Impaired mucus-bicarbonate barrier in Helicobacter pylori-infected mice

To resist the harsh intrinsic milieu, several lines of defense exist in the stomach. The aim of this study was to investigate the effect of the gastric pathogen Helicobacter pylori on these mechanisms in vivo. We used FVB/N mice expressing human alpha-1,3/4-fucosyl transferase (producing Lewis b epitopes) and inoculated with H. pylori 1. Mice were anesthetized with isoflurane or Hypnorm-midazolam, the stomach was exteriorized, and the surface of the corpus mucosa was exposed. Mucus thickness was measured with micropipettes, juxtamucosal pH (pH(jm)) was measured with pH-sensitive microelectrodes, blood flow was measured with laser-Doppler flowmetry, and mRNA levels of the bicarbonate transporter SLC26A9 were quantified with real-time PCR. The increase in mucosal blood flow seen in response to luminal acid (pH 1.5) in control animals (140 +/- 9% of control) was abolished in infected mice. The firmly adherent mucus layer was significantly thinner in infected mice (31 +/- 2 microm) than in control mice (46 +/- 5 microm), and no mucus accumulation occurred in infected mice. pH(jm) decreased significantly more on exposure to luminal acid in infected mice (luminal pH 1.5, pH(jm) 2.4 +/- 0.7) than in control mice (pH(jm) 6.4 +/- 0.5). Despite reduced pH(jm), SLC26A9 mRNA expression was significantly, by increased 1.9-fold, in infected mice. The reduction in pH(jm) by infection with H. pylori might be due to a reduced firmly adherent mucus layer, increased mucus permeability to H(+), and/or inhibition of bicarbonate transport. The upregulation of SLC26A9 in H. pylori-infected epithelium might be a result of continuous inhibition of the transporter, e.g., by ammonium, a H. pylori product, which has been previously shown to inhibit SLC26A9.     

ONTOGENY AND DIFFERENTIATION OF SCLEREIDS IN RAUWOLFIA

An interesting anatomic feature of Rauwolfia is the occurrence of a remarkable type of sclereid in the stem and root. The initials of the sclereids in the stem arise in the ground tissue element of the pith in a region between 50 and 70μ below the surface of the shoot apex. This region of the shoot remains surrounded by a whorl of either 3 or 4 leaves. Sclereids initiate in succession in association with each whorl of leaves. Thus, the sclereids are restricted to the nodes. The sclereids in the stem arise as a primary element of the shoot from the ground tissue of the pith. In the root, they differentiate from the cells of the phelloderm and are secondary in origin. Morphologically, the sclereids in these 2 organs are basically the same, except that the sclereids in the stem are larger in size than those in the root. A solitary cell, or 2 to several cells in a longitudinal cell file (originated from a single mother cell), may differentiate into sclereid initials. The growth of the sclereids through relatively compact ground tissue of the pith is possibly accomplished by a combination of gliding growth and apical intrusive process. The sclereid initials grow rapidly and force their way between the parenchymatous cells. As a result, the neighboring cells lose their original surface contacts. Sclereids increase in size rapidly, and, therefore, very enlarged sclereids with thin primary walls may be observed in the second node. They mature progressively in basipetal direction in the subjacent nodes. In the fifth or sixth node, mature sclereids with massive secondary walls are most common. The secondary walls of sclereids contain much lignin as determined by the phloroglucinol-HCl test. The walls of sclereids at maturity show a variable number of lamellae ranging from 10 to 15 in the lateral walls. A remarkable feature of the sclereids is their canal-like pits in the secondary walls. Two adjacent pits may coalesce uniquely to form a Y-like configuration directed centrifugally from the lumen of the sclereids. The sclereids are ventrically symmetrical, joined end-to-end by their transverse walls like 2 superimposed young fibers.     

