Fate and behaviour of a seed-applied Pseudomonas brassicacearum strain in a winter wheat field trial,as determined by analysis with SCAR markers

The fate and behaviour of the seed-applied biocontrol strain Pseudomonas brassicacearum MA250 in a field trial with winter wheat was determined using sequence-characterised amplified region (SCAR) markers. Samples of belowground plant parts from healthy and withered (due to snow mould infection) seedlings were collected approximately one and seven months after sowing, which was performed in early autumn. DNA was extracted from roots and remaining parts of seeds with adhering soil, and the abundance of the strain was determined in quantitative real-time PCR (qPCR) assays. The results show that the introduced strain persisted over the whole trial-period of seven months. On termination of the trial (after seven months) the belowground plant parts of each plant housed 10⁶–10⁷ cells, substantially less than the original approximately 10⁹ cells inoculated onto the seed. In healthy seedlings, there was a shift in cell numbers from seeds to roots between the samplings, suggesting colonisation of the roots during this time. The results show that with sufficient attention given to analytical control measures and the possibility of resident background populations, SCAR markers in combination with qPCR provide valuable information regarding the fate and behaviour of biocontrol micro-organisms under field conditions.     

Cyclopenta[g]quinazoline-based antifolates: the effect of the chirality at the 6-position on the inhibition of thymidylate synthase (TS).

Cyclopenta[g]quinazoline-based inhibitors of thymidylate synthase (TS) possess a chiral centre at the 6-position of the molecule. The effect of this chirality on the inhibition of TS was investigated by synthesising compounds 6S-1a-c, 6R-1a-c. It was shown, in particular with the diastereoisomers 6S-1c, 6R-1c, that the inhibitory activity against TS is mainly due to the 6S diastereoisomer rather than the 6R diastereoisomer, which is virtually inactive.     

The influence of rinsing routines on fluoride retention after toothbrushing

Objective: An important determinant of the anticaries effect of brushing with a fluoride dentifrice is the retention of fluoride in the oral fluids. This study measured the influence of various post‐brushing rinsing routines on fluoride retention. Setting: The Faculty of Odontology, Göteborg University, Sweden. Subjects: Twenty‐one healthy dental students, aged 23±2 yr. Desing: A randomised, cross‐over protocol, comparing various post‐brushing rinsing procedures, with the following variables: volume of rinsing water (5 mL vs. 10×3mL) and duration of rinsing (10 vs. 60 s). Samples of whole saliva, interdental fluid and interdental plaque were analysed for fluoride content. Results: The duration of rinsing did not influence fluoride retention in whole saliva, interdental fluid or interdental plaque. Rinsing with a higher volume of water substantially decreased fluoride retention in all samples. Conclusion: Brushing with a fluoride dentifrice should be followed by rinsing procedures which enhance the retention of fluoride. Rinsing with large volumes of water decreases fluoride retention in the oral fluids, and should be discouraged.     

The in vivo toxicity of carbon tetrachloride and carrageenan on heart microsomes: analysis by Fourier transform infrared spectroscopy

We investigated the sensitivity of rat heart microsomes to free radical attack using Fourier transform infrared (FT-IR) spectroscopy. This physico-chemical method seemed a valuable technique: quite sensitive to changes in the vibrational spectra. The spectral variations observed between normal and treated rats were in great part due to reactive oxygen species that led to changes in protein conformation involving beta-sheets, aggregation of proteins, and modification of protein synthesis. Carrageenan-induced inflammation slightly enhanced the total lipid content; rearrangement of acyl chains and accumulation of cholesterol esters and phospholipids also occurred in the treated rats. Carbon tetrachloride induced a decrease in both lipid and protein contents. The level of glucidic substrates was diminished with carbon tetrachloride and enhanced with carrageenan; these changes were due to metabolic interactions between cell components and drugs. FT-IR spectroscopy provided an accurate means to monitor, in rat heart, the in vivo effects of inflammatory and peroxidative damages, to discriminate and classify the affected cells, and to correlate the findings with known physiological and biochemical data in close relationship with metabolic disruptions induced by the two xenobiotics.     

