Living Mercy: Reflecting on the Vocation and Values of Salve Regina University

With this collection of essays, we honor the vocation and spirit of mercy that has enlivened and guided Salve Regina University for the last 75 years. Inspired by the accomplishments of the past and looking forward to the call of the future, these essays provide a starting point for University-wide conversations to support Salve Regina in discerning how it will move into the increasingly complex challenges of the future. Salve\u27s tradition of mercy is rooted in the example of Catherine McAuley, who founded the Sisters of Mercy in 19th-century Dublin, Ireland. It is a model of faith expressed through action and maintains that each person is called to and capable of contributing to the common good by responding to the needs of the day, to respond to the suffering and injustice of each era.Attending to this spirit of mercy that continues to guide our University, this project considers how the six core values of Salve’s Strategic Compass – purpose-driven education, respect and dignity for all, mercy community, integrity, faith and spirituality, and compassionate service and solidarity – relate to our shared mercy, Catholic heritage, and the mercy vocational paradigm. Exploring how to re-root and re-frame these values, we approached the project as a vocationally oriented narrative. This type of narrative focuses on the call and vocation, as well as the patterns of meaning that shape the unique identity of an institution in its founding and how the institution has evolved and changed in response to the claims and context of social and historic dynamics. Thus, these six essays are harmonized by a three-fold critical-creative structure that attends to the dynamic experience of the call and spirit of mercy modeled in the founding of the University, how we presently live this call, and envision the challenges and possibilities that lie on the horizon. We employed the perspectives of Foundations, Living Presence, and Horizons to frame an analogical exploration of the unique character, actions and ideals that have inspired and sustained the vocation and mission of Salve Regina University, and may be creatively transferred to shaping the horizon for future generations of students.Celebrating the 75th anniversary of the founding of Salve Regina University, we invite readers to reflect on this collection of essays and then to join the conversations that are to follow as we continue to discern the path forward as Salve takes its next steps into the future.https://digitalcommons.salve.edu/fac_staff_ebooks/1006/thumbnail.jp

Long-Lasting Effects of Sepsis on Circadian Rhythms in the Mouse

Daily patterns of activity and physiology are termed circadian rhythms and are driven primarily by an endogenous biological timekeeping system, with the master clock located in the suprachiasmatic nucleus. Previous studies have indicated reciprocal relationships between the circadian and the immune systems, although to date there have been only limited explorations of the long-term modulation of the circadian system by immune challenge, and it is to this question that we addressed ourselves in the current study. Sepsis was induced by peripheral treatment with lipopolysaccharide (5 mg/kg) and circadian rhythms were monitored following recovery. The basic parameters of circadian rhythmicity (free-running period and rhythm amplitude, entrainment to a light/dark cycle) were unaltered in post-septic animals compared to controls. Animals previously treated with LPS showed accelerated re-entrainment to a 6 hour advance of the light/dark cycle, and showed larger phase advances induced by photic stimulation in the late night phase. Photic induction of the immediate early genes c-FOS, EGR-1 and ARC was not altered, and neither was phase-shifting in response to treatment with the 5-HT-1a/7 agonist 8-OH-DPAT. Circadian expression of the clock gene product PER2 was altered in the suprachiasmatic nucleus of post-septic animals, and PER1 and PER2 expression patterns were altered also in the hippocampus. Examination of the suprachiasmatic nucleus 3 months after treatment with LPS showed persistent upregulation of the microglial markers CD-11b and F4/80, but no changes in the expression of various neuropeptides, cytokines, and intracellular signallers. The effects of sepsis on circadian rhythms does not seem to be driven by cell death, as 24 hours after LPS treatment there was no evidence for apoptosis in the suprachiasmatic nucleus as judged by TUNEL and cleaved-caspase 3 staining. Overall these data provide novel insight into how septic shock exerts chronic effects on the mammalian circadian system

Long-Lasting Effects of Sepsis on Circadian Rhythms in the Mouse

Daily patterns of activity and physiology are termed circadian rhythms and are driven primarily by an endogenous biological timekeeping system, with the master clock located in the suprachiasmatic nucleus. Previous studies have indicated reciprocal relationships between the circadian and the immune systems, although to date there have been only limited explorations of the long-term modulation of the circadian system by immune challenge, and it is to this question that we addressed ourselves in the current study. Sepsis was induced by peripheral treatment with lipopolysaccharide (5 mg/kg) and circadian rhythms were monitored following recovery. The basic parameters of circadian rhythmicity (free-running period and rhythm amplitude, entrainment to a light/dark cycle) were unaltered in post-septic animals compared to controls. Animals previously treated with LPS showed accelerated re-entrainment to a 6 hour advance of the light/dark cycle, and showed larger phase advances induced by photic stimulation in the late night phase. Photic induction of the immediate early genes c-FOS, EGR-1 and ARC was not altered, and neither was phase-shifting in response to treatment with the 5-HT-1a/7 agonist 8-OH-DPAT. Circadian expression of the clock gene product PER2 was altered in the suprachiasmatic nucleus of post-septic animals, and PER1 and PER2 expression patterns were altered also in the hippocampus. Examination of the suprachiasmatic nucleus 3 months after treatment with LPS showed persistent upregulation of the microglial markers CD-11b and F4/80, but no changes in the expression of various neuropeptides, cytokines, and intracellular signallers. The effects of sepsis on circadian rhythms does not seem to be driven by cell death, as 24 hours after LPS treatment there was no evidence for apoptosis in the suprachiasmatic nucleus as judged by TUNEL and cleaved-caspase 3 staining. Overall these data provide novel insight into how septic shock exerts chronic effects on the mammalian circadian system

