Use of a Granulocyte Immunofluorescence Assay Designed for Humans for Detection of Antineutrophil Cytoplasmic Antibodies in Dogs with Chronic Enteropathies

Perinuclear antineutrophil cytoplasmic antibodies (pANCA) previously have been shown to be serum markers in dogs with chronic enteropathies, with dogs that have food‐responsive disease (FRD) having higher frequencies of seropositivity than dogs with steroid‐responsive disease (SRD). The indirect immunofluorescence (IIF) assay used in previous publications is time‐consuming to perform, with low interobserver agreement. Forty‐four dogs with FRD, 20 dogs with SRD, 20 control dogs, and 38 soft‐coated wheaten terrier (SCWT) or SCWT‐cross dogs

Single copy shRNA configuration for ubiquitous gene knockdown in mice

RNA interference through the expression of small hairpin RNA (shRNA) molecules has become a very promising tool in reverse mouse genetics as it may allow inexpensive and rapid gene function analysis in vivo. However, the prerequisites for ubiquitous and reproducible shRNA expression are not well defined. Here we show that a single copy shRNA-transgene can mediate body-wide gene silencing in mice when inserted in a defined locus of the genome. The most commonly used promoters for shRNA expression, H1 and U6, showed a comparably broad activity in this configuration. Taken together, the results define a novel approach for efficient interference with expression of defined genes in vivo. Moreover, we provide a rapid strategy for the production of gene knockdown mice combining recombinase mediated cassette exchange and tetraploid blastocyst complementation approaches

Human amniotic fluid glycoproteins expressing sialyl Lewis carbohydrate antigens stimulate progesterone production in human trophoblasts in vitro

Background: Progesterone is thought to mediate immune modulator effects by regulating uterine responsiveness. The aim of the study was to clarify the effect of transferrin and glycodelin A (former name PP14) as sialyl Lewis X-expressing glycoproteins on the release of progesterone by trophoblast cells in vitro. Methods: Cytotrophoblast cells were prepared from human term placentas by standard dispersion of villous tissue followed by a Percoll gradient centrifugation step. Trophoblasts were incubated with varying concentrations (50-300 mug/ml) of human amniotic fluid- and serum-transferrin as well as with glycodelin A. Culture supernatants were assayed for progesterone, human chorionic gonadotropin (hCG) and cortisol by enzyme immunometric methods. Results: The release of progesterone is increased in amniotic fluid transferrin- and glycodelin A-treated trophoblast cell cultures compared to untreated trophoblast cells. There is no relation between transferrin and the hCG or cortisol production of trophoblast cells. Conclusion: The results suggest that sialyl Lewis carbohydrate antigen-expressing amniotic fluid glycoproteins modulate the endocrine function of trophoblasts in culture by upregulating progesterone production. Copyright (C) 2004 S. Karger AG, Basel

Development and bioorthogonal activation of palladium-labile prodrugs of gemcitabine

Bioorthogonal chemistry has become one of the main driving forces in current chemical biology, inspiring the search for novel biocompatible chemospecific reactions for the past decade. Alongside the well-established labeling strategies that originated the bioorthogonal paradigm, we have recently proposed the use of heterogeneous palladium chemistry and bioorthogonal Pd⁰-labile prodrugs to develop spatially targeted therapies. Herein, we report the generation of biologically inert precursors of cytotoxic gemcitabine by introducing Pd⁰-cleavable groups in positions that are mechanistically relevant for gemcitabine’s pharmacological activity. Cell viability studies in pancreatic cancer cells showed that carbamate functionalization of the 4-amino group of gemcitabine significantly reduced (>23-fold) the prodrugs’ cytotoxicity. The <i>N</i>-propargyloxycarbonyl (<i>N</i>-Poc) promoiety displayed the highest sensitivity to heterogeneous palladium catalysis under biocompatible conditions, with a reaction half-life of less than 6 h. Zebrafish studies with allyl, propargyl, and benzyl carbamate-protected rhodamines confirmed <i>N</i>-Poc as the most suitable masking group for implementing <i>in vivo</i> bioorthogonal organometallic chemistry

Dry and wet deposition of nutrients from the tropical Atlantic atmosphere: Links to primary productivity and nitrogen fixation

Atmospheric deposition fluxes of soluble nutrients (N, P, Si, Fe, Co, Zn) to the tropical North Atlantic were determined during cruise M55 of the German SOLAS programme. Nutrient fluxes were highest in the east of the section along 10°N, owing to the proximity of source regions in West Africa and Europe, and lowest in the west, for both dry and wet deposition modes. In common with other recent studies, atmospheric P and Si inputs during M55 were strongly depleted relative to the stoichiometry of phytoplankton Fe, N, P and Si requirements. Atmospheric N inputs were equivalent to 0.1–4.7% of observed primary productivity during the cruise. Atmospheric nutrient supply was also compared to observed nitrogen fixation rates during M55. While atmospheric Fe supply may have been sufficient to support N fixation (depending on the relationship between our simple Fe leaching experiment and aerosol Fe dissolution in seawater), atmospheric P supply was well below the required rate. The stable nitrogen isotope composition of nitrate–N in aerosol and rain was also determined. Results of a simple model indicate that atmospheric deposition and nitrogen fixation introduce similar amounts of isotopically light nitrogen into surface waters of the study region. This implies that nitrogen isotope-based methods would overestimate nitrogen fixation here by a factor of 2, if atmospheric inputs were not taken into account

Republican Journal: Vol. 57, No. 50 - December 10,1855

https://digitalmaine.com/rj_1885/1050/thumbnail.jp

Synthesis, Characterization, and Bioactivity of the Photoisomerizable Tubulin Polymerization Inhibitor azo-Combretastatin A4

Combretastatin A4 is a stilbenoid tubulin binding mitotic inhibitor whose conformation greatly influences its potency, making it an excellent candidate for adaptation as a photoactivatable tool. Herein we report a novel synthesis, the facile isomerization with commercial grade equipment, and biological activity of azo-combretastatin A4 <i>in vitro</i> and in human cancer cells. Photoisomerized azo-combretestatin A4 is at least 200-fold more potent in cellular culture, making it a promising phototherapeutic and biomedical research tool