Regulation of 20α-Hydroxysteroid Dehydrogenase Expression in Term Pregnant Human Myometrium Ex Vivo.

Metabolic inactivation of progesterone within uterine myocytes by 20α-hydroxysteroid dehydrogenase (20α-HSD) has been postulated as a mechanism contributing to functional progesterone withdrawal at term. In humans, 20α-HSD is encoded by the gene AKR1C1. Myometrial AKR1C1 mRNA abundance has been reported to increase significantly during labor at term. In spontaneous preterm labor, however, we previously found no increase in AKR1C1 mRNA level in the myometrium except for preterm labor associated with clinical chorioamnionitis. This suggests that increased 20α-HSD activity is a mechanism through which inflammation drives progesterone withdrawal in preterm labor. In this study, we have determined the effects of various treatments of therapeutic relevance on AKR1C1 expression in pregnant human myometrium in an ex vivo culture system. AKR1C1 expression increased spontaneously during 48 h culture (p < 0.0001), consistent with the myometrium transitioning to a labor-like phenotype ex vivo, as reported previously. Serum supplementation, prostaglandin F2α, phorbol myristate acetate, and mechanical stretch had no effect on the culture-induced increase, whereas progesterone (p = 0.0058) and cAMP (p = 0.0202) further upregulated AKR1C1 expression. In contrast, culture-induced upregulation of AKR1C1 expression was dose-dependently repressed by three histone/protein deacetylase inhibitors: trichostatin A at 5 (p = 0.0172) and 25 µM (p = 0.0115); suberoylanilide hydroxamic acid at 0.5 (p = 0.0070), 1 (p = 0.0045), 2.5 (p = 0.0181), 5 (p = 0.0066) and 25 µM (p = 0.0014); and suberoyl bis-hydroxamic acid at 5 (p = 0.0480) and 25 µM (p = 0.0238). We propose the inhibition of histone/protein deacetylation helps to maintain the anti-inflammatory, pro-quiescence signaling of progesterone in pregnant human myometrium by blocking its metabolic inactivation. Histone deacetylase inhibitors may represent a class of agents that preserve or restore the progesterone sensitivity of the pregnant uterus.

Clopidogrel Administration Impairs Post-Stroke Learning and Memory Recovery in Mice.

Clopidogrel, which is one of the most prescribed antiplatelet medications in the world, is given to stroke survivors for the prevention of secondary cardiovascular events. Clopidogrel exerts its antiplatelet activity via antagonism of the P2Y12 receptor (P2RY12). Although not widely known or considered during the initial clinical trials for clopidogrel, P2RY12 is also expressed on microglia, which are the brain's immune cells, where the receptor facilitates chemotactic migration toward sites of cellular damage. If microglial P2RY12 is blocked, microglia lose the ability to migrate to damaged sites and carry out essential repair processes. We aimed to investigate whether administering clopidogrel to mice post-stroke was associated with (i) impaired motor skills and cognitive recovery; (ii) physiological changes, such as survival rate and body weight; (iii) changes in the neurovascular unit, including blood vessels, microglia, and neurons; and (iv) changes in immune cells. Photothrombotic stroke (or sham surgery) was induced in adult male mice. From 24 h post-stroke, mice were treated daily for 14 days with either clopidogrel or a control. Cognitive performance (memory and learning) was assessed using a mouse touchscreen platform (paired associated learning task), while motor impairment was assessed using the cylinder task for paw asymmetry. On day 15, the mice were euthanized and their brains were collected for immunohistochemistry analysis. Clopidogrel administration significantly impaired learning and memory recovery, reduced mouse survival rates, and reduced body weight post-stroke. Furthermore, clopidogrel significantly increased vascular leakage, significantly increased the number and appearance of microglia, and significantly reduced the number of T cells within the peri-infarct region post-stroke. These data suggest that clopidogrel hampers cognitive performance post-stroke. This effect is potentially mediated by an increase in vascular permeability post-stroke, providing a pathway for clopidogrel to access the central nervous system, and thus, interfere in repair and recovery processes.

20α-Hydroxysteroid Dehydrogenase Expression in the Human Myometrium at Term and Preterm Birth: Relationships to Fetal Sex and Maternal Body Mass Index.

