Tissue tropism of Helicobacter pullorum in specific pathogen-free chickens determined by culture and nucleic acid detection

Three-day-old specific pathogen-free chickens (n = 24) located in isolators were inoculated orally with Helicobacter pullorum. One group (n = 12) was infected with a H. pullorum field isolate from human origin, another one (n = 12) with the American Type Culture Collection H. pullorum reference isolate 51801 originating from chickens. Both isolates were positive for cytolethal distending toxin, investigated using a polymerase chain reaction (PCR). A third group (n = 4) was kept as a negative control. Starting on day 7 of life, birds from each group were euthanatized at different time points up to 35 days. Various organ samples were taken aseptically and processed by culture and a H. pullorum-specific PCR. In the group infected with the human isolate the nucleic acid of H. pullorum was detected in the caecal tonsils and caeca of 12 and 11 birds, respectively. Live bacteria were cultivated from the caecal tonsils and caeca of five birds 24 and 31 days postinfection. Live bacteria were also isolated from the heart of one bird, whereas PCR had to be used to detect the nucleic acid of H. pullorum in the gallbladder of four birds. No live bacteria were reisolated at any time from birds infected with the avian isolate, but bacterial nucleic acid was detected in the caeca of five birds and in the gallbladder of one. In both groups neither live H. pullorum nor its nucleic acid were detected in the liver, spleen, and duodenum. Compared to the avian H. pullorum isolate the human isolate proved to be more invasive. No obvious clinical symptoms or disease was seen in the chickens during the entire experiment. The reisolation of live bacteria at the end of the experiments indicates that H. pullorum could enter the food chain even after early infection in birds. Furthermore, PCR was demonstrated to be helpful in tracing these fastidious bacteria.     

Quantification of jasmonic acid by SPME in tomato plants stressed by ozone

Jasmonates are signalling molecules induced in plants as a response to various biotic and/or abiotic stresses. As ozone is known to activate defense responses in plants, we have monitored the concentration of jasmonic acid in tomato leaves during and after an acute exposure to this abiotic elicitor. In this experiment, we observed that the maximum induction of jasmonic acid in O3-fumigated plants occurred 9 h after the end of treatment and the concentration of jasmonic acid in stressed plants increased 13-fold. However, the level of endogenous methyl-jasmonate was constant during the observed period. The extraction and quantification of jasmonic acid as its methyl ester was performed by headspace-solid-phase microextraction (or HS-SPME) in combination with GC-FID and GC-MS. The sensitivity (LOD = 2 ng/g) of this method permitted the detection and quantification of jasmonic acid present in plant tissues at very low concentrations.     

Adhesion of Escherichia coli K88ab fimbriae to porcine enterocytes: effect on enzymatic activities of the brush border, and cyclic AMP and GMP levels

We have studied the cellular alterations, after in vitro adherence, of purified K88ab fimbriae to membranes of porcine enterocytes. Effects on enzymatic activities as disaccharydases and alkaline phosphatase show low changes. While cAMP levels were decreased (44%), guanylyl cyclase was increased (up to 200%), and levels of cGMP were in consequence significantly affected. This study support the role of cyclic GMP as intracellular mediator for adherence, and suggest their implication in disease, affecting a membrane-mediated mechanisms for guanylate cyclase activation, that is unique in the intestine.     

Mouse epidermal development: effects of retinoic acid exposure in utero

Epidermal morphogenesis was studied in vivo following prenatal exposure to retinoic acid (RA). In pregnant mice, a single oral dose of RA on day 11.5 of gestation failed to induce histological changes in fetal epidermal development except in epidermal thickness. Epidermal thickness increased from 16.5 days post-coitum (dpc) onwards, and temporal and spatial epidermal modifications in keratins K5 and K14 related to proliferative activity of keratinocytes were observed. An RA effect on cell proliferation was supported by a statistically significant increase in the number of epidermal S-phase cells, containing BrdU-incorporated DNA in RA-exposed mice compared with nonexposed animals. The prolonged in utero action of RA on epidermal proliferative activity in fetuses and newborns suggests a long-term RA effect that may play a role on the development and evolution of diseases in adult skin.     

