Understanding Meroitic Pottery and Its Production – Research Design and Methodology of an Interdisciplinary Research Project

This paper describes a project investigating the archaeological remains of a pottery workshop and the associated ceramics at Musawwarat es-Sufra, an archaeological site of the Meroitic period (c. 300 BC–350 AD) in Sudan. It outlines the trajectory of a complex research approach, integrating archaeological, ceramological, archaeometric, ethnoarchaeological and geophysical components in order to make maximum use of the unique potential of the site. By discussing the setup and the trajectory of the project and how the results – expected and unexpected – of the individual components informed subsequent steps and the progress of the overall research strategy, the authors offer a contribution to the methodology of integrated research in the field of archaeological ceramic studies

E-cigarette exposure delays implantation and causes reduced weight gain in female offspring exposed in utero

Electronic nicotine delivery system (e-cigarette) use is prevalent among pregnant women as a seemingly safe alternative to traditional tobacco use, known to result in fetal developmental abnormalities and impaired fertility of male offspring. However, little is known about the effects of e-cigarette use on fertility or pregnancy outcomes. A successful pregnancy is initiated by a multitude of dynamic molecular alterations in the uterus resulting in embryo implantation at day 4.5 in the mouse. We examined whether e-cigarette exposure impairs implantation and offspring health. Pregnant C57BL/6J mice were exposed five times a week to e-cigarette vapor or sham. After 4 months, e-cigarette exposed dams exhibited a significant delay in the onset of the first litter. Furthermore, exposure of new dams in early pregnancy significantly impaired embryo implantation, as evidenced by nearly complete absence of implantation sites in e-cigarette–exposed animals at day 5.5, despite exhibiting high levels of progesterone, an indicator of pregnancy. RNA microarray from day 4.5 pseudopregnant mice revealed significant changes in the integrin, chemokine, and JAK signaling pathways. Moreover, female offspring exposed to e-cigarettes in utero exhibited a significant weight reduction at 8.5 months, whereas males exhibited a slight but nonsignificant deficiency in fertility. Thus, e-cigarette exposure in mice impairs pregnancy initiation and fetal health, suggesting that e-cigarette use by reproductive-aged women or during pregnancy should be considered with caution

FOXO1 regulates uterine epithelial integrity and progesterone receptor expression critical for embryo implantation.

Successful embryo implantation requires a receptive endometrium. Poor uterine receptivity can account for implantation failure in women who experience recurrent pregnancy loss or multiple rounds of unsuccessful in vitro fertilization cycles. Here, we demonstrate that the transcription factor Forkhead Box O1 (FOXO1) is a critical regulator of endometrial receptivity in vivo. Uterine ablation of Foxo1 using the progesterone receptor Cre (PgrCre) mouse model resulted in infertility due to altered epithelial cell polarity and apoptosis, preventing the embryo from penetrating the luminal epithelium. Analysis of the uterine transcriptome after Foxo1 ablation identified alterations in gene expression for transcripts involved in the activation of cell invasion, molecular transport, apoptosis, β-catenin (CTNNB1) signaling pathway, and an increase in PGR signaling. The increase of PGR signaling was due to PGR expression being retained in the uterine epithelium during the window of receptivity. Constitutive expression of epithelial PGR during this receptive period inhibited expression of FOXO1 in the nucleus of the uterine epithelium. The reciprocal expression of PGR and FOXO1 was conserved in human endometrial samples during the proliferative and secretory phase. This demonstrates that expression of FOXO1 and the loss of PGR during the window of receptivity are interrelated and critical for embryo implantation

<i>Egfr</i> ablation causes blastocyst implantation site demise.

<p>(<b>A–F</b>) Gross morphology of pregnant uteri. (<b>A–C</b>) <i>Egfr^f/f</i> and (<b>D–F</b>) <i>Egfr^d/d</i> females were mated with wild-type males. The morning observance of a vaginal plug was considered day 0.5 of pregnancy. Pregnancy was assessed at (<b>A,D</b>) d5.5, (<b>B,E</b>) d6.5 and (<b>C,F</b>) d9.5. Scale bars: 1 cm. (<b>G</b>) Average number of implantation sites observed. (<b>H</b>) Average lateral diameter of implantation sites. Numbers represent average +/− the SEM. **-<i>p</i><0.01; ***-<i>p</i><0.001.</p

Ablation of <i>Egfr</i> does not impair acute hormone responsiveness.

<p>Mice were ovariectomized and treated with: (<b>A,B</b>) estrogen for 3 d or (<b>C</b>) progesterone for 6 h. (<b>A</b>) Measurement of the induction of uterine wet weight in response to estrogen. qPCR measurement of hormone target genes after (<b>B</b>) 3 d of estrogen or (<b>C</b>) 6 h of progesterone. Average +/− SEM. *-<i>p</i><0.05; **-<i>p</i><0.01; ***-<i>p</i><0.001.</p

<i>Egfr</i> ablation results in major defects in decidualization.

<p>Ovariectomized mice were treated with exogenous hormones and a deciduogenic stimulus was administered to one uterine horn. Gross uterine morphology of uteri (<b>A</b>) 1 day, (<b>B</b>) 2 days, or (<b>C</b>) 5 days after deciduogenic stimulus. Scale bar: 1 cm. (<b>D</b>) Wet weight ratio of stimulated uterine horn relative to unstimulated horn. Histological analysis of day 2 stimulated uterine cross-sections comparing (<b>E–G</b>) <i>Egfr^f/f</i> and (<b>H–J</b>) <i>Egfr^d/d</i>. (<b>E,H</b>) Proliferative marker phospho-histone H3, (<b>F,I</b>) apoptosis via TUNEL assay, and (<b>G,J</b>) differentiation via alkaline phosphatase staining. Scale bars: 100 µm (E,H,F,I) and 50 µm (G,J). (<b>K</b>) Measurement of the induction of mRNA expression of known decidual regulators on day 1 uteri by quantitative real-time PCR. Average +/− SEM. *-<i>p</i><0.05; **-<i>p</i><0.01; ***-<i>p</i><0.001. n.s. - not significant.</p

Direct and decidual-dependent kinome alterations following EGFR attenuation in HESC by kinase antibody array.

<p>(<b>A–C</b>) HESC were transfected with non-targeting or <i>EGFR</i> siRNA and treated with (<b>A</b>) HB-EGF for 30 mins or (<b>B</b>) deciduogenic hormones for 72 h to determine the effect on 44 different kinase substrates via kinase antibody arrays. (<b>C</b>) Relative (si<i>EGFR</i> vs non-targeting siRNA) quantification of antibodies exhibiting an absolute fold change >1.5. (T) indicates an antibody recognizing total proteins; all others are phosphorylation specific. (<b>D</b>) Validation of kinase array results by western blot using independent samples. (<b>E</b>) HESC were transfected with non-targeting (NT), <i>WNK1</i>, or <i>AKT1S1</i> siRNA, treated with deciduogenic hormones for 6 d and the expression of decidual markers was measured by qPCR. Numbers represent average +/− the SEM. **-p<0.01; ***-p<0.001.</p