The mechanism by which human labor is initiated in the presence of elevated circulating progesterone levels remains unknown. Recent evidence indicates that the progesterone-metabolizing enzyme, 20α-hydroxysteroid dehydrogenase (20α-HSD), encoded by the gene AKR1C1, may contribute to functional progesterone withdrawal. We found that AKR1C1 expression significantly increased with labor onset in term myometrium, but not in preterm myometrium. Among preterm laboring deliveries, clinically diagnosed chorioamnionitis was associated with significantly elevated AKR1C1 expression. AKR1C1 expression positively correlated with BMI before labor and negatively correlated with BMI during labor. Analysis by fetal sex showed that AKR1C1 expression was significantly higher in women who delivered male babies compared to women who delivered female babies at term, but not preterm. Further, in pregnancies where the fetus was female, AKR1C1 expression positively correlated with the mother's age and BMI at the time of delivery. In conclusion, the increase in myometrial AKR1C1 expression with term labor is consistent with 20α-HSD playing a role in local progesterone metabolism to promote birth. Interestingly, this role appears to be specific to term pregnancies where the fetus is male. Upregulated AKR1C1 expression in the myometrium at preterm in-labor with clinical chorioamnionitis suggests that increased 20α-HSD activity is a mechanism through which inflammation drives progesterone withdrawal in preterm labor. The link between AKR1C1 expression and maternal BMI may provide insight into why maternal obesity is often associated with dysfunctional labor. Higher myometrial AKR1C1 expression in male pregnancies may indicate fetal sex-related differences in the mechanisms that precipitate labor onset at term.

Pre-gelled Electrode Grid for Self-Applied EEG Sleep Monitoring at Home.

The need for diagnostic capabilities for sleep disorders such as sleep apnea and insomnia far exceeds the capacity of inpatient sleep laboratories. Some home monitoring systems omit electroencephalography (EEG) because trained personnel may be needed to apply EEG sensors. Since EEG is essential for the detailed evaluation of sleep, better systems supporting the convenient and robust recording of sleep EEG at home are desirable. Recent advances in EEG acquisition with flex-printed sensors promise easier application of EEG sensor arrays for chronic recordings, yet these sensor arrays were not designed for sleep EEG. Here we explored the self-applicability of a new sleep EEG sensor array (trEEGrid) without prior training. We developed a prototype with pre-gelled neonatal ECG electrodes placed on a self-adhesive grid shape that guided the fast and correct positioning of a total of nine electrodes on the face and around the ear. Positioning of the sensors was based on the results of a previous ear-EEG sleep study (da Silva Souto et al., 2021), and included electrodes around the ear, one eye, and the chin. For comparison, EEG and electrooculogram channels placed according to the American Academy of Sleep Medicine criteria, as well as respiratory inductance plethysmography on thorax and abdomen, oxygen saturation, pulse and body position were included with a mobile polysomnography (PSG) system. Two studies with 32 individuals were conducted to compare the signal quality of the proposed flex-printed grid with PSG signals and to explore self-application of the new grid at home. Results indicate that the new array is self-applicable by healthy participants without on-site hands-on support. A comparison of the hypnogram annotations obtained from the data of both systems revealed an overall substantial agreement on a group level (Cohen's κ = 0.70 ± 0.01). These results suggest that flex-printed pre-gelled sensor arrays designed for sleep EEG acquisition can facilitate self-recording at home.

Flex-Printed Ear-EEG Sensors for Adequate Sleep Staging at Home.

