Synergistic Pulmonoprotective Effect of Natural Prolyl Oligopeptidase Inhibitors in In Vitro and In Vivo Models of Acute Respiratory Distress Syndrome.

Acute respiratory distress syndrome (ARDS) is a highly morbid inflammatory lung disease with limited pharmacological interventions. The present study aims to evaluate and compare the potential pulmonoprotective effects of natural prolyl oligopeptidase (POP) inhibitors namely rosmarinic acid (RA), chicoric acid (CA), epigallocatechin-3-gallate (EGCG) and gallic acid (GA), against lipopolysaccharide (LPS)-induced ARDS. Cell viability and expression of pro-inflammatory mediators were measured in RAW264.7 cells and in primary murine lung epithelial and bone marrow cells. Nitric oxide (NO) production was also assessed in unstimulated and LPS-stimulated RAW264.7 cells. For subsequent in vivo experiments, the two natural products (NPs) with the most favorable effects, RA and GA, were selected. Protein, cell content and lipid peroxidation levels in bronchoalveolar lavage fluid (BALF), as well as histopathological changes and respiratory parameters were evaluated in LPS-challenged mice. Expression of key mediators involved in ARDS pathophysiology was detected by Western blotting. RA and GA favorably reduced gene expression of pro-inflammatory mediators in vitro, while GA decreased NO production in macrophages. In LPS-challenged mice, RA and GA co-administration improved respiratory parameters, reduced cell and protein content and malondialdehyde (MDA) levels in BALF, decreased vascular cell adhesion molecule-1 (VCAM-1) and the inducible nitric oxide synthase (iNOS) protein expression, activated anti-apoptotic mechanisms and down-regulated POP in the lung. Conclusively, these synergistic pulmonoprotective effects of RA and GA co-administration could render them a promising prophylactic/therapeutic pharmacological intervention against ARDS.

Cardioprotective potential of oleuropein, hydroxytyrosol, oleocanthal and their combination: Unravelling complementary effects on acute myocardial infarction and metabolic syndrome.

Clinical studies have previously established the role of olive products in cardiovascular disease (CVD) prevention, whilst the identification of the responsible constituents for the beneficial effects is still pending. We sought to assess and compare the cardioprotective potential of oleuropein (OL), hydroxytyrosol (HT), oleocanthal (OC) and oleanolic Acid (OA), regarding Ischemia/Reperfusion Injury (IRI) and CVD risk factors alleviation. The scope of the study was to design a potent and safe combinatorial therapy for high-cardiovascular-risk patients on a bench-to-bedside approach. We evaluated the IRI-limiting potential of 6-weeks treatment with OL, HT, OC or OA at nutritional doses, in healthy and metabolic syndrome (MS)-burdened mice. Three combinatorial regimens were designed and the mixture with