Fourier transform IR spectroscopic appraisal of radiation damage in Micrococcus luteus

Fourier transform IR spectroscopy (FTIR) is used to analyze cells of Micrococcus luteus, the type species of the highly heterogeneous genus Micrococcus that belongs to the Micrococcaceae family. The cells of M. luteus, which is a Gram-positive and yellow-pigmented bacterium, are submitted to increasing doses of gamma radiation. Irradiation leads to the generation of reactive oxygen species that induce biochemical changes as shown in spectral profiles. Beyond a dose of 0.70 kGy, significant differences between samples are observed, particularly in the 1485-900 cm(-1) region, which contains information about membrane lipids, cell wall polysaccharides, and nucleic acids. After a dose of 16.50 kGy, M. luteus is reincubated for times ranging from 1 to 24 h. Postirradiation reincubated bacteria are found far from the control and irradiated cells (mainly in the 985-900 cm(-1) range), suggesting that a biomolecular rearrangement occurs as soon as reincubation begins in the growth medium. Thus, FTIR spectroscopy appears to be a very useful technique for the rapid visualization of the alterations induced by both the radiation and mutagenic response during reincubation. The use of mathematical methods gives good insight into the biomolecular compounds involved in these two mechanisms. In view of these preliminary results, we hypothesize that it can be successfully applied to any type of tissue and that it may be a future interesting tool for evaluating the effects of radiation in humans.     

Assignment of the gene locus for severe congenital neutropenia to chromosome 1q22 in the original Kostmann family from Northern Sweden

Autosomal recessive severe congenital neutropenia (SCN) or Kostmann syndrome is characterised by reduced neutrophil counts and subsequent recurrent bacterial infections. The disease was originally described in a large consanguineous pedigree from Northern Sweden. A genome-wide autozygosity scan was initiated on samples from four individuals in the original pedigree using high density single nucleotide polymorphism (SNP) genotyping arrays in order to map the disease locus. Thirty candidate regions were identified and the ascertainment of samples from two additional patients confirmed a single haplotype with significant association to the disorder (p<0.01) on chromosome 1q22. One affected individual from the original Kostmann pedigree was confirmed as a phenocopy. The minimal haplotype shared by affected individuals spans a candidate region of 1.2 Mb, containing several potential candidate genes.     

Endosomal SNARE proteins regulate CFTR activity and trafficking in epithelial cells

The Cystic Fibrosis Transmembrane conductance Regulator (CFTR) protein is a chloride channel localized at the apical plasma membrane of epithelial cells. We previously described that syntaxin 8, an endosomal SNARE (Soluble N-ethylmaleimide-sensitive factor Attachment protein REceptor) protein, interacts with CFTR and regulates its trafficking to the plasma membrane and hence its channel activity. Syntaxin 8 belongs to the endosomal SNARE complex which also contains syntaxin 7, vti1b and VAMP8. Here, we report that these four endosomal SNARE proteins physically and functionally interact with CFTR. In LLC-PK1 cells transfected with CFTR and in Caco-2 cells endogenously expressing CFTR, we demonstrated that endosomal SNARE protein overexpression inhibits CFTR activity but not swelling- or calcium-activated iodide efflux, indicating a specific effect upon CFTR activity. Moreover, co-immunoprecipitation experiments in LLC-PK1-CFTR cells showed that CFTR and SNARE proteins belong to a same complex and pull-down assays showed that VAMP8 and vti1b preferentially interact with CFTR N-terminus tail. By cell surface biotinylation and immunofluorescence experiments, we evidenced that endosomal SNARE overexpression disturbs CFTR apical targeting. Finally, we found a colocalization of CFTR and endosomal SNARE proteins in Rab11-positive recycling endosomes, suggesting a new role for endosomal SNARE proteins in CFTR trafficking in epithelial cells.