Influence of micro-oxygenation treatment before oak aging on phenolic compounds composition, astringency, and color of red wine

Micro-oxygenation is usually applied to red wines as a cheaper alternative to oak aging. It has been suggested, however, that micro-oxygenation can also be used to complement oak aging in order to improve the quality of very astringent and herbaceous red wines. In this paper we study how applying the micro-oxygenation technique before oak aging affects the composition and quality of astringent red wines. When this technique is applied prior to oak aging, the wines have a slightly less intense red color and significantly higher levels of combined and free anthocyanins and ethyl-bridged anthocyanin-flavanol pigments. On the other hand, no differences in other newly formed pigments are found. Applying micro-oxygenation before oak aging does not affect the total proanthocyanidin concentration, but it produces wines with a slightly (though significantly) higher mean degree of proanthocyanidin polymerization and a drastically lower astringency. These wines also present a clearer impact of wood aromas.     

BKCa-Cav channel complexes mediate rapid and localized Ca2+-activated K+ signaling

Large-conductance calcium- and voltage-activated potassium channels (BKCa) are dually activated by membrane depolarization and elevation of cytosolic calcium ions (Ca2+). Under normal cellular conditions, BKCa channel activation requires Ca2+ concentrations that typically occur in close proximity to Ca2+ sources. We show that BKCa channels affinity-purified from rat brain are assembled into macromolecular complexes with the voltage-gated calcium channels Cav1.2 (L-type), Cav2.1 (P/Q-type), and Cav2.2 (N-type). Heterologously expressed BKCa-Cav complexes reconstitute a functional "Ca2+ nanodomain" where Ca2+ influx through the Cav channel activates BKCa in the physiological voltage range with submillisecond kinetics. Complex formation with distinct Cav channels enables BKCa-mediated membrane hyperpolarization that controls neuronal firing pattern and release of hormones and transmitters in the central nervous system.     

Genotype × environment interactions in populations possessing <Emphasis Type="Italic">Ga1</Emphasis>-<Emphasis Type="Italic">s</Emphasis> and <Emphasis Type="Italic">ga1</Emphasis> alleles for cross incompatibility in maize

Use of cross incompatibility in corn (Zea mays L.) by the Ga1-s allele may reduce cross-fertilization in specialty and conventional organic corn with pollen from genetically-modified (GM) corn. For effective use, information about environment, and genotype × environment effects on cross-fertilization by ga1 as well as heritability of cross incompatibility in maize is necessary. Our objective was to obtain this information. Four population pairs differing in their genotype at ga1 were evaluated for cross incompatibility with ga1 pollen in different environments. Populations were derived by crossing the recurrent parents B116, PHG35, ARZM16035:S19, and (CHZM05015:Mo17)Mo17 to Ga1-s donor parent Mo508W/Mo506W. Two replicates of each treatment were grown in the center of 952 m² fields planted with purple corn as an adventitious source of ga1/ga1 pollen. Open pollination was allowed and the amount of cross-fertilization estimated by averaging the percentage of purple seeds. Environment and genotype × environment effects were not significant. Contrasts to evaluate differences in cross-fertilization between Ga1-s and ga1 populations revealed that mean percentages of cross-fertilization in Ga1-s populations of B116, ARZM16035:S19, and (CHZM05015:Mo17)Mo17 were significantly lower than in ga1 populations. The estimated broad-sense heritability on an entry-mean basis for cross incompatibility was 0.81. Results suggest differences in genotype at ga1 played a major role in cross-fertilization of populations differing in their genotype at the ga1 locus. Incompatibility may be selected effectively over different environments and the Ga1-s system may be of value to reduce cross-fertilization with GM corn pollen.