Characterization of Metabolically Quiescent <i>Leishmania</i> Parasites in Murine Lesions Using Heavy Water Labeling

<div><p>Information on the growth rate and metabolism of microbial pathogens that cause long-term chronic infections is limited, reflecting the absence of suitable tools for measuring these parameters <i>in vivo</i>. Here, we have measured the replication and physiological state of <i>Leishmania mexicana</i> parasites in murine inflammatory lesions using ²H₂O labeling. Infected BALB/c mice were labeled with ²H₂O for up to 4 months, and the turnover of parasite DNA, RNA, protein and membrane lipids estimated from the rate of deuterium enrichment in constituent pentose sugars, amino acids, and fatty acids, respectively. We show that the replication rate of parasite stages in these tissues is very slow (doubling time of ~12 days), but remarkably constant throughout lesion development. Lesion parasites also exhibit markedly lower rates of RNA synthesis, protein turnover and membrane lipid synthesis than parasite stages isolated from <i>ex vivo</i> infected macrophages or cultured <i>in vitro</i>, suggesting that formation of lesions induces parasites to enter a semi-quiescent physiological state. Significantly, the determined parasite growth rate accounts for the overall increase in parasite burden indicating that parasite death and turnover of infected host cells in these lesions is minimal. We propose that the <i>Leishmania</i> response to lesion formation is an important adaptive strategy that minimizes macrophage activation, providing a permissive environment that supports progressive expansion of parasite burden. This labeling approach can be used to measure the dynamics of other host-microbe interactions <i>in situ</i>.</p></div

Stage-specific changes in <i>Leishmania</i> growth rates.

<p><b>A</b>. Schematic overview of ²H₂O labeling protocol. Parasite stages were cultivated axenically in the presence 5% ²H₂O, or isolated from infected macrophages or BALB/c lesion incubated or infused with ²H₂O (final concentration 5%). Parasite stages were harvested at multiple time points and extracts containing total DNA/RNA, or total proteins and lipids generated from purified parasite fraction. Levels of deuterium enrichment in constituent dRib/Rib, amino acids and fatty acids were subsequently quantitated by GC-MS. <b>B</b>. Kinetics of ²H-labeling of DNA dRib in cultured promastigotes (Pro^log, Pro^stat) and amastigotes (Ama^axenic), and in amastigotes isolated from macrophages (Ama^Mø) and murine lesions (Ama^lesion). The fraction of new cells (Y-axis) was calculated from the level of ²H-enrichment in dRib relative to maximum labeling observed in each parasite stage after long term (equilibrium) labeling. Calculated doubling times for each stage are shown in inset boxes. <b>C</b>. Comparative growth rates of different <i>Leishmania</i> stages, calculated from ²H-enrichment in dRib. <b>D</b>. Section of stained cutaneous lesion (with detail in insert) and calculated range of parasite numbers/phagolysosome. Abbreviations: dRib; deoxyribose, Rib; ribose.</p

Rates of protein turnover in cultured and intracellular <i>Leishmania</i> stages.

<p>Parasite stages were ²H₂O-labeled in culture or <i>in situ</i> in infected BALB/c mice and harvested at the indicated time points. Kinetics of ²H-labeling of proteinogenic alanine in (<b>A</b>) cultured parasite stages (Pro^log, Pro^stat, Ama^axenic) and (<b>B</b>) amastigotes isolated from BALB/c lesion (Ama^lesion). The fraction of new molecules (Y-axis) was calculated from the level of ²H-enrichment in alanine relative to maximum labeling observed in each parasite stage after long term labeling. Inset boxes in A and B show turnover (t_1/2) in days. <b>C</b>. Comparative rates of protein turnover in different <i>Leishmania</i> developmental stages. Note that similar estimates of protein turnover were obtained by measuring deuterium incorporation into other proteinogenic amino acids (<a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1004683#ppat.1004683.s006" target="_blank">S6 Fig</a>.).</p

Rates of RNA turnover in cultured and intracellular <i>Leishmania</i> stages.

