Effects of digestive status on the reptilian gut

Reptiles, including the Burmese python, Python molurus bivittatus, that feed at infrequent intervals show a prominent increase in gastrointestinal mass, metabolism and brush border transport rates after feeding. Current knowledge and theories around these phenomena, as well as studies on the innervation of the reptilian gut, are summarised in this review. Little is known about the putative changes in the nervous and humoral control systems of the gut, and it is not known whether feeding affects innervation and motility of the stomach and intestine. Using immunohistochemistry, we have investigated possible up/down regulation of several neurotransmitters in specimens that had been fasted for a minimum of 3 weeks and specimens that had ingested a large meal 2 days before the experiments were conducted. There were no major changes in the innervation by nerves containing calcitonin gene-related peptide (CGRP), galanin, nitric oxide synthase (NOS), pituitary adenylate cyclase-activating polypeptide (PACAP), somatostatin (SOM), substance P/neurokinin A (SP/NKA), or vasoactive intestinal polypeptide (VIP)-like immunoreactivity. Nor did we find any differences in the effect of substance P (stomach and intestine), galanin (intestine), or bradykinin (intestine) on motility in strip preparations from the gut wall. A significant increase in dry weight of the intestine was obtained 48 h after feeding. We conclude that although there are considerable changes in gut thickness and absorptive properties after feeding, the smooth muscle and its control appear little affected.     

Leaf water relations characteristics of differently potassium fertilized and watered field grown barley plants

Seasonal leaf water relations characteristics were studied in fully irrigated spring barley (Hordeum distichum L. cv. Gunnar) fertilized at low (50 kg K ha⁻¹) or high (200 kg K ha⁻¹) levels of potassium applied as KCl. The investigation was undertaken from about 14 days before anthesis until the milk ripe stage in leaves of different position and age. Additionally, the effects of severe water stress on leaf water relations were studied in the middle of the grain filling period in spring barley (cv. Alis). The leaf water relations characteristics were determined by the pressure volume (PV) technique. Water relations of fully irrigated plants were compared in leaf No 7 with the water relations of slowly droughted plants (cv. Alis). Leaf osmotic potential at full turgor (ψ _π ¹⁰⁰ ) decreased 0.1 to 0.3 MPa in droughted leaves indicating a limited osmotic adjustment due to solute accumulation. The leaf osmotic potential at zero turgor (ψ _π ⁰ ) was about −2.2 MPa in fully irrigated plants and −2.6 MPa in droughted plants. The relative water content at zero turgor (R⁰) decreased 0.1 unit in severely droughted leaves. The ratio of turgid leaf weight to dry weight (TW/DW) tended to be increased by drought. The tissue modulus of elasticity (ε) decreased in droughted plants and together with osmotic adjustment mediated turgor maintenance during drought. A similar response to drought was found in low and high K plants except that the R⁰ and ε values tended to be higher in the high K plants. Conclusively, during drought limited osmotic adjustment and increase in elasticity of the leaf tissue mediated turgor maintenance. These effects were only slightly modified by high potassium application. The seasonal analysis in fully irrigated plants (cv. Gunnar) showed that within about 14 days from leaf emergence ψ _π ¹⁰⁰ decreased from about −0.9 to −1.6 MPa in leaf No 7 (counting the first leaf to emerge as number one) and from about −1.1 to −1.9 MPa in leaf No 8 (the flag leaf) due to solute accumulation. A similar decrease took place in ψ _π ⁰ except that the level of ψ _π ⁰ was displaced to a lower level of about 0.2 to 0.3 MPa. Both ψ _π ¹⁰⁰ and ψ _π ⁰ tended to be 0.05 to 0.10 MPa lower in high K than in low K plants. R⁰ was about 0.8 to 0.9 and was independent of leaf position and age, but tended to be highest in high K plants. The TW/DW ratio decreased from about 5.5 in leaf No 6 to 4.5 in leaf No 7 and 3.8 in leaf No 8. The TW/DW ratio was 4 to 10% higher in high K than in low K plants indicating larger leaf cell size in the former. The apoplastic water content (V_a) at full turgor constituted about 15% in leaf No 7. ε was maximum at full turgor and varied from about 11 to 34 MPa. ε tended to be higher in high K plants. Conclusively, in fully watered plants an ontogenetically determined accumulation of solutes (probably organic as discussed) occurred in the leaves independent of K application. The main effect of high K application on water relations was an increase in leaf water content and a slight decrease in leaf ψ_π. The effect of K status on growth and drought resistance is discussed.     

