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Primer information.

This lists the forward (FWD) and reverse (RVS) primers that were used for PCR of each target gene listed using zebrafish cDNA. (TIF)</p

Zebrafish expression of Tier 2 pharmacogenes.

A. Previously published literature housing information on expression of Tier 2 pharmacogenes in zebrafish. B. Tier 2 pharmacogene expression in zebrafish: “Yes” indicates expression was noted, “No” indicates no expression found. C. RT-PCR gel results for each Tier 2 zebrafish ortholog not previously identified in the literature. Gene name is listed at the top with the 3-days post fertilization (dpf) and 5-dpf included under the respective gene in alphabetical order. Each gel had a 100-base pair (bp) ladder (left) and a negative control (- ctrl, left).</p

Gene ontology analysis for cellular components of tiered pharmacogenes.

A. A map with the Cellular Component terms listed (left), amino acid sequence conservation in 3 groups: > 70% = High (green to yellow, left to right), 70% > x > 55% = Middle (yellow to orange, left to right), and < 55% = Low (orange to red, left to right). The genes are placed within the respective horizontal location based on their % conservation. B. Average amino acid conservation for each Cellular Component term +/- SD. C. The number of pharmacogenes in each category (High = green, Middle = yellow, Low = red) for each Cellular Component term.</p

Pharmacogene orthologs and amino acid conservation.

Tables with Tier 1 (A) and Tier 2 (B) human pharmacogenes (left), the orthologous zebrafish gene (middle), and the conservation in amino acid sequence as determined by NCBI BLAST. C. Summary tables of averages, standard deviation (SD), minimum, and maximums for Tier 1, Tier 2, and combined. *indicates ZFIN/Alliance-indicated ortholog.</p

Comparison of tiered pharmacogene sources.

Venn diagrams of Tier 1 (A) and Tier 2 (B) pharmacogenes included from the three sources: PharmGKB (red), CPIC (green), and the FDA (blue).</p

Sequence alignment example.

Here we provide an example of the most similar (in terms of amino acid sequence) Tier 2 gene (HMGCR) to visualize the similarities amongst amino acids over the length of the sequence. (TIF)</p

Clarithromycin prevents preterm birth and neonatal mortality by dampening alarmin-induced maternal–fetal inflammation in mice

Abstract Background One of every four preterm neonates is born to a woman with sterile intra-amniotic inflammation (inflammatory process induced by alarmins); yet, this clinical condition still lacks treatment. Herein, we utilized an established murine model of sterile intra-amniotic inflammation induced by the alarmin high-mobility group box-1 (HMGB1) to evaluate whether treatment with clarithromycin prevents preterm birth and adverse neonatal outcomes by dampening maternal and fetal inflammatory responses. Methods Pregnant mice were intra-amniotically injected with HMGB1 under ultrasound guidance and treated with clarithromycin or vehicle control, and pregnancy and neonatal outcomes were recorded (n = 15 dams each). Additionally, amniotic fluid, placenta, uterine decidua, cervix, and fetal tissues were collected prior to preterm birth for determination of the inflammatory status (n = 7–8 dams each). Results Clarithromycin extended the gestational length, reduced the rate of preterm birth, and improved neonatal mortality induced by HMGB1. Clarithromycin prevented preterm birth by interfering with the common cascade of parturition as evidenced by dysregulated expression of contractility-associated proteins and inflammatory mediators in the intra-uterine tissues. Notably, clarithromycin improved neonatal survival by dampening inflammation in the placenta as well as in the fetal lung, intestine, liver, and spleen. Conclusions Clarithromycin prevents preterm birth and improves neonatal survival in an animal model of sterile intra-amniotic inflammation, demonstrating the potential utility of this macrolide for treating women with this clinical condition, which currently lacks a therapeutic intervention.http://deepblue.lib.umich.edu/bitstream/2027.42/173661/1/12884_2022_Article_4764.pd

Pregnancy imparts distinct systemic adaptive immune function

ProblemPregnancy represents a state of systemic immune activation that is primarily driven by alterations in circulating innate immune cells. Recent studies have suggested that cellular adaptive immune components, T cells and B cells, also undergo changes throughout gestation. However, the phenotypes and functions of such adaptive immune cells are poorly understood. Herein, we utilized high-dimensional flow cytometry and functional assays to characterize T-cell and B-cell responses in pregnant and non-pregnant women.MethodsPeripheral blood mononuclear cells from pregnant (n = 20) and non-pregnant (n = 25) women were used for phenotyping of T-cell and B-cell subsets. T-cell proliferation and B-cell activation were assessed by flow cytometry after in vitro stimulation, and lymphocyte cytotoxicity was evaluated by using a cell-based assay. Statistical comparisons were performed with linear mixed-effects models.ResultsPregnancy was associated with modestly enhanced basal activation of peripheral CD4+ T cells. Both CD4+ and CD8+ T cells from pregnant women showed increased activation-induced proliferation; yet, a reduced proportion of these cells expressed activation markers compared to non-pregnant women. There were no differences in peripheral lymphocyte cytotoxicity between study groups. A greater proportion of B cells from pregnant women displayed memory-like and activated phenotypes, and such cells exhibited higher activation following stimulation.ConclusionMaternal circulating T cells and B cells display distinct responses during pregnancy. The former may reflect the unique capacity of T cells to respond to potential threats without undergoing aberrant activation, thereby preventing systemic inflammatory responses that can lead to adverse perinatal consequences.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/175120/1/aji13606.pdfhttp://deepblue.lib.umich.edu/bitstream/2027.42/175120/2/aji13606_am.pd