<p>Kinetics of ²H-labeling of RNA ribose in (<b>A</b>) cultured parasite stages (Pro^log, Pro^stat, Ama^axenic) (<b>B</b>) amastigotes isolated from infected J774 macrophages (Ama^Mø) and (<b>C</b>) amastigotes isolated from BALB/c lesions (Ama^lesion). The fraction of new molecules (Y-axis) was calculated from the level of ²H-enrichment in Rib relative to maximum labeling observed in each parasite stage after long term labeling. Inset boxes shows estimated RNA turnover (t_1/2 in days) in each stage. <b>D</b>. Comparative rates of RNA turnover in different <i>Leishmania</i> developmental stages.</p

Lesion amastigotes utilize both salvage and <i>de novo</i> biosynthetic pathways to supply their fatty acid needs.

<p><b>A</b>. The maximum level of ²H-enrichment (EM1, %) in major cellular fatty acids of Pro^log and Ama^lesion were determined after labeling for 7 days and >6 weeks, respectively. ²H-enrichment in the total plasma lipid of infected mice was also measured to determine the potential contribution of labeled host fatty acids to the parasite labeling. Note that while saturated and unsaturated C18 fatty acids are predominant fatty acids in both stages, the fatty acid composition of Ama^lesion differs from cultured promastigotes in containing elevated levels of C20:4 n-6, and polyunsaturated very long chain fatty acids (C22:4 n-6, C22:6 n-3) (<a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1004683#ppat.1004683.s008" target="_blank">S8 Fig</a>.). The C:D nomenclature refers to overall chain length and number of double bonds in each fatty acid, respectively. n-3 and n-6 refers to the two major biosynthetic pathways involved in unsaturated fatty acid biosynthesis (where-3 and-6 refer to the position of double bond relative to the methyl carbon). <b>B</b>. Stage-specific differences in the levels of ²H-enrichment in fatty acid pools can be used to infer the contributions of <i>de novo</i> biosynthesis and salvage pathways. In particular, levels of ²H-enrichment in the major Ama^lesion C18 fatty acids, C18:1 (oleic acid) and C18:2 (linoleic acid), were appreciably higher than in host plasma, indicating that these stages are dependent on <i>de novo</i> biosynthesis. Conversely, the elevated levels of ²H-enrichment in C20:4 n-6 and C22:6n-3 compared to C18:1 precursor (comparable to plasma pools) indicate that these very long chain polyunsaturated fatty acids are primarily scavenged from the host cell.</p

Influence of sire fertility status on conceptus-induced transcriptomic response of the bovine endometrium

The aim was to examine the effect of sire fertility status on conceptus-induced changes in the bovine endometrial transcriptome. To generate elongated conceptuses, Day 7 blastocysts produced in vitro using frozen-thawed sperm from Holstein Friesian bulls (3 High fertility, HF and 3 Low fertility, LF) were transferred in groups of 5–10 into synchronized heifers (n = 7 heifers per bull) and recovered following slaughter on Day 15. Day 15 endometrial explants recovered from the uterine horn ipsilateral to the corpus luteum were recovered from synchronized cyclic heifers (n = 4). Explants from each heifer were co-cultured for 6 h in RPMI medium alone (Control) or with 100 ng/ml ovine recombinant interferon tau (IFNT) or with a single conceptus from each HF or LF bull. After 6 h, explants were snap frozen and stored at −80°C. Extracted mRNA was subjected to RNA-seq and the resulting data were analyzed with R software. The numbers of differentially expressed genes (DEG; FDR<0.05) were: HF vs. Control: 956; LF vs. Control: 1021; IFNT vs. Control: 1301; HF vs. LF: 2. Unsurprisingly, the majority of DEG (658) were common to all comparisons and were related to IFNT-induced changes in the endometrium. Prior to applying the adjusted p-value, there were 700 DEG between HF and LF, with 191 and 509 genes more expressed in HF or LF, respectively (p < 0.05). Overrepresentation analysis of KEGG pathways (FDR<0.05), revealed that DEG with higher expression in LF were involved in cell cycle and proteolysis, while those upregulated DEG by HF conceptuses were strongly associated with immune process pathways, such as TNF, NF-kappa B, cytokine-cytokine receptor interaction, and TLR signaling.These pathways were also enriched by DEG upregulated by IFNT compared to the Control. Furthermore, only the HF, and not the LF group, affected the expression of most genes in these pathways (p < 0.05) according to a negative binomial regression model. Finally, a weighted gene co-expression network analysis revealed two clusters of co-expressed genes associated with the HF conceptuses (p < 0.05), which were also enriched for the aforementioned pathways. In conclusion, HF conceptuses, similar to IFNT treatment, stimulated multiple pathways involved in immune response, which were apparently not affected by LF conceptuses.</p