A comfortable, discrete and robust recording of the sleep EEG signal at home is a desirable goal but has been difficult to achieve. We investigate how well flex-printed electrodes are suitable for sleep monitoring tasks in a smartphone-based home environment. The cEEGrid ear-EEG sensor has already been tested in the laboratory for measuring night sleep. Here, 10 participants slept at home and were equipped with a cEEGrid and a portable amplifier (mBrainTrain, Serbia). In addition, the EEG of Fpz, EOG_L and EOG_R was recorded. All signals were recorded wirelessly with a smartphone. On average, each participant provided data for M = 7.48 h. An expert sleep scorer created hypnograms and annotated grapho-elements according to AASM based on the EEG of Fpz, EOG_L and EOG_R twice, which served as the baseline agreement for further comparisons. The expert scorer also created hypnograms using bipolar channels based on combinations of cEEGrid channels only, and bipolar cEEGrid channels complemented by EOG channels. A comparison of the hypnograms based on frontal electrodes with the ones based on cEEGrid electrodes (κ = 0.67) and the ones based on cEEGrid complemented by EOG channels (κ = 0.75) both showed a substantial agreement, with the combination including EOG channels showing a significantly better outcome than the one without (p = 0.006). Moreover, signal excerpts of the conventional channels containing grapho-elements were correlated with those of the cEEGrid in order to determine the cEEGrid channel combination that optimally represents the annotated grapho-elements. The results show that the grapho-elements were well-represented by the front-facing electrode combinations. The correlation analysis of the grapho-elements resulted in an average correlation coefficient of 0.65 for the most suitable electrode configuration of the cEEGrid. The results confirm that sleep stages can be identified with electrodes placement around the ear. This opens up opportunities for miniaturized ear-EEG systems that may be self-applied by users.

ß3-Adrenergic receptor stimulation induces E-selectin-mediated adipose tissue inflammation.

Inflammation induced by wound healing or infection activates local vascular endothelial cells to mediate leukocyte rolling, adhesion, and extravasation by up-regulation of leukocyte adhesion molecules such as E-selectin and P-selectin. Obesity-associated adipose tissue inflammation has been suggested to cause insulin resistance, but weight loss and lipolysis also promote adipose tissue immune responses. While leukocyte-endothelial interactions are required for obesity-induced inflammation of adipose tissue, it is not known whether lipolysis-induced inflammation requires activation of endothelial cells. Here, we show that ß(3)-adrenergic receptor stimulation by CL 316,243 promotes adipose tissue neutrophil infiltration in wild type and P-selectin-null mice but not in E-selectin-null mice. Increased expression of adipose tissue cytokines IL-1ß, CCL2, and TNF-α in response to CL 316,243 administration is also dependent upon E-selectin but not P-selectin. In contrast, fasting increases adipose-resident macrophages but not neutrophils, and does not activate adipose-resident endothelium. Thus, two models of lipolysis-induced inflammation induce distinct immune cell populations within adipose tissue and exhibit distinct dependences on endothelial activation. Importantly, our results indicate that ß(3)-adrenergic stimulation acts through up-regulation of E-selectin in adipose tissue endothelial cells to induce neutrophil infiltration.

Synergistic proinflammatory responses induced by polymicrobial colonization of epithelial surfaces.

The epithelial surfaces of the upper respiratory tract are continuously exposed to a wide variety of commensal microorganisms. In addition to acting as a physical barrier, epithelial cells respond to specific microbial products with the generation of signals, such as cytokines, that trigger inflammation. Because they are common components of the nasopharyngeal flora that share the potential to cause disease, we investigated the effects of Haemophilus influenzae and Streptococcus pneumoniae, alone and in combination, on human respiratory epithelial cells in culture and in a murine model of nasopharyngeal colonization. Exposure of A549 or Detroit 562 epithelial cells to both S. pneumoniae and H. influenzae led to a synergistic increase in production of IL-8, the major neutrophil chemokine in the airway, through an NF-kappaB-dependent mechanism. Likewise, nasal cocolonization of mice caused a synergistic rise in local production of macrophage inflammatory protein 2 in nasal lavage fluid and subsequent recruitment of neutrophils. This synergistic effect depended on production of the pore-forming cytolytic toxin, pneumolysin, by S. pneumoniae and activation of host p38 mitogen-activated protein kinase. Although both H. influenzae and S. pneumoniae have ligands for Toll-like receptors (TLRs) TLR2 and TLR4, synergistic activation was TLR2- and TLR4-independent. Thus, epithelial surfaces are capable of amplifying proinflammatory responses during concurrent stimulation by multiple microbial species. These synergistic responses, demonstrated both in vitro and in vivo, may contribute to inflammation of heavily colonized mucosal barriers.