Production of concentrated freeze-dried cultures of Bifidobacterium longumin k-carrageenan-locust bean gum gel

Bifidobacterium longum was immobilized in k-carrageenan/locust bean gum gel beads, and cultured in a medium containing Lactobacillus MRS broth and whey-permeate. The same beads were incubated for 5 successive batch fermentations and freeze-dried following mixing with a protective solution. Viable population in the beads increased from 8 3 10 7 to 4.7 3 10 10 cfu/g after three batch fermentations, but no further increase in viable cell population could be achieved in the last two fermentations. The freeze-dried culture contained 3 3 10 10 cfu/g with a survival rate of approximately 10%. Survival to freeze-drying of immobilized cells was as good as that of classical free-cell cultures. Stability of freeze-dried cultures during storage at minus 17, 4 and 20°C was not influenced by immobilization.     

Hypothesis: Iron chelation plays a vital role in neutrophilic inflammation

Neutrophil influx into tissues occurs in many diverse diseases and can be associated with both beneficial and injurious effects. We hypothesize that the stimulus for certain neutrophilic inflammatory responses can be reduced to a series of competing reactions for iron, with either a labile or reactive coordination site available, between host chelators and chelators not indigenous to that specific living system. The iron focuses the transport of host phagocytic cells through a metal catalyzed generation of oxidant sensitive mediators including cytokines and eicosanoids. Many of these products are chemotactic for neutrophils. We also postulate that the iron increases the activity of the phagocyte associated NADPH oxidoreductase in the neutrophil. The function of this enzyme is likely to be the generation of superoxide in the hostÕs attempt to chemically reduce and dislodge the iron from its chelate complex. After the reoxidation of Fe in an aerobic environment, Fe will be coordinated by host lactoferrin released by the neutrophil. When complexed by this glycoprotein, the metal does not readily undergo oxidation/reduction and is safely transported to the macrophages of the reticuloendothelial system where it is stored in ferritin. Finally, we propose that the neutrophil will attempt to destroy the chelator not indigenous to the host by releasing granular contents other than lactoferrin. Inability to eliminate the chelator allows this sequence to repeat itself, which can lead to tissue injury. Such persistence of a metal chelate in the host may be associated with biomineralization, fibrosis, and cancer.     

Maintenance of transposable element copy number in natural populations of Drosophila melanogaster and D. simulans

To investigate the main forces controlling the containment of transposable elements (TE) in natural populations, we analyzed the copia, mdg1, and 412 elements in various populations of Drosophila melanogaster and D. simulans. A lower proportion of insertion sites on the X chromosome in comparison with the autosomes suggests that selection against the detrimental effects of TE insertions is the major force containing TE copies in populations of Drosophila. This selection effect hypothesis is strengthened by the absence of the negative correlation between recombination rate and TE copy number along the chromosomes, which was expected under the alternative ectopic exchange model (selection against the deleterious rearrangements promoted by recombination between TE insertions). A cline in 412 copy number in relation to latitude was observed among the natural populations of D. simulans, with very high numbers existing in some local populations (around 60 copies in a sample from Canberra, Australia). An apparent absence of selection effects in this Canberra sample and a value of transposition rate equal to 1–2 × 10^-3 whatever the population and its copy number agree with the idea of recent but temporarily drastic TE movements in local populations. The high values of transposition rate in D. simulans clearly disfavor the hypothesis that the low amount of transposable elements in this species could result from a low transposition rate. This revised version was published online in August 2006 with corrections to the Cover Date.     

Sensitivity to stress in the bivalve Macoma balthica from the most northern (Arctic) to the most southern (French) populations: low sensitivity in Arctic populations because of genetic adaptations?

The stress sensitivity, determined in copper exposureexperiments and in survival in air tests, and thegenetic structure, measured by means of isoenzymeelectrophoresis, were assessed in populations of theBaltic clam Macoma balthica (L.) from itssouthern to its northern distribution limit, in orderto test the hypotheses that near the distributionlimit the clams would be more stress sensitive andwould have a lower genetic variability. Thepopulations in west and north Europe show a stronggenetic resemblance. The populations in the sub-ArcticWhite Sea are genetically slightly different, and showa low stress sensitivity. The populations in theArctic Pechora Sea are genetically very distant fromthe other populations, and show the lowest stresssensitivity. Near the southern distribution limit, inagreement with the hypotheses, genetic variability islow and stress sensitivity high. On the other hand, incontrast to expectation, near the northerndistribution limit, in the populations of the PechoraSea, the genetic variability was higher, thus notreduced, and the stress sensitivity was low comparedto all other populations. Yet, it remains a questionif such is due to gradual physiologicalacclimatization (and ongoing differential selection)or to genetic adaptation.     

The Human Obesity Gene Map: The 1996 Update

An update of the human obesity gene map up to October 1996 is presented. Evidence from Mendelian disorders exhibiting obesity as a clinical feature, single-gene mutation rodent models, quantitative trait loci uncovered in crossbreeding experiments with mouse, rat, and pig models, association and case-control studies with candidate genes, and linkage studies with genes and other markers is reviewed. All chromosomal locations of the animal loci are converted into human genome locations based on syntenic relationships between the genomes. A complete listing of all these loci reveals that only 4 of the 24 human chromosomes are not yet represented, i.e., 9, 18, 21, and Y. Several chromosome arms are characterized by the presence of several putative loci. The following arms include at least three such loci: 1p, 1q, 3p, 4q, 6p, 7q, 8p, 8q, 11p, 11q, 15q, 20q, and Xq. Studies with negative association and linkage results are also reviewed.     

Cold-sensitive conditional mutations in Era, an essential Escherichia coli GTPase, isolated by localized random polymerase chain reaction mutagenesis

Abstract Conditional cold-sensitive mutations in Era, an essential Escherichia coli GTPase, were isolated. Localized random polymerase chain reaction (PCR) mutagenesis employing Taq and T7 DNA polymerases under error prone amplification conditions was exploited to generate mutations in the era gene. A plasmid exchange technique was used to identify conditional cold-sensitive mutations in Era that give rise to defective cell growth below 30 °C. Three recessive missense mutations in Era, N26S, A156D, and E200K, were isolated. All three mutations are located at residues conserved in Era homologues from Streptococcus mutans and Coxiella burnetii .     

Calmodulin during development and metamorphosis in urodelan amphibians

Calmodulin isolated and purified to homogeneity from young larvae is very similar to that obtained from adult Pleurodeles waltlii and these proteins are almost identical to previously described vertebrate calmodulins. During P. waltlii development, an increase in total individual calmodulin content is observed after the heart beating stage. In dorsal axial muscle, calmodulin level which is very high at the beginning of larval life (premetamorphosis) decreases strikingly in the first part of prometamorphosis. Such an evolution is observed in Ambystoma mexicanum too. Then, a significant increase occurs during metamorphosis. In contrast, calmodulin level in P. waltlii cardiac ventricular muscle increases continuously from hatching to the end of metamorphic climax. Thyroxine treatment which promotes precocious metamorphosis in P. waltlii and experimental metamorphosis in neotenic A. mexicanum, induces a rapid and significant increase in muscle calmodulin concentration.     

An immunological approach to quantitate RNA polymerases in plant cell extracts

A polyclonal antiserum was raised against highly purified RNA polymerase II from soybean embryos. Pure RNA polymerase II was fractionated on sodium dodecyl sulfate-polyacrylamide gels and transferred onto nitrocellulose sheets, incubated with the immune antiserum and then with iodinated protein A. Autoradiograms showed that the immune antiserum recognized all subunits of RNA polymerase II. Subunits 42, 27 and 16 kdalton were particularly reactive. Application of this transfer technique to protein extracts from soybean embryos or from cultured soybean cells allowed the identification of subunits of RNA polymerase II in the extracts. Analysis of the staining of the bands on the autoradiograms for increasing amounts of pure RNA polymerase II demonstrated that the transfer was quantitative, so that standard curves could be drawn to estimate the unknown amounts of enzyme in the extracts.Abbreviations DEAE diethylaminoethyl - SDS sodium dodecyl sulfate     

Chitin synthetase ofNeurospora crassa: Inhibition by nikkomycin,polyoxin B,and UDP

The inhibitory effects of nikkomycin, polyoxin B, and UDP were tested on particulate chitin synthetase activity (UDP-2-acetamido-2-deoxy-D-glucose: chitin-4-B-acetamidodeoxy-D-glucosyltransferase, E.C.2.4.1.16) fromNeurospora crassa. Two approaches were used: (a) inhibitors were tested for their individual effects on chitin synthetase activity; (b) paired combinations of inhibitors were examined to establish whether the compounds affected inhibition by binding at the same enzyme site. The first method showed that the three compounds are linear competitive inhibitors, i.e. each competes directly with the substrate for binding. K_{i app} values were: UDP, 0.8 mM; polyoxin B, 32 M; and nikkomycin, 2 M. The second method showed that the inhibitors compete with each other for binding; thus they bind at the same site. The pyrimidine nucleoside moiety of these inhibitors is an essential component for effective inhibition, but the potency of inhibition is critically dependent on the conformation of the side group attached to carbon 5 of the ribose sugar.