Dipeptidyl peptidase (DPP) IV in rat organs. Comparison of immunohistochemistry and activity histochemistry

Immunohistochemistry and activity histochemistry were used to study the localization of dipeptidyl peptidase (DPP) IV in rats. For immunohistochemistry, polyclonal as well as monoclonal anti-DPP IV antibodies were employed. The pattern of DPP IV immunoreactivity, determined with polyclonal anti-DPP IV antibody, corresponds to the histochemical pattern found for the enzymic activity of DPP IV. Immunoreactivity was present, in addition, in nerve cells, lateral membranes of certain surface epithelia, e.g., Fallopian tube, uterus and vesicular gland, in the luminal cytoplasm of e.g., vesicular gland epithelium, and in mucous cells of Brunner's gland. The monoclonal antibodies against DPP IV recognized four different epitopes (A-D) of the DPP IV molecule, and revealed that certain epitopes were not detectable by immunohistochemistry in some organs. Generally, the staining intensities for epitopes A, B, C and D decreased in that order. Usually, the monoclonal antibodies against epitopes A and B showed similar reaction patterns to those as obtained with the polyclonal antibody. Epitope D was recognized in the lumen of the duct system of exocrine glands and the intestine. Furthermore, high reactivity of this epitope was detected in goblet cells of the intestine, where no DPP IV activity was present.     

Effect of Xanthobacter,isolated and characterized from rice roots,on growth of wetland rice

With an autotrophic, N-free medium, Xanthobacter populations were isolated from the roots of wetland rice grown under field conditions. Xanthobacter populations ranged from 3.2×10⁴ to 5.1×10⁵ colony-forming units (cfu) g^-1 of root and averaged 47-fold higher on the root or rhizoplane than in the neighbouring nonrhizosphere. Characterization studies indicated dissimilarities in carbon utilization and motility among the isolated Xanthobacter strains and other recognized Xanthobacter species. Under gnotobiotic conditions, the population of one isolate, Xanthobacter sp. JW-KR1, increased from 10⁵ to 10⁷ cfu plant^-1 1 d after inoculation when a rice plant was present, but declined to numbers below the limit of detection (<10⁴ cfu assembly^-1) after 3 d in the absence of a plant. Scanning electron microscopy revealed Xanthobacter as pleomorphic forms on the rhizoplane. To assess the effect of Xanthobacter on plant growth, rice plants were grown under greenhouse conditions in plant assemblies containing sand and half-strength Hoagland's nutrient solution with and without nitrogen. Plants were either inoculated with 10⁵ cfu Xanthobacter g^-1 of sand or left uninoculated. After 40 d, plants without nitrogen showed no significant differences in top or root dry weight, plant height, root length, or number of tillers or leaves, whether the plants were inoculated or uninoculated. However, when nitrogen was added, inoculated plants had a significantly larger top dry weight (15%) and number of leaves (19%) than uninoculated plants. Under conditions of added and no added nitrogen, acetylene reduction assays showed Xanthobacter sp. JW-KR1 produced <0.1 (below detection limit) and 7 nmol C₂H₄ plant^-1 h^-1, respectively. Under the conditions studied, the results suggest that both Xanthobacter and wetland rice derive some benefits from their association.     

The Role of Microbes Associated with Chicken Litter in the Suppression of Meloidogyne arenaria

The role of microbes associated with chicken litter in the suppression of Meloidogyne arenaria in amended soil was investigated. Amended soil treatments were prepared, including combinations of sterile and nonsterile chicken litter and soil. Microbial biomass in different treatments was compared by measuring carbon dioxide evolution. There was less CO₂ evolved in sterile litter than in nonsterile litter treatments. Tomato seedlings cv. Rutgers were transplanted into soil mixtures and inoculated with 2,000 M. arenaria eggs. After 10 days, fewer second-stage juveniles (J2) had penetrated the roots in soils amended with nonsterile litter than sterile litter. The effects of sterile and nonsterile litter-amended soil solutions on M. arenaria eggs and J2 were observed over a period of 6 days. A lower percentage of eggs remained apparently healthy in nonsterile than in sterile-amended soil solutions over 6 days. Microbial degradation of the egg shells was apparent. Fewer J2 survived in sterile- and nonsterile-amended-soil solutions as compared to water controls.     

Growth of genetically engineered Pseudomonas aeruginosa and Pseudomonas putida in soil and rhizosphere.

