Gastrointestinal absorption and metabolism of apple polyphenols ex vivo by the pig intestinal mucosa in the Ussing chamber

Polyphenols contained in food have various positive effects on human health. The absorption and metabolism of polyphenols in the intestinal tract needs to be studied to estimate these effects. The Ussing chamber technique was used to investigate the transport behavior of apple polyphenols through pig small intestinal mucosa, which served as a model for human gastrointestinal mucosa. The identities and concentrations of polyphenols and their metabolites in the half-chambers (luminal and basolateral) within an incubation period of 4 h were determined by HPLC–MS/MS and HPLC–DAD (DAD = diode-array detection). Flux values were also measured. It was found that 5-caffeoylquinic acid and caffeic acid were absorbed and translocated to the basolateral side (1.9 and 3.7%, respectively), but other compounds, including glycosides of phloretin and quercetin, were observed without translocation. A Ussing chamber utilizing pig small intestinal mucosa is a suitable model for assessing the effect of apple polyphenols on mucosal integrity and nutrition absorption across porcine mucosa.     

Air humidity as key determinant of morphogenesis and productivity of the rare temperate woodland fern Polystichum braunii

(1) Most ferns are restricted to moist and shady habitats, but it is not known whether soil moisture or atmospheric water status are decisive limiting factors, or if both are equally important. (2) Using the rare temperate woodland fern Polystichum braunii, we conducted a three‐factorial climate chamber experiment (soil moisture (SM) × air humidity (RH) × air temperature (T)) to test the hypotheses that: (i) atmospheric water status (RH) exerts a similarly large influence on the fern's biology as soil moisture, and (ii) both a reduction in RH and an increase in air temperature reduce vigour and growth. (3) Nine of 11 morphological, physiological and growth‐related traits were significantly influenced by an increase in RH from 65% to 95%, leading to higher leaf conductance, increased above‐ and belowground productivity, higher fertility, more epidermal trichomes and fewer leaf deformities under high air humidity. In contrast, soil moisture variation (from 66% to 70% in the moist to ca. 42% in the dry treatment) influenced only one trait (specific leaf area), and temperature variation (15 °C versus 19 °C during daytime) only three traits (leaf conductance, root/shoot ratio, specific leaf area); RH was the only factor affecting productivity. (4) This study is the first experimental proof for a soil moisture‐independent air humidity effect on the growth of terrestrial woodland ferns. P. braunii appears to be an air humidity hygrophyte that, whithin the range of realistic environmental conditions set in this study, suffers more from a reduction in RH than in soil moisture. A climate warming‐related increase in summer temperatures, however, seems not to directly threaten this endangered species.     

Autism‐related behaviors in the cyclooxygenase‐2‐deficient mouse model

Prostaglandin E2 (PGE2) is an endogenous lipid molecule involved in normal brain development. Cyclooxygenase‐2 (COX2) is the main regulator of PGE2 synthesis. Emerging clinical and molecular research provides compelling evidence that abnormal COX2/PGE2 signaling is associated with autism spectrum disorder (ASD). We previously found that COX2 knockout mice had dysregulated expression of many ASD genes belonging to important biological pathways for neurodevelopment. The present study is the first to show the connection between irregular COX2/PGE2 signaling and autism‐related behaviors in male and female COX2‐deficient knockin, (COX)‐2^?, mice at young (4‐6 weeks) or adult (8‐11 weeks) ages. Autism‐related behaviors were prominent in male (COX)‐2^? mice for most behavioral tests. In the open field test, (COX)‐2^? mice traveled more than controls and adult male (COX)‐2^? mice spent less time in the center indicating elevated hyperactive and anxiety‐linked behaviors. (COX)‐2^? mice also buried more marbles, with males burying more than females, suggesting increased anxiety and repetitive behaviors. Young male (COX)‐2^? mice fell more frequently in the inverted screen test revealing motor deficits. The three‐chamber sociability test found that adult female (COX)‐2^? mice spent less time in the novel mouse chamber indicative of social abnormalities. In addition, male (COX)‐2^? mice showed altered expression of several autism‐linked genes: Wnt2, Glo1, Grm5 and Mmp9. Overall, our findings offer new insight into the involvement of disrupted COX2/PGE2 signaling in ASD pathology with age‐related differences and greater impact on males. We propose that (COX)‐2^? mice might serve as a novel model system to study specific types of autism.     

