How stands our press?

https://stars.library.ucf.edu/prism/1707/thumbnail.jp

Devotions for Lent 2023 Hymns of Lent

This Lent, we will continue reflecting on hymns of faith, namely, some of our most beloved Lenten hymns. 10 such hymns have been chosen to fill the 40(+) days of Lent. Therefore, this devotional, different from previous editions, does not proceed on a weekly basis, but merely flows from one hymn to the next. Also different from previous editions, the devotional reflections are specifically based on the stanzas of the selected hymns. Therefore, each day’s reflection features the text of the hymn stanza, a devotion based on that stanza, a prayer, and then a Scripture passage or passages for further meditation. I pray these reflections may be of edification for you during this Lenten season.https://scholar.csl.edu/osp/1022/thumbnail.jp

Writing in Britain and Ireland, c. 400 to c. 800

No abstract available

Die Press loday? S?l. n?. n?1930>pet?8>95 p? (Reprinted from THE JHATTO).

LEIDSSTELSELOPLADEN-RUG0

Germany embattled; an American interpretation,

Mode of access: Internet

A Forward Genetic Screen Reveals that Calcium-dependent Protein Kinase 3 Regulates Egress in <em>Toxoplasma</em>

<div><p>Egress from the host cell is a crucial and highly regulated step in the biology of the obligate intracellular parasite, <em>Toxoplasma gondii</em>. Active egress depends on calcium fluxes and appears to be a crucial step in escaping the attack from the immune system and, potentially, in enabling the parasites to shuttle into appropriate cells for entry into the brain of the host. Previous genetic screens have yielded mutants defective in both ionophore-induced egress and ionophore-induced death. Using whole genome sequencing of one mutant and subsequent analysis of all mutants from these screens, we find that, remarkably, four independent mutants harbor a mis-sense mutation in the same gene, <em>TgCDPK3</em>, encoding a calcium-dependent protein kinase. All four mutations are predicted to alter key regions of TgCDPK3 and this is confirmed by biochemical studies of recombinant forms of each. By complementation we confirm a crucial role for TgCDPK3 in the rapid induction of parasite egress and we establish that TgCDPK3 is critical for formation of latent stages in the brains of mice. Genetic knockout of TgCDPK3 confirms a crucial role for this kinase in parasite egress and a non-essential role for it in the lytic cycle.</p> </div

Phenotypic complementation of MBE1.1.

<p>A. Intracellular parasites of the parental (RHΔ<i>hpt</i>), mutant (MBE1.1), complemented (MBE1.1+sagCDPK3::HA) strains, as well as MBE1.1 parasites complemented with TgCDPK3 carrying the T239I mutation (MBE1.1+sagCDPK(T239I)::HA) were exposed to 1 µM A23187 for the time points indicated in the graph. Percentage egress represents the number of lysed vacuoles divided by the total number of vacuoles (lysed+intact). B. Exogenous copy of CDPK3::HA in the strains complemented with the wild type (MBE1.1+sagCDPK3::HA) or mutant gene (MBE1.1+sagCDPK3(T239I)::HA) was detected by western blot analysis using HA antibodies. Antibodies against the surface protein Sag1 were used to show equal loading. C. Extracellular parasites of the indicated strains were exposed to 1 µM A23187 for 45 minutes before being added to cells. Percentage survival was calculated by dividing the number of plaques formed by parasites treated with A23187 divided by the number of plaques formed by untreated parasites. For each treatment, all plaques in a well of a 12-well plate were counted. In B and C, each data point represents the average of four independent experiments and the error bars represent the standard deviation.</p

Establishment and analysis of a <i>Tgcdpk3</i> knockout strain.

<p>A. Schematic diagram of the TgCDK3 genomic locus and the region disrupted in the knockout strain. The top black line represents the genomic DNA region with each tick being 1,000 bases. The gray box represents the region of the TgCDPK3 locus that is replaced by the <i>hpt</i> selectable marker (dhfrHXGPRTdhfr), which is transcribed in the opposite direction from TgCDPK3 (as indicated by arrow pointing left). The relative positions of exons (white boxes) and introns (dashed lines) are shown below. Black arrowhead indicates position of start codon while the white arrowhead points at the stop codon. Lines labeled a, b, c and d represent the regions amplified by PCR to test for the disruption of <i>TgCDPK3</i>. B. PCR products of the parental strain, RHΔ<i>hpt</i>Δku80, and the TgCDPK3 KO strain, RHΔ<i>hpt</i>Δ<i>ku80</i>Δ<i>cdpk3</i>+HPT are shown. Labels above lanes indicate the DNA fragment amplified. Fragments a, b and c cannot be amplified if the T<i>gCDPK3</i> locus has been disrupted by the <i>hpt</i> marker. Fragment d is expanded from both the parental and knockout strain and it serves as a control. C. Intracellular parasites of either the parental or knockout strain were treated with 1 µM A23187 for the time indicated. Percentage egress represents the number of lysed vacuoles divided by the total number of vacuoles (lysed+intact). Each data point represents the average of four independent experiments and the error bars represent the standard deviation. D. Extracellular parasites of either the parental or the knockout strain were exposed to 1 µM A23187 for 45 minutes before being added to cells. Percentage survival was calculated by dividing the number of plaques formed by parasites treated with A23187 divided by the number of plaques formed by untreated parasites. Each data point represents the average of three independent experiments and the error bars represent the standard deviation. E. Parasites were allowed to settle on glass in intracellular buffer, which was changed to extracellular buffer to initiate motility. After 15 minutes parasites were fixed and the percentage of parasites that had moved was determined by counting parasites that were associated with trails of surface proteins left behind during gliding. Data is from 3 independent studies and the error bars are the standard deviation.</p