The effect of the addition of a recombinant plasmid containing the pglA gene encoding an alpha-1,4-endopolygalacturonase from Pseudomonas solanacearum on the growth of Pseudomonas aeruginosa and Pseudomonas putida in soil and rhizosphere was determined. Despite a high level of polygalacturonase production by genetically engineered P. putida and P. aeruginosa, the results suggest that polygalacturonase production had little effect on the growth of these strains in soil or rhizosphere.     

Consequences of chlorophyll deficiency for leaf carotenoid composition in tobacco synthesizing glutamate 1-semialdehyde aminotransferase antisense RNA: dependency on developmental age and growth light

Transgenic tobacco (Nicotiana tabacum), with a reduced chlorophyll content of up to less than 10% of the wild-type level due to a different expression of antisense RNA coding for glutamate 1-semialdehyde aminotransferase, were used to study the relationship between chlorophyll accumulation and changes in carotenoid composition in developing and mature leaves grown either under low (30 mol photons m^-2 s^-1) or high light (300 mol photons m^-2 s^-1). Regardless of the extent to which chlorophyll synthesis was reduced, under low light the ratios of total chlorophyll to carotenoids remained constant. In contrast, under high light the content of carotenoids was elevated relative to chlorophyll and increased further with progressive inhibition in chlorophyll synthesis. The xanthophyll-cycle pigment pool was most strongly increased (up to 18-fold) upon suppression of chlorophyll synthesis. Concurrently to the higher pool sizes a higher extent of violaxanthin was converted into antheraxanthin and zeaxanthin and this was found to be correlated with a decrease in the quantum yield of photosystem II photochemistry. While lutein increased (up to 3-fold) with decreasing chlorophyll contents in high light transformants, neoxanthin remained rather constant in all plants analysed. Based on the present results, two different levels for the regulation of carotenoid synthesis are proposed depending on (I) the chlorophyll synthesizing capacity, and (ii) the photosynthetic light utilization efficiency. The first point suggests a co-regulation between carotenoid and chlorophyll synthesis; the second emphasizes the special role of carotenoids for protection against light stress.     

Ecosystem services provided by semi-natural and intensified grasslands: Synergies,trade-offs and linkages to plant traits and functional richness

Question

Semi-natural grasslands (SNG) are important for maintaining biodiversity and ecological processes in farmland. Current pasture-based livestock production mainly occurs on intensified grasslands (IG) that have been agronomically improved. Although it is documented that SNG and IG differ in terms of plant diversity, their ability to provide ecosystem services (ES) in farmland is less explored. Here, we review the role of SNG and IG in delivering ES, illustrate their trade-offs and synergies, and examine how ES can be assessed by using plant traits and functional richness.

Results

We found that SNG generate a wider range of ES than IG. Trade-offs exist between ES that appear more pronounced in IG between high biomass production and other ES. SNG are good in providing habitat for biodiversity, supporting pollination and cultural services. SNG also have a significantly wider range of plant functional traits and a higher functional richness, suggesting that the potential to supply ES in SNG is partly driven by higher number of species and their functional diversity.

Conclusion

Clearer trade-offs were found in IG compared with SNG, supported both by the literature and the functional richness. This suggests that functional knowledge could be a good complement to understand the mechanisms behind ES generation and could help with tailoring grassland management to sustain biodiversity, ecological functions and ES. Although both IG and SNG are likely needed for the long-term sustainability of food production, both could aim for a more balanced generation of ES, increasing biodiversity and functional redundancy at the landscape scale.     

Choline acetyltransferase activity of spinal cord cell cultures increased by co-culture with muscle and by muscle-conditioned medium

Activity of the enzyme choline acetyltransferase (CAT), which mediates the synthesis of the neurotransmitter, acetylcholine, was increased up to 20- fold in spinal cord (SC) cells grown in culture with muscle cells for 2 wk. This increase was directly related to the duration of co-culture as well as to the cell density of both the SC and muscle involved and was not affected by the presence of the acetylcholine receptor blocking agent, α-bungarotoxin. Glutamic acid decarboxylase (GAD) activity was often markedly decreased in SC-muscle cultures while the activities of acetylcholinesterase and several other enzymes were little changed. Increased CAT activity was also observed when SC cultures were maintained in medium which had been conditioned by muscle cells or by undifferentiated cells from embryonic muscle. Muscle-conditioned medium (CM) did not affect the activities of SC cell GAD or acetylcholinesterase. Dilution or concentration of the CM directly affected its ability to increase SC CAT activity , as did the duration and timing of exposure of the SC cells to the CM. The medium could be conditioned by muscle cells in the presence or absence of serum, and remained effective after dialysis or heating to 58 degrees C. Membrane filtration data were consistent with the conclusion that the active material(s) in CM had a molecular weight in excess of 50,000 daltons. We conclude that large molecular weight material that is released by muscle cells is capable of producing a specific increase in CAT activity of SC cells.     