Einiges über die kopulation bei aturus scaber und Midea orbiculata

Ohne Zusammenfassung     

Über den begattungsvorgang bei einigen arrhenurus-arten

Ohne Zusammenfassung     

POPDC1 scaffolds a complex of adenylyl cyclase 9 and the potassium channel TREK‐1 in heart

The establishment of macromolecular complexes by scaffolding proteins is key to the local production of cAMP by anchored adenylyl cyclase (AC) and the subsequent cAMP signaling necessary for cardiac functions. We identify a novel AC scaffold, the Popeye domain‐containing (POPDC) protein. The POPDC family of proteins is important for cardiac pacemaking and conduction, due in part to their cAMP‐dependent binding and regulation of TREK‐1 potassium channels. We show that TREK‐1 binds the AC9:POPDC1 complex and copurifies in a POPDC1‐dependent manner with AC9 activity in heart. Although the AC9:POPDC1 interaction is cAMP‐independent, TREK‐1 association with AC9 and POPDC1 is reduced upon stimulation of the β‐adrenergic receptor (βAR). AC9 activity is required for βAR reduction of TREK‐1 complex formation with AC9:POPDC1 and in reversing POPDC1 enhancement of TREK‐1 currents. Finally, deletion of the gene‐encoding AC9 (Adcy9) gives rise to bradycardia at rest and stress‐induced heart rate variability, a milder phenotype than the loss of Popdc1 but similar to the loss of Kcnk2 (TREK‐1). Thus, POPDC1 represents a novel adaptor for AC9 interactions with TREK‐1 to regulate heart rate control.     

Recombinant amyloid beta-peptide production by coexpression with an affibody ligand

Background

The isolation and production of human monoclonal antibodies is becoming an increasingly important pursuit as biopharmaceutical companies migrate their drug pipelines away from small organic molecules. As such, optimization of monoclonal antibody technologies is important, as this is becoming the new rate-limiting step for discovery and development of new pharmaceuticals. The major limitations of this system are the efficiency of isolating hybridoma clones, the process of stabilizing these clones and optimization of hybridoma cell secretion, especially for large-scale production. Many previous studies have demonstrated how perturbations in the aqueous environment can impact upon cell biology. In particular, radio frequency (RF) irradiation of solutions can have dramatic effects on behavior of solutions, cells and in particular membrane proteins, although this effect decays following removal of the RF. Recently, it was shown that nanoparticle doping of RF irradiated water (NPD water) produced a stabilized aqueous medium that maintained the characteristic properties of RF irradiated water for extended periods of time. Therefore, the ordering effect in water of the RF irradiation can now be studied in systems that required prolonged periods for analysis, such as eukaryotic cell culture. Since the formation of hybridoma cells involves the formation of a new membrane, a process that is affected by the surrounding aqueous environment, we tested these nanoparticle doped aqueous media formulations on hybridoma cell production.

Results

In this study, we tested the entire process of isolation and production of human monoclonal antibodies in NPD water as a means for further enhancing human monoclonal antibody isolation and production. Our results indicate an overall enhancement of hybridoma yield, viability, clonability and secretion. Furthermore, we have demonstrated that immortal cells proliferate faster whereas primary human fibroblasts proliferate slower in NPD water.

Conclusion

Overall, these studies indicate that NPD water can enhance cell proliferation, clonability and secretion. Furthermore, the results support the hypothesis that NPD water is effectively composed of stable microenvironments.     

Telomeres in the '80s: a few recollections

Elizabeth Blackburn, Carol Greider and Jack Szostak were recently recognized with the Lasker award for their work on telomerase and the role of this enzyme in cellular proliferation. Vicki Lundblad reflects on the excitement as these experiments were unfolding.     

Analysis of amino acids: neurochemical application

For high sensitivity analysis of neuroactive amino acids, liquid chromatography employing precolumn derivatisation with o-phthalaldehyde (OPA) is suitable for several reasons. The OPA reagent is non-fluorescent per se, the reaction occurs rapidly in alkaline aqueous solutions and forms highly fluorescent derivatives with primary amines.     

Isolation of the intermediate filament protein vimentin by chromatofocusing

A novel, simple and relatively rapid method is described for the isolation of the intermediate-sized filament protein vimentin from eye lens tissue. Chromatofocusing is applied as the sole purification step. The apparent isoelectric point of the protein in 6 M urea and at 22°C is 4.9. Electrophoretic mobility on one- and two-dimensional polyacrylamide gels, solubility in 6 M urea and amino acid composition were used for identification