T lymphocytes migrate upstream after completing the leukocyte adhesion cascade

The leukocyte adhesion cascade is of critical importance for both the maintenance of immune homeostasis and the ability of immune cells to perform effector functions. Here, we present data showing CD4⁺ T cells migrate upstream (against the direction of flow) after completing the leukocyte adhesion cascade on surfaces displaying either ICAM-1 or ICAM-1 and VCAM-1, but migrate downstream on surfaces displaying only VCAM-1. Cells completing the cascade on HUVECs initially migrate upstream before reverting to more random migration, partly caused by transmigration of cells migrating against the flow. Furthermore, cells migrating upstream transmigrate faster than cells migrating downstream. On HUVECs, blocking interactions between LFA-1 and ICAM-1 resulted in downstream migration and slower transmigration. These results further suggest a possible physiological role for upstream migration in vivo.     

Multichannel automated chamber system for continuous monitoring of CO2 exchange between the agro-ecosystem or soil and the atmosphere

A multichannel automated chamber system was developed for continuous monitoring of CO₂ exchange at multiple points between agro-ecosystem or soil and atmosphere. This system consisted of an automated chamber subsystem with a CO₂ concentration analyzer and a data logging subsystem. Both subsystems were under the control of a programmable logic controller (PLC). The automated chamber subsystem contained 18 chambers (50 cm × 50 cm × 50 cm) and a compressor. The chamber lids were closed and can be automatically opened. During measurement, one of the 18 chambers was kept closed for three minutes for measuring and the other chambers were kept open to maintain the natural soil conditions to the maximum extent. Environmental variables were simultaneously measured using sensors and recorded by the data logger. The reliability of the multichannel automated chamber system was tested and the results showed that the turbulence of the fans had no significant effect on the CO₂ exchange. The changes in the air and the temperature of soil and soil moisture inside the chambers, caused by the enclosure of the chambers, were not significant. The net ecosystem CO₂ exchange for the wheat ecosystem was ?2.35 μmol·m^?2·s^>?1 and the soil respiration was 3.87 μmol·m^?2·s^>?1 in the wheat field, and 6.61 μmol·m^?2·s^>?1 in the apple orchard.     

Effects of Elevated Carbon Dioxide on the Growth and Foliar Chemistry of Transgenic Bt Cotton

A field study was carried out to quantify plant growth and the foliar chemistry of transgenic Bacillus thuringiensis （Bt） cotton （cv. GK-12） exposed to ambient CO2 and elevated （double-ambient） CO2 for different lengths of time （1, 2 and 3 months） in 2004 and 2005. The results indicated that CO2 levels significantly affected plant height, leaf area per plant and leaf chemistry of transgenic Bt cotton. Significantly, higher plant height and leaf area per plant were observed after cotton plants that were grown in elevated CO2 were compared with plants grown in ambient CO2 for 1, 2 and 3 months in the investigation. Simultaneously, significant interaction between CO2 level x investigating year was observed in leaf area per plant. Moreover, foliar total amino acids were increased by 14%, 13%, 11% and 12%, 14%, 10% in transgenic Bt cotton after exposed to elevated CO2 for 1, 2 or 3 months compared with ambient CO2 in 2004 and 2005, respectively. Condensed tannin occurrence increased by 17%, 11%, 9% in 2004 and 12%, 11%, 9% in 2005 in transgenic Bt cotton after being exposed to elevated CO2 for 1, 2 or 3 months compared with ambient CO2 for the same time. However, Bt toxin decreased by 3.0%, 2.9%, 3.1% and 2.4%, 2.5%, 2.9% in transgenic Bt cotton after exposed to elevated CO2 for 1, 2 or 3months compared with ambient CO2 for same time in 2004 and 2005, respectively. Furthermore, there was prominent interaction on the foliar total amino acids between the CO2 level and the time of cotton plant being exposed to elevated CO2. It is presumed that elevated CO2 can alter the plant growth and hence ultimately the phenotype allocation to foliar chemistical components of transgenic Bt cotton, which may in turn, affect the plant-herbivore interactions.     