Phytoremediation of pyrene in a Cecil soil under field conditions

We evaluated the effects of annual ryegrass (Lolium multiflorum Lam.) and phosphorus (P) availability on the dissipation of pyrene added at a concentration of approximately 600 mg kg-1 dry soil in the top 7.5 cm of a Cecil loamy sand (fine, kaolinitic, thermic Typic Kanhapludults) in a 10-month experiment under field conditions in Clemson, South Carolina. Plastic canopies were installed to prevent flooding of plots and raindrop dispersion of pyrene. Treatment factors were pyrene, vegetation, and available P levels. Each of the eight treatments had four replicates. The soil was adjusted to low and high P concentrations (an average of 41 and 66 kg extractable P ha-1, respectively). After a 175-d lag period for all treatments, the rate of pyrene removal followed first-order kinetics. The first-order rate constant was significantly higher in nonvegetated (0.098 d-1) than vegetated treatments (0.034 d-1). These data suggest that the presence of easily biodegradable organic matter from plant roots slowed the removal rate of pyrene. The levels of available P did not affect the rate of pyrene dissipation. Pyrene decreased below the detection limit of 6.25 mg kg-1 dry soil in all treatments after 301 d.     

Kinetic and mechanistic basis of the nonprocessive Kinesin-3 motor NcKin3

Kinesin-3 motors have been shown to transport cellular cargo along microtubules and to function according to mechanisms that differ from the conventional hand-over-hand mechanism. To find out whether the mechanisms described for Kif1A and CeUnc104 cover the full spectrum of Kinesin-3 motors, we characterize here NcKin3, a novel member of the Kinesin-3 family that localizes to mitochondria of ascomycetes. We show that NcKin3 does not move in a K-loop-dependent way as Kif1A or in a cluster-dependent way as CeUnc104. Its in vitro gliding velocity ranges between 0.30 and 0.64 mum/s and correlates positively with motor density. The processivity index (k(bi,ratio)) of approximately 3 reveals that not more than three ATP molecules are hydrolyzed per productive microtubule encounter. The NcKin3 duty ratio of 0.03 indicates that the motor spends only a minute fraction of the ATPase cycle attached to the filament. Unlike other Kinesin-3 family members, NcKin3 forms stable dimers, but only one subunit releases ADP in a microtubule-dependent fashion. Together, these data exclude a processive hand-over-hand mechanism of movement and suggest a power-stroke mechanism where nucleotide-dependent structural changes in a single motor domain lead to displacement of the motor along the filament. Thus, NcKin3 is the first plus end-directed kinesin motor that is dimeric but moves in a nonprocessive fashion to its destination.     

Maternal prenatal depressive symptoms predict infant NR3C1 1F and BDNF IV DNA methylation

Prenatal maternal psychological distress increases risk for adverse infant outcomes. However, the biological mechanisms underlying this association remain unclear. Prenatal stress can impact fetal epigenetic regulation that could underlie changes in infant stress responses. It has been suggested that maternal glucocorticoids may mediate this epigenetic effect. We examined this hypothesis by determining the impact of maternal cortisol and depressive symptoms during pregnancy on infant NR3C1 and BDNF DNA methylation. Fifty-seven pregnant women were recruited during the second or third trimester. Participants self-reported depressive symptoms and salivary cortisol samples were collected diurnally and in response to a stressor. Buccal swabs for DNA extraction and DNA methylation analysis were collected from each infant at 2 months of age, and mothers were assessed for postnatal depressive symptoms. Prenatal depressive symptoms significantly predicted increased NR3C1 1F DNA methylation in male infants (β = 2.147, P = 0.044). Prenatal depressive symptoms also significantly predicted decreased BDNF IV DNA methylation in both male and female infants (β = −3.244, P = 0.013). No measure of maternal cortisol during pregnancy predicted infant NR3C1 1F or BDNF promoter IV DNA methylation. Our findings highlight the susceptibility of males to changes in NR3C1 DNA methylation and present novel evidence for altered BDNF IV DNA methylation in response to maternal depression during pregnancy. The lack of association between maternal cortisol and infant DNA methylation suggests that effects of maternal depression may not be mediated directly by glucocorticoids. Future studies should consider other potential mediating mechanisms in the link between maternal mood and infant outcomes.