Adherens junctions and desmosomes are damaged by Entamoeba histolytica: Participation of EhCPADH complex and EhCP112 protease

Entamoeba histolytica trophozoites adhere to epithelium at the cell–cell contact and perturb tight junctions disturbing the transepithelial electrical resistance. Behind tight junctions are the adherens junctions (AJs) that reinforce them and the desmosomes (DSMs) that maintain the epithelium integrity. The damage produced to AJs and DMSs by this parasite is unknown. Here, we studied the effect of the trophozoites, the EhCPADH complex, and the EhCP112 recombinant enzyme (rEhCP112) on AJ and DSM proteins. We found that trophozoites degraded β‐cat, E‐cad, Dsp l/ll, and Dsg‐2 with the participation of EhCPADH and EhCP112. After contact of epithelial cells with trophozoites, immunofluorescence and transmission electron microscopy assays revealed EhCPADH and rEhCP112 at the intercellular space where they colocalised with β‐cat, E‐cad, Dsp l/ll, and Dsg‐2. Moreover, our results suggested that rEhCP112 could be internalised by caveolae and clathrin‐coated vesicles. Immunoprecipitation assays showed the interaction of EhCPADH with β‐cat and Dsp l/ll. Besides, in vivo assays demonstrated that rEhCP112 concentrates at the cellular borders of the mouse intestine degrading E‐cad and Dsp I/II. Our research gives the first clues on the trophozoite attack to AJs and DSMs and point out the role of the EhCPADH and EhCP112 in the multifactorial event of trophozoites virulence.     

Mapping of homoeologous chromosome exchanges influencing quantitative trait variation in Brassica napus

Genomic rearrangements arising during polyploidization are an important source of genetic and phenotypic variation in the recent allopolyploid crop Brassica napus. Exchanges among homoeologous chromosomes, due to interhomoeologue pairing, and deletions without compensating homoeologous duplications are observed in both natural B. napus and synthetic B. napus. Rearrangements of large or small chromosome segments induce gene copy number variation (CNV) and can potentially cause phenotypic changes. Unfortunately, complex genome restructuring is difficult to deal with in linkage mapping studies. Here, we demonstrate how high‐density genetic mapping with codominant, physically anchored SNP markers can detect segmental homoeologous exchanges (HE) as well as deletions and accurately link these to QTL. We validated rearrangements detected in genetic mapping data by whole‐genome resequencing of parental lines along with cytogenetic analysis using fluorescence in situ hybridization with bacterial artificial chromosome probes (BAC‐FISH) coupled with PCR using primers specific to the rearranged region. Using a well‐known QTL region influencing seed quality traits as an example, we confirmed that HE underlies the trait variation in a DH population involving a synthetic B. napus trait donor, and succeeded in narrowing the QTL to a small defined interval that enables delineation of key candidate genes.     

Relationship between vegetation structure and microenvironment in Fagus grandifolia subsp. mexicana forest relicts in Mexico

Aims Changes in the structure and composition of forests, whether caused by natural or anthropic events, alter the microenvironment, sometimes irreversibly. Since the local environment has a direct impact on basic ecological processes, this has become a key component of research. Mexican beech forests (Fagus grandifolia subsp. mexicana) in the Sierra Madre Oriental are restricted to sites with specific climate, soils and topography, making them an ideal natural system for ecological research. The objectives of this study were to identify the relationship between the microenvironment and the tree and shrub structure and composition of Mexican beech forests in the state of Hidalgo, and to compare the floristic similarity of these forests on the country scale using data from seven localities.Methods Specimens were collected for a period of one year at all localities in the state of Hidalgo where beech forests are located. At each locality, five 400 m 2 plots were established, and structural attributes (basal area, coverage, density and species richness) and six environmental variables were measured in the plots. The relationship between structure and microenvironment was estimated by simple correlation and canonic correspondence analysis (CCA). In addition, floristic similarity between different beech forest localities in the Sierra Madre Oriental was estimated by correspondence analysis (CA).Important findings Twenty tree species and eight shrub species were identified; at all localities studied F. grandifolia subsp. mexicana dominated the canopy. The multivariate analysis indicated that (i) in the four localities in the state of Hidalgo, all microenvironmental variables except pH are related to the variation observed in species composition and structure; (ii) the El Gosco locality had both tree and shrub species and microenvironmental factors different from those observed in the Fagus forests at the other localities in the study and (iii) the localities studied in order to draw country-scale comparisons could be divided into three groups by floristic similarity. The first group consisted of the Hidalgo localities, the second of the Veracruz localities, and the third, more different from the others, of the Tamaulipas locality. The results of this study provide the first reference for the relationship between the range of microenvironments and species structure in Mexican beech forests. Microenvironmental conditions in the larger beech forests could be used as a model for designing management and conservation programs for this plant association. Because of its particular ecological and historical characteristics, this association could serve as an example of biodiversity conservation in Mexico.     