DNA methylation as a possible mechanism affecting ability of natural populations to adapt to changing climate

Environmentally induced epigenetic variation has been recently recognized as a possible mechanism allowing plants to rapidly adapt to novel conditions. Despite increasing evidence on the topic, little is known on how epigenetic variation affects responses of natural populations to changing climate. We studied the effects of experimental demethylation (DNA methylation is an important mediator of heritable control of gene expression) on performance of a clonal grass, Festuca rubra, coming from localities with contrasting temperature and moisture regimes. We compared performance of demethylated and control plants from different populations under two contrasting climatic scenarios and explored whether the response to demethylation depended on genetic relatedness of the plants. Demethylation significantly affected plant performance. Its effects interacted with population of origin and partly with conditions of cultivation. The effects of demethylation also varied between distinct genotypes with more closely related genotypes showing more similar response to demethylation. For belowground biomass, demethylated plants showed signs of adaptation to drought that were not apparent in plants that were naturally methylated. The results suggest that DNA methylation may modify the response of this species to moisture. DNA methylation may thus affect the ability of clonal plants to adapt to novel climatic conditions. Whether this variation in DNA methylation may also occur under natural conditions, however, remains to be explored. Despite the significant interactions between population of origin and demethylation, our data do not provide clear evidence that DNA methylation enabled adaptation to different environments. In fact, we obtained stronger evidence of local adaptation in demethylated than in naturally‐methylated plants. As changes in DNA methylation may be quite dynamic, it is thus possible that epigenetic variation can mask plant adaptations to conditions of their origin due to pre‐cultivation of the plants under standardized conditions. This possibility should be considered in future experiments exploring plant adaptations.     

黄柄钙皮菌的生活史

利用基物培养、燕麦-琼脂培养技术及扫描电镜技术研究了黄柄钙皮菌的个体发育过程,在燕麦琼脂培养基上完成了从孢子到孢子的生活史。结果表明,生活史包括单核的黏变形体或游动胞、多核的营养体原质团以及孢子形成阶段。孢子球形,表面具疣突。孢子萌发为裂式,释放1黏变形体。黏变形体行变形运动,在有水的条件下,可转变为游动胞并游动。可观察到1长具极性的鞭毛。合子形成原质团。成熟原质团棕色。原质团类型为显型,具有扇形网络状菌脉。琼脂培养基上获得的黄柄钙皮菌孢子与野生型相似,并具有可育性。     

Cell separation mediated by differential rolling adhesion

Recently, we showed a correlation between the maturity of hematopoietic stem and progenitor cells during development and rolling efficiency on selectins. These findings motivated us to explore a novel separation that exploits differences in selectin-mediated rolling adhesion between populations of cells. We extend the use of a previously developed cell-free system to study the separation of populations of sialyl Lewis x (sLe(x))-coated microspheres designed to roll with different average velocities on L-selectin chimeric substrates under well-defined flow. Results show that a separation that exploits differences in average rolling velocities between cell or microsphere populations is attainable. Excellent recovery and purity values for the slower rolling, or more desirable, populations are obtained and can be estimated from rolling velocity measurements. We also assess the feasibility of a selectin-mediated separation of adult bone marrow cell populations using previously obtained rolling velocity and rolling flux data for CD34+ and CD34- adult bone marrow cells on L-selectin substrates. We believe that a cell separation mediated by differential rolling adhesion can be used to enrich populations of hematopoietic stem and progenitor cells from an adult bone marrow cell preparation and that this method possesses several major advantages over existing antibody-mediated cell-affinity chromatography technologies.     

Enumerating phytoplankton with an upright compound microscope using a modified settling chamber

A modified settling chamber is described which permits the use of an upright compound microscope for phytoplankton enumerations. The chamber is composed of a 75 × 51 mm rectangular or 70 mm round-glass microslide base with a 130 m thick piece of sheet styrene attached to the upper surface. A circular cell is cut into the styrene material making a 26 mm diameter chamber which is approximately 130–150 m deep. Settling procedures follow Utermöhl's technique, with the use of a 0.13–0.15 mm thick coverslip (50 × 45 mm) to cover the chamber. The overall thickness of the settling slide is 1.13 mm which does not impact on the optical requirements for most objectives, including oil immersion objectives. The chamber encloses a total volume of 69–80 mm³. No statistical differences are observed in cell, filament or colony counts between the new and traditional chambers. Furthermore, count comparisons at different cell concentrations using the new chambers give consistent results. Thus, the resolution and availability of an upright microscope makes the use of the new settling chamber an attractive method for phytoplankton counting, especially in teaching situations.