Transfected HEK293 Cells Expressing Functional Recombinant Intercellular Adhesion Molecule 1 (ICAM-1) – A Receptor Associated with Severe Plasmodium falciparum Malaria

Intercellular adhesion molecule 1 (ICAM-1) is a membrane-bound glycoprotein expressed on endothelial cells and cells of the immune system. Human ICAM-1 mediates adhesion and migration of leucocytes, and is implicated in inflammatory pathologies, autoimmune diseases and in many cancer processes. Additionally, ICAM-1 acts as receptor for pathogens like human rhinovirus and Plasmodium falciparum malaria parasites. A group of related P. falciparum erythrocyte membrane protein 1 (PfEMP1) domains, the DBLβ, mediates ICAM-1 binding of P. falciparum-infected erythrocytes. This ICAM‑1-binding phenotype has been suggested to be involved in the development of cerebral malaria. However, more studies identifying cross-reactive antibody and ICAM-1-binding epitopes and the establishment of a clinical link between DBLβ expression and e.g. cerebral malaria are needed before the DBLβ domains can be put forward as vaccine candidates and go into clinical trials. Such studies require availability of functional recombinant ICAM-1 in large quantities. In this study, we compared recombinant ICAM-1 expressed in HEK293 and COS-7 cells with mouse myeloma NS0 ICAM-1 purchased from a commercial vendor in terms of protein purity, yield, fold, ability to bind DBLβ, and relative cost. We present a HEK293 cell-based, high-yield expression and purification scheme for producing inexpensive, functional ICAM‑1. ICAM-1 expressed in HEK293 is applicable to malaria research and can also be useful in other research fields.     

Promoting proteomics knowledge in Europe: report on the activities of the EuPA Education Committee 2006-2007

The early transition of knowledge from highly specialised and sophisticated proteomics research to a diverse community in need of know-how is a challenge that requires backing from advanced research centres and groups, and a coordinating body for the dissemination of this knowledge. The European Proteomics Association (EuPA) Education Committee signified this as a priority area when the EuPA was formed, and began its program to coordinate proteomics training and knowledge dissemination in 2006. This report serves as an update of our past activities and an announcement of upcoming events. Over the last year the EuPA Education Committee has coordinated or supported different educational activities including basic and advanced courses, a summer school, workshops and tutorials. A new programme of basic courses dubbed "Teaching the Teachers" has been initiated. These courses reach a larger, Europe wide, audience in a short timeframe, thus improving the opportunities for trainees of elementary proteomics techniques. Another important event has been the merger of the EuPA and HUPO (Human Proteome Organisation) Education Committees into a single one in order to combine ideas and ef for ts that will favour global education in proteomics.     

Denitrification, dissimilatory reduction of nitrate to ammonium, and nitrification in a bioturbated estuarine sediment as measured with N and microsensor techniques