Longitudinal study on growth in height in Polish boys and girls

Physical growth and development of children, determined by endogenous factors (genetic and paragenetic) is modified by numerous exogenic ones. This affects differentiation of ways of growth in individual children. In order to estimate the individual growth patterns and changes in growth rate as well as differences in growth of boys and girls, one must observe the process of growth of the same children and over a period of many years (longitudinal study). Such longitudinal study is the only way to provide information about the variability of growth and growth velocity at each age.     

Life table and biological characteristics of Macrolophus pygmaeus when feeding on Myzus persicae and Trialeurodes vaporariorum

The life table and biological characteristics of the predatory bug Macrolophus pygmaeus Rambur (Hemiptera: Miridae) were studied when the bugs were fed with Myzus persicae (Sulzer) (Homoptera: Aphididae) feeding on eggplant and with Trialeurodes vaporariorum Westwood (Homoptera: Aleyrodidae) feeding on tomato plants. The tests were done at five temperatures between 15 and 30 °C, using a L16:D8 photoperiod and 65 ± 5% r.h. Most eggs (range 85 to 90%) hatched at 15 and 20 °C. Incubation period was shortest at 27.5 °C (8.45 and 8.38 days on eggplant and tomato, respectively). Preoviposition was also shortest at 27.5 °C (5.10 and 4.75 days on eggplant and tomato, respectively) whereas fecundity was highest at 20 °C (213.90 and 228.25 eggs on eggplant and tomato, respectively). Maximum longevity of females was at 15 °C (122.40 and 129.35 days on eggplant and tomato, respectively). Mean generation time was longest at 15 °C on both host plants (122.75 and 124.64 days, respectively). The intrinsic rate of increase of M. pygmaeus was highest at 27.5 °C with similar values on eggplant (0.0981 day^–1) and tomato (0.1040 day^–1). Doubling time was shortest at 27.5 °C (7.06 and 6.67 days on eggplant and tomato, respectively) and, also, finite rate of increase was highest at 27.5 °C (1.1031 and 1.1096 on eggplant and tomato, respectively). The results show that the predator M. pygmaeus develops well on the aphid M. persicae or on the whitefly T. vaporariorum, both of which are important pests of vegetable crops. This predator is also well adapted to the temperatures that occur both in greenhouses and in the open field in the Mediterranean region. Compared to other natural enemies of whiteflies, such as Encarsia formosa Gahan (Hymenoptera: Aphelinidae), Macrolophus pygmaeus can increase at relatively low temperatures.     

Effects of infantile/prepubertal chronic estrogen treatment and chemical sympathectomy with guanethidine on developing cholinergic nerves of the rat uterus.

The innervation of the uterus is remarkable in that it exhibits physiological changes in response to altered levels in the circulating levels of sex hormones. Previous studies by our group showed that chronic administration of estrogen to rats during the infantile/prepubertal period provoked, at 28 days of age, an almost complete loss of norepinephrine-labeled sympathetic nerves, similar to that observed in late pregnancy. It is not known, however, whether early exposure to estrogen affects uterine cholinergic nerves. Similarly, it is not known to what extent development and estrogen-induced responses in the uterine cholinergic innervation are affected by the absence of sympathetic nerves. To address this question, in this study we analyzed the effects of infantile/prepubertal chronic estrogen treatment, chronic chemical sympathectomy with guanethidine, and combined sympathectomy and chronic estrogen treatment on developing cholinergic nerves of the rat uterus. Cholinergic nerves were visualized using a combination of acetylcholinesterase histochemistry and the immunohistochemical demonstration of the vesicular acetylcholine transporter (VAChT). After chronic estrogen treatment, a well-developed plexus of cholinergic nerves was observed in the uterus. Quantitative studies showed that chronic exposure to estrogen induced contrasting responses in uterine cholinergic nerves, increasing the density of large and medium-sized nerve bundles and reducing the intercept density of fine fibers providing myometrial and perivascular innervation. Estrogen-induced changes in the uterine cholinergic innervation did not appear to result from the absence/impairment of sympathetic nerves, because sympathectomy did not mimic the effects produced by estrogen. Estrogen-induced responses in parasympathetic nerves are discussed, considering the direct effects of estrogen on neurons and on changes in neuron-target interactions.     

