richness and composition. The preservation of large forests adjacent to crops in peri‐urban areas is recommended, because they are less frequented by humans, are better buffered against the percolation of nonforest species and could favour the persistence of rare forest species.     

Risk markers in subjects with high loads of Ascaris lumbricoides in a rural community of the Cojedes State , Venezuela

A total of 337 individuals from a rural community of Venezuela (Mapurite, Estado Cojedes) was studied. Stoll faecal examination was carried out and the total number of A. lumbricoides was recovered after the treatment of the individuals sampled with Pyrantel pamoate at a dose of 10 mg/kg body weight in children and 15 mg/kg in adults. A relationship between the high worm burden condition (individuals excreting more than 10,000 epg of A. lumbricoides) and the age of the hosts was found. Resulting with the major values of relative risk index (Rr) the 5-9 years old class (Rr = 3.2 t = 3.4). This relationship was not found between high worm burden condition (HWB) and the sex of the hosts. The following individuals have the the highest possibilities to become HWB: with blood group A (Rr = 3.05 t = 4.08), with haemoglobin type HbA HbS (Rr = 1.86 t = 2.13), individuals with the combinations A HbA HbF (Rr = 3.46 t = 2.31) and A HbA HbS (Rr = 3.11 t = 2.78). It was estimated that the selective treatment of the total HWB detected (72) with an effective product, determined a reduction of 95.4% of the environmental contamination with de eggs and 72.3% of the worm burden.     

Uridine uptake by isolated intestinal epithelial cells of guinea pig

Uptake of uridine was studied in isolated intestinal epithelial cells of guinea pig. Uptake was not severely influenced by metabolism. Free uridine was accumulated within cells 13-fold. Uptake was saturable with an apparent K_m value of 46 μM and a V_max of 0.9 nmol/mg protein per min. Uracil inhibited uptake only slightly; adenosine, guanosine and cytosine inhibited strongly. Antimycin A and ouabain inhibited almost 90%. If the extracellular Na⁺ concentration was decreased to 5 mM, the rate of uptake decreased 6.5-fold. The stimulatory effect of Na⁺ was related to the transmembraneous Na⁺-gradient. Cells from jejunum transported about 30% faster than cells from ileum. In conclusion, isolated enterocytes of guinea pig posses an active transport system for uridine.     

Cdc42 GTPase-activating proteins (GAPs) regulate generational inheritance of cell polarity and cell shape in fission yeast

The highly conserved small GTPase Cdc42 regulates polarized cell growth and morphogenesis from yeast to humans. We previously reported that Cdc42 activation exhibits oscillatory dynamics at cell tips of Schizosaccharomyces pombe cells. Mathematical modeling suggests that this dynamic behavior enables a variety of symmetric and asymmetric Cdc42 activation distributions to coexist in cell populations. For individual wild-type cells, however, Cdc42 distribution is initially asymmetrical and becomes more symmetrical as cell volume increases, enabling bipolar growth activation. To explore whether different patterns of Cdc42 activation are possible in vivo, we examined S. pombe rga4∆ mutant cells, lacking the Cdc42 GTPase-activating protein (GAP) Rga4. We found that monopolar rga4∆ mother cells divide asymmetrically leading to the emergence of both symmetric and asymmetric Cdc42 distributions in rga4∆ daughter cells. Motivated by different hypotheses that can mathematically reproduce the unequal fate of daughter cells, we used genetic screening to identify mutants that alter the rga4∆ phenotype. We found that the unequal distribution of active Cdc42 GTPase is consistent with an unequal inheritance of another Cdc42 GAP, Rga6, in the two daughter cells. Our findings highlight the crucial role of Cdc42 GAP localization in maintaining consistent Cdc42 activation and growth patterns across generations.     

The sarcosine effect on protein stability: A case of nonadditivity?

We have used differential scanning calorimetry to determine the effect of low concentrations (C = 0-2 M) of the osmolyte sarcosine on the Gibbs energy changes (deltaG) for the unfolding of hen-egg-white lysozyme, ribonuclease A, and ubiquitin, under the same buffer and pH conditions. We have also computed this effect on the basis of the additivity assumption and using published values of the transfer Gibbs energies for the amino acid side chains and the peptide backbone unit. The values thus predicted for the slope delta deltaG/deltaC agree with the experimental ones, but only if the unfolded state is assumed to be compact (that is, if the accessibility to solvent of the unfolded state is modeled using segments excised from native structures). The additivity-based calculations predict similar delta deltaG/deltaC values for the three proteins studied. We point out that, to the extent that this approximate constancy of delta deltaG/deltaC holds, osmolyte-induced increases in denaturation temperature will be larger for proteins with low unfolding enthalpy (small proteins that bury a large proportion of apolar surface). The experimental results reported here are consistent with this hypothesis.     

High frequency of homozygosity of the HLA region in melanoma cell lines reveals a pattern compatible with extensive loss of heterozygosity

Malignant transformation of cells is frequently associated with abnormalities in human leukocyte antigen (HLA) expression. MHC class I loss or down-regulation in cancer cells is a major immune escape route used by a large variety of human tumours to evade antitumour immune responses mediated by cytotoxic T lymphocytes. The goal of our study was to explore HLA genotyping and phenotyping in a variety of melanoma tumour cell lines. A total of 91 melanoma cell lines were characterised for HLA class I and II genotype. In addition, 61 out of the 91 cell lines were also analysed for HLA class I and II cell surface molecule expression by flow cytometry. Unexpectedly, we found that 19.7% of the melanoma cell lines were homozygous for HLA class I genotypes, sometimes associated with HLA class II homozygosity (8.79%) and sometimes not (10.98%). The frequency of homozygosity was significantly higher compared with the control groups (1.6%). To identify the reasons underlying the high frequency of HLA homozygosity we searched for genomic deletions using eight pairs of highly polymorphic microsatellite markers covering the entire extended HLA complex on the short arm of chromosome 6. Our results were compatible with hemizygous deletions and suggest that loss of heterozygosity on chromosome arm 6p is a common feature in melanoma cell lines. In fact, although autologous normal DNA from the patients was not available and could not be tested, the retention in some cases of heterozygosity for a number of microsatellite markers would indicate a hemizygous deletion. In the rest of the cases, markers at 6p and 6q showed a single allele pattern indicating the probable loss of part or the whole of chromosome 6. These results led us to conclude that loss of heterozygosity in chromosome 6 is nonrandom and is possibly an immunologically relevant event in human malignant melanoma. Other well-established altered HLA class I phenotypes were also detected by flow cytometry that correspond to HLA class I total loss and HLA-ABC and/or specific HLA-B locus down-regulation.