Nitrogen and oxygen transformations were studied in a bioturbated (reworked by animals) estuarine sediment (Norsminde Fjord, Denmark) by using a combination of N isotope (NO(3)), specific inhibitor (C(2)H(2)), and microsensor (N(2)O and O(2)) techniques in a continuous-flow core system. The estuarine water was NO(3) rich (125 to 600 muM), and NO(3) was consistently taken up by the sediment on the four occasions studied. Total NO(3) uptake (3.6 to 34.0 mmol of N m day) corresponded closely to N(2) production (denitrification) during the experimental steady state, which indicated that dissimilatory, as well as assimilatory, NO(3) reduction to NH(4) was insignificant. When C(2)H(2) was applied in the flow system, denitrification measured as N(2)O production was often less (58 to 100%) than the NO(3) uptake because of incomplete inhibition of N(2)O reduction. The NO(3) formed by nitrification and not immediately denitrified but released to the overlying water, uncoupled nitrification, was calculated both from NO(3) dilution and from changes in NO(3) uptake before and after C(2)H(2) addition. These two approaches gave similar results, with rates ranging between 0 and 8.1 mmol of N m day on the four occasions. Attempts to measure total nitrification activity by the difference between NH(4) fluxes before and after C(2)H(2) addition failed because of non-steady-state NH(4) fluxes. The vertical distribution of denitrification and oxygen consumption was studied by use of N(2)O and O(2) microelectrodes. The N(2)O profiles measured during the experimental steady state were often irregularly shaped, and the buildup of N(2)O after C(2)H(2) was added was much too fast to be described by a simple diffusion model. Only bioturbation by a dense population of infauna could explain these observations. This was corroborated by the relationship between diffusive and total fluxes, which showed that only 19 to 36 and 29 to 62% of the total O(2) uptake and denitrification, respectively, were due to diffusion-reaction processes at the regular sediment surface, excluding animal burrows.     

The patchwork nature of rolling-circle plasmids: comparison of six plasmids from two distinct Bacillus thuringiensis serotypes

Bacillus thuringiensis, the entomopathogenic bacteria from the Bacillus cereus group, harbors numerous extrachromosomal molecules whose sizes vary from 2 to more than 200kb. Apart from the genes coding for the biopesticide delta-endotoxins located on large plasmids, little information has been obtained on these plasmids and their contribution to the biology of their host. In this paper, we embarked on a detailed comparison of six small rolling-circle replicating (RCR) plasmids originating from two major B. thuringiensis strains. The complete nucleotide sequences of plasmid pGI1, pGI2, pGI3, pTX14-1, pTX14-2, and pTX14-3 have been obtained and compared. Replication functions, comprising, for each plasmid, the gene encoding the Rep-protein, double-strand origin of replication (dso), single-strand origin of replication (sso), have been identified and analyzed. Two new families, or homology groups, of RCR plasmids originated from the studies of these plasmids (Group VI based on pGI3 and Group VII based on pTX14-3). On five of the six plasmids, loci involved in conjugative mobilization (Mob-genes and origin of transfer (oriT)) were identified. Plasmids pTX14-1, pTX14-2, and pTX14-3 each harbor an ORF encoding a polypeptide containing a central domain with repetitive elements similar to eukaryotic collagen (Gly-X-Y triplets). These genes were termed bcol for Bacillus-collagen-like genes.     

Variations in abundance of 2 repetitive sequences in Leymus and Psathyrostachys species.

The Ns genome of the genus Psathyrostachys is a component of the polyploid genome in the genus Leymus. Using fluorescence in situ hybridization (FISH), the occurrence and abundance of 2 tandem repetitive sequences from Leymus racemosus (Lam.) Tzvelev, pLrTaiI-1 (TaiI family) and pLrPstI-1 (1 class of 350-bp family), were assayed in 4 species of the genera Psathyrostachys and Leymus. The pLrPstI-1 sequence was absent in all 4 Psathyrostachys species. While P. fragilis and P. huashanica did not have the pLrTaiI-1 sequence, 15 accessions of P. juncea and 2 accessions of P. lanuginosa had pLrTaiI-1 sites ranging in number from 7 to 16 and from 2 to 21, respectively. The numbers of pLrTaiI-1 and pLrPstI-1 sites were 1-24 and 0-30, respectively, in L. ramosus; 2-31 and 5-36 in L. racemosus; 0-4 and 0 in L. mollis; 2-9 and 24-27 in L. secalinus. The FISH assay on pLrTaiI-1 was successfully converted to a sequence-tagged-site polymerase chain reaction (STS-PCR) test using a primer pair designed from the sequence of this repetitive DNA. Seventy-three accessions representing 27 Leymus species were assayed for the abundance of pLrTaiI-1 by STS-PCR. With a few exceptions of uniformity in some accessions, nearly all Leymus species observed were heterogeneous for the abundance of pLrTaiI-1 sequence and no Leymus species was totally devoid of this repetitive sequence. These findings may have significance for the understanding of phylogeny, nature of polyploidy, adaptive ranges, and breeding potential of Leymus species.