Somatic Embryogenesis and Morphogenetic Potential of Spring Barley (Hordeum vulgare L.) in the System of Technological Genetic Transformation

We studied somatic embryogenesis and morphogenetic potential in young embryos of 17 spring barley cultivars. A considerable effect of the genotype as well as of certain biologically active compounds in the growth medium on the frequency of embryogenic callus formation and regenerative capacity was observed. The Murashige and Skoog medium complemented with myoinositol and casein hydrolysate is suitable for embryogenic callus induction. The regeneration medium should be complemented by hormones, 2.5 mg/l auxin 2,4-D and 0.1–0.5 mg/l cytokinin 6-BAP, to increase the frequency of somatic embryo maturation. The presence of cefotaxime and high level of copper are desirable to increase the efficiency of callogenesis, regeneration, and callus quality. Three cultivars with economically valuable characters and high morphogenetic potential were recommended for development of efficient technology for barley transformation.     

Effect of dissolved oxygen concentration on differentiation of somatic embryos of Coffea arabica cv. Catimor 9722

The effect of two different dissolved oxygen (DO) concentrations (50 and 80%) on differentiation of somatic embryos (SE) from cell suspensions of coffee (Coffea arabica cv. Catimor 9722) was analyzed. Two bioreactors CMF-100 (CHEMAP AG) designed for the culture of cells, with 2-l glass vessels and a maximum work volume of 1.8 l were used. Each one was equipped with a gas blending unit (air, O₂, N₂, CO₂) for the control of DO concentration. The inoculation density of embryogenic cells was 1.0 gram of fresh weight per liter (g FW l^–1). The number of somatic embryos was greater (71 072 SE l^–1) with 80% DO, but the major proportion were globular and heart shaped SE (66 399 SE l^–1) and only 6.6% with regard to total was torpedo shaped SE. However, the 50% DO produced the higher number in the torpedo shaped SE (7389 SE l^–1) what represented 20.0% with regard to total. Thus, higher concentrations of DO induced globular and heart shaped SE differentiation, but for production of torpedo shaped SE lower concentrations DO are needed. The somatic embryos obtained in the bioreactor with 50% DO showed similar behavior to the somatic embryos obtained in the rotary shaker. After 8 weeks of culture, 49.2% germination was obtained, which allowed a total of 1725 plantlet to be transferred to conditions ex vitro. After 6 months of culture, 89.2% of conversion was achieved and 1539 plants obtained were transferred to field conditions.     

Associations between canopy and understory species increase along a rainshadow gradient in the Alps: habitat heterogeneity or facilitation?

Spatialassociations among overstory and understory species tend to increase ongradients from wet to dry climates. This shift in the strength of spatialassociations has usually been attributed to shared abiotic requirements betweencanopy species and understory assemblages within communities and/or to anincrease in habitat heterogeneity in dry climates and therefore higher betadiversity. On another hand, more important positive effects of tree canopies onunderstory species in drier climates may also explain stronger associations andhigher beta diversity. We examined these three hypotheses along a strongrainshadow gradient that occurs from the wet external Alps to the dry innerAlpsby analyzing with correspondence analysis and canonical correspondence analysisthe species composition of 290 relevés of forests dominated to differentdegrees by Abies alba and Piceaabies.We found important differences in climatic requirements forAbies and Picea, withAbies occurring in warmer and drier habitats thanPicea. The understory species associated with these twospecies showed similar correlations with temperature but not with moisture,withunderstory species of Picea-communities having strongerxeric affinities than understory species ofAbies-communities. We found no significant associationsbetween canopy species and understory composition in the external Alps despitethe fact that Abies and Piceaoccurredin substantially different environments. In contrast,Abiesand Picea occurred in more similar environments in theinner Alps, but the understory assemblages associated with eitherAbies or Picea were significantlydifferent. This increase in canopy-understory associations was in partdetermined by strong differences in moisture between southern and northernaspects in the inner Alps, which affected both canopy and understory speciesdistributions. However, differences between the canopy effects ofPicea and Abies also appeared tocontribute to stronger associations between canopy and understory species, andconsequently to increase beta diversity. This pattern only occurred on southernaspects of the inner Alps but was highly significant. Our results suggest thatspecies distributions may be continuous on the wet ends of moisture gradientsbut discrete on dry ends. Relatively discrete communities at stressful ends ofgradients appear to develop as a result of both habitat differentiation and thepositive effects of overstory species.