Depletion of preexisting B-cell lymphoma 2-expressing senescent cells before vaccination impacts antigen-specific antitumor immune responses in old mice

The age-related decline in immunity reduces the effectiveness of vaccines in older adults. Immunosenescence is associated with chronic, low-grade inflammation, and the accumulation of senescent cells. The latter express Bcl-2 family members (providing resistance to cell death) and exhibit a pro-inflammatory, senescence-associated secretory phenotype (SASP). Preexisting senescent cells cause many aging-related disorders and therapeutic means of eliminating these cells have recently gained attention. The potential consequences of senescent cell removal on vaccine efficacy in older individuals are still ignored. We used the Bcl-2 family inhibitor ABT-263 to investigate the effects of pre-vaccination senolysis on immune responses in old mice. Two different ovalbumin (OVA)-containing vaccines (containing a saponin-based or a CpG oligodeoxynucleotide adjuvant) were tested. ABT-263 depleted senescent cells (apoptosis) and ablated the basal and lipopolysaccharide-induced production of SASP-related factors in old mice. Depletion of senescent cells prior to vaccination (prime/boost) had little effect on OVA-specific antibody and T-cell responses (slightly reduced and augmented, respectively). We then used a preclinical melanoma model to test the antitumor potential of senolysis before vaccination (prime with the vaccine and OVA boost by tumor cells). Surprisingly, ABT-263 treatment abrogated the vaccine's ability to protect against B16 melanoma growth in old animals, an effect associated with reduced antigen-specific T-cell responses. Some, but not all, of the effects were age-specific, which suggests that preexisting senescent cells were partly involved. Hence, depletion of senescent cells modifies immune responses to vaccines in some settings and caution should be taken when incorporating senolytics into vaccine-based cancer therapies.     

Conservation of Dynamics Associated with Biological Function in an Enzyme Superfamily

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Three new naraoiid species from the Burgess Shale,with a morphometric and phylogenetic reinvestigation of Naraoiidae

Naraoiids are non‐biomineralized euarthropods characterized by the complete fusion of post‐cephalic tergo‐pleurae into a single shield, as well as an extensively ramified digestive tract. Ranging from the early Cambrian to the late Silurian (Pridoli), these arthropods of simple appearance have traditionally been associated with the early diversification of trilobites and their close relatives, but the interrelationships and affinities of naraoiids within Artiopoda remain poorly characterized. Three new species from the Burgess Shale (middle Cambrian, Stage 5) of British Columbia, Canada, are described here: Misszhouia canadensis sp. nov., from Marble Canyon (Kootenay National Park), the first species belonging to the genus Misszhouia outside of China; Naraoia magna sp. nov., from Marble Canyon and also from the Raymond Quarry (Yoho National Park), the largest species of Naraoia described thus far, reaching up to 9 cm in length; and Naraoia arcana sp. nov., from two sublocalities on Mount Stephen (Yoho National Park), defined by its unusual combination of spines. This new material shows that gut morphology is no longer a reliable character to distinguish Misszhouia from Naraoia. We demonstrate that Naraoia and Misszhouia can instead be discriminated morphometrically, based on simple metrics of the dorsal exoskeleton. Our quantitative results also help with inter‐specific discrimination and illustrate possible cases of sexual dimorphism. Phylogenetically, the inclusion of morphometric data adds resolution to our cladogram, although parsimony and likelihood treatments provide somewhat different evolutionary scenarios. In all cases, liwiines are nested within Naraoiidae, resolved as the most derived clade of trilobitomorph arthropods.     

Multiscale optical flow computation from the monogenic signal

We have developed an algorithm for the estimation of cardiac motion from medical images. The algorithm exploits monogenic signal theory, recently introduced as an N-dimensional generalization of the analytic signal. The displacement is computed locally by assuming the conservation of the monogenic phase over time. A local affine displacement model replaces the standard translation model to account for more complex motions as contraction/expansion and shear. A coarse-to-fine B-spline scheme allows a robust and effective computation of the models parameters and a pyramidal refinement scheme helps handle large motions. Robustness against noise is increased by replacing the standard pointwise computation of the monogenic orientation with a more robust least-squares orientation estimate. This paper reviews the results obtained on simulated cardiac images from different modalities, namely 2D and 3D cardiac ultrasound and tagged magnetic resonance. We also show how the proposed algorithm represents a valuable alternative to state-of-the-art algorithms in the respective fields.     

Genetic Study of Suppressor-Sensitive Mutants of the Bacillus subtilis Bacteriophage φ29

With bacteriophage phi29 of Bacillus subtilis 133, suppressor-sensitive (sus) hydroxylamine mutants have been isolated. Intracistronic and intercistronic quantitative complementation placed the mutants in 13 cistrons, and three-factor crosses have been used to assign an unambiguous order for 10 cistrons. Recombination frequencies have been presented for several regions of the genome to facilitate comparison of the sus system with the previously published temperature-sensitive mapping systems.     

Purification of an N-acetyl-d-glucosamine specific lectin (P. B. A.) from epidermal cell membranes of Pieris brassicae L

We report the isolation and the purification of an N-acetyl-d-glucosamine specific lectin capable of agglutinating either fixed trypsinized rabbit erythrocytes or chitin particles. An agglutinin assay based on the affinity of this lectin for the chitin was devised with fluorescent particles of scorpion cuticle to measure lectin activity during purification steps.Lectin was isolated from epidermal cell membranes; its molecular weight was determined by gel filtration and polyacrylamide electrophoresis in sodium dodecyl sulfate. Mr was estimated to be 43,000. Lectin could be constituted by two subunits, Mr of which was estimated to be 23,000. The specificity of this lectin against N-acetyl-d-glucosamine and its oligomers suggests a possible role in the dynamics of these saccharides during the cuticle cycle.     

PCR-mediated screening and labeling of DNA from clones

A simplified and economical protocol for DNA library screening and nonradioactive labeling is described. Bacterial clones are lysed in 1% of Triton X-100 and subjected to polymerase chain reaction in the presence of digoxigenin-11-dUTP to screen and simultaneously to label the DNA inserts. Bacteriallysates are stable in storage at −20°C and can be used repeatedly for PCR-mediated labeling. In this protocol, very low concentrations of dNTP, digoxigenin-dUTP, and primers are used in combination with a reduced reaction volume. This will considerably reduce the expense of screening and labeling bacterial clones and facilitate the exchange of DNA probes among laboratories.     

Effect of transformation ofAzotobacter vinelandii with the low copy number plasmid pRK290

We previously observed that whenAzotobacter vinelandii was transformed by different broad-host-range plasmids, normal cellular functions such as growth and siderophore production are impaired. In the present work, whenA. vinelandii was transformed with the low copy number plasmid pRK290, the extent of this metabolic impairment was lessened, as evidenced by increased siderophore production and moderate levels of growth on medium that lacks added iron. It is concluded that the severity of the plasmid-induced metabolic load reflects the relative level of expression of plasmid-encoded proteins.     

Effective resolution of racemic pirlindole at the preparative scale

Pirlindole, a racemic antidepressant drug, was recently resolved using the derivatization method coupled with preparative HPLC. In order to improve this technique, the use of amino acid derivatives as chiral derivatizing agents (CDA) was investigated. Among different residues, the (L)-phenylalanine methyl ester was found to be very effective to separate pirlindole enantiomers using a medium pressure liquid chromatographic (MPLC) method. This procedure is better adapted to preparative application than HPLC. Thus, several grams of the pirlindole antipodes were isolated and characterized. These two enantiomers permitted the study of the stereochemical influence at the pharmacological level. Chirality 11:261–266, 1999. © 1999 Wiley-Liss, Inc.