162,088 research outputs found
Embryonic development and esterase activity of eggs of pieris brassicae in relation to tepp poisoning
No full textThe embryological development of eggs of Pieris brussicae was studied in relation to the occurrence of enzymes hydrolysing phenyl acetate and acetylcholine. Phenyl acetate is hydrolysed at a high rate at all stages of development of the embryo. Hydrolysis of acetylcholine only becomes appreciable in the later stages of development. The first significant level of hydrolysis of acetylcholine can be correlated with the development of a nervous system in the embryo to a stage where it may be functional. Aqueous solutions of TEPP were applied to eggs soon after they were laid. Low doses of TEPP allowed a high percentage of eggs to develop to the point of hatching before death occurred. Most fully developed embryos became active before they died. As the dose was increased less development took place and with very high doses little development occurred. The significance of these results is discussed. The available evidence does not indicate that the poison penetrates slowly nor that it is 'locked up' and later released. The explanation that seems to fit the evidence best is that the poison penetrates rapidly and reacts irreversibly with, probably phosphorylates, one or more components of the egg, the extent of subsequent development depending upon the proportion of a biochemical system or the number of systems inactivated. Whilst inhibition of cholinesterase may play a part in the poisoning process, at least under some conditions, the evidence indicates that the death of the embryo may result from some other cause
TepP/Ct875 is secreted at <i>Chlamydia</i> entry sites and is phosphorylated upon translocation.
No full text<p><b>A</b>) TepP is secreted from EBs early during infection. HeLa cells were infected with LGV-L2 at a MOI of 20 for 2 hr. Cells were fixed, permeabilized and immunostained with antibodies against <i>Chlamydia</i> LPS (red), Ct875/TepP (green) and Slc1 (green). Scale bar: 1 µm. <b>B</b>) Immunoblot analysis of total cell lysates of <i>Chlamydia</i> infected cells shows the appearance of at least three major tyrosine-phosphorylated proteins (>150 kDa, 100 kDa, and 65–70 kDa) within 1 hpi. <b>C</b>) TepP colocalizes with anti-phospho-Tyr signals at <i>Chlamydia</i> entry sites. HeLa cells were infected as in A) and immunostained with anti-TepP (green) and anti-p-Tyr (red) antibodies. Host and bacterial DNA were stained with DAPI (blue). Scale bar: 10 µm (upper panel); 2 µm (lower panel). <b>D</b>) TepP is phosphorylated at tyrosine residues upon EB association with host cells. Cells were infected as in B) with an MOI of 100 for the indicated times and TepP was immunoprecipitated from cell lysates and analyzed by immunoblotting with anti-TepP and anti p-Tyr antibodies. TepP was recognized by anti p-Tyr antibodies only upon association with host cells. <b>E–F</b>) TARP and TepP are tyrosine-phosphorylated at different rates during <i>Chlamydia</i> infection. HeLa cells were infected as in B) and total protein lysates were generated at 0, 5, 15, 30 min and 1 hpi and TepP and TARP was IP as above with anti-TARP or anti-TepP antibodies followed by immunoblot analysis with anti-TARP, -TepP and p-Tyr antibodies (E). Note relative delay in TepP tyrosine phosphorylation with respect to TARP. The prominent higher molecular weight tyrosine-phosphorylated band corresponds to TARP, as assessed by 2-color immunofluorescence-detection. In contrast, TepP only partially overlaps with the major 65–70 kDa tyrosine-phosphorylated proteins (F). MOMP: major outer membrane protein, Uninf: uninfected.</p
[Report to Chief J. E. Curry, by an unknown author #1]
No full textReport to Chief J. E. Curry, by an unknown author. The report contains a list of officers who gave depositions to the United States Attorney
[Report to Chief J. E. Curry, by an unknown author #2]
No full textReport to Chief J. E. Curry, by an unknown author. The report contains a list of officers who gave depositions to the United States Attorney
Global transcriptional profiling links TepP function to immune-related responses.
No full text<p><b>A</b>) Microarray analysis identified 33 host genes that have >1.5 fold-change in transcription between recombinant TepP<sup>W103*</sup> strains (CTL2-M062G1) transformed with empty vector (Vec) and vector containing wild type <i>tepP</i> gene (pTepP). A2EN epithelial cells were infected for 4 hours. Results shown were from duplicate biological samples. <b>B</b>) Q-PCR results validated transcriptional change observed for 5 immune-related genes at 4 hpi. Fold-change in mRNA abundance was normalized to CTL2-M062G1 (Vec). <b>C</b>) TepP-dependent elevation of IFIT genes expression at 8 hpi. Q-PCR results of MAP3k8, IFIT1 and IFIT2 mRNA in A2EN cells infected as in (A). All data shown were as means ± standard deviations from three independent biological replicates. *, <i>P</i><0.05; **, <i>P</i><0.01; ***, <i>P</i><0.001 (one-way analysis of variance and Tukey's multiple-comparison test).</p
TepP is required for the recruitment of Crk to nascent inclusions.
No full text<p><b>A</b>) Physical map of single nucleotide variants identified in strain CTL2-M062, a chemically derived LGV-L2 mutant containing a nonsense mutation in the codon for amino acid 103 of TepP. Non synonymous mutations were identified by whole genome sequencing and verified by Sanger sequencing. Key differences from the parental reference strain are highlighted. <b>B–C</b>) TepP is required for the accumulation of major tyrosine-phosphorylated proteins in <i>Chlamydia</i> infected cells. Immunoblot analysis of the EB lysate of CTL2-M062 shows no detectable levels of TepP compared to wild type LGV-L2 (WT) (B). HeLa cells were infected with wild type LGV-L2 (WT) or CTL2-M062 at an MOI of 50 and samples were collected at indicated time points and analyzed by immunoblot with antibodies against p-Tyr and MOMP. Arrows indicate major TepP-dependent tyrosine-phosphorylated proteins (C). <b>D</b>) Crk does not associate with nascent inclusions in CTL2-M062 infected cells. Immunofluorescence staining of HeLa cells infected with wild type LGV-L2 and CTL2-M062 for 8 h. Cells were stained with anti-Crk (red) and anti-MOMP (green) antibodies, and with DAPI (blue). Crk recruitment was absent in a TepP deficient strain. Scale bar: 20 µm. <b>E–F</b>) The <i>tepP</i> mutation is genetically linked to the lack of the infection-induced p65-70 kDa tyrosine-phosphorylated proteins and the loss of Crk recruitment to nascent inclusions. Various recombinant strains generated from a cross between CTL2-M062 and wild type LGV-L2 were tested for TepP expression (<a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1003954#ppat.1003954.s004" target="_blank">Fig. S4</a>, <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1003954#ppat.1003954.s005" target="_blank">S5</a>) and tested for the presence of the p65-70 kDa p-Tyr proteins (E) and recruitment of Crk to nascent inclusions (F).</p
Crk-I and Crk-II bind to tyrosine-phosphorylated TepP and are recruited to nascent inclusions early during infection.
No full text<p><b>A</b>) Cartoon schematic of TepP phosphorylation sites identified by mass spectrometry (<a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1003954#ppat.1003954.s001" target="_blank">Fig. S1</a>). <b>B</b>) CrkI and CrkII co-IP with TepP. HeLa cells were infected with L2 at MOI of 100 for 1, 2, 4, 6 and 8 hours. Cell lysates were immunoprecipitated with anti-TepP antibodies and the bound proteins were analyzed by immunoblot with anti-Crk, TepP and p-Tyr antibodies. Both isoforms of Crk, CrkI (lower band) and CrkII (upper band), co-IP with TepP. Uninf-uninfected. <b>C</b>) CrkI-II is recruited to <i>Chlamydia</i> entry sites and nascent inclusions. HeLa cells were infected with L2 at an MOI of 20 for 1 and 8 hours. Cells were fixed and stained with anti-MOMP (green) and anti-Crk (red) antibodies. Left panels are magnified images from boxed areas. Examples of sites of association between bacteria and Crk are marked by arrows. DAPI was used to stain nucleic acids. CrkI-II was recruited to <i>Chlamydia</i> entry sites by 1 hpi and this association continued after the nascent inclusions had trafficked to the host cells' perinuclear region.</p
Genetic complementation of a <i>Chlamydia tepP</i> mutant restores the normal tyrosine-phosphorylation pattern of multiple proteins and rescues Crk recruitment to nascent inclusions.
No full text<p><b>A</b>) The recombinant TepP<sup>W103*</sup> strain CTL2-M062G1 was transformed with an empty vector (Vec.) or a vector harboring the wild type <i>tepP</i> gene (pTepP). Immunoblot analysis of EBs derived from these strains with anti-TepP antibodies confirms the complementation of TepP expression to wild type levels (LGV-L2). Slc1 and RpoB/B' levels are shown as loading controls. <b>B</b>) The complemented recombinant TepP<sup>W103*</sup> strain restored the pattern of tyrosine-phosphorylation induced during <i>Chlamydia</i> infection. Confluent HeLa cells were infected with CTL2-M062G1 strains described in (A) at an MOI of 50 and total protein lysates were collected at indicated time points. Samples were subjected to immunoblot analysis with antibodies against p-Tyr and MOMP. Arrows indicate phosphotyrosine bands restored after TepP complementation. <b>C</b>) The complemented recombinant TepP<sup>W103*</sup> strains rescued Crk recruitment to nascent <i>Chlamydia</i> inclusions. Cells were infected for 8 hours with CTL2-M062G1 strains described in (A) at an MOI of 20, and immunostained with anti-Crk (red) and anti-MOMP (green) antibodies, and DAPI (blue).</p
Murder on the mountain: author talk with Peter J. Wosh
No full textAuthor talk by Peter J. Wosh on May 5th, 2022, on his book, "Murder on the Mountain: crime, passion, and punishment in gilded age New Jersey.
Mr. Melvin J. Collier, RWWL AUC, June 2011
No full textThis video is a conversation with Mr. Melvin J. Collier. Mr. Collier talks about his book, "From Mississippi to Africa: A Journey of Discovery". Daniel Le, AUC Woodruff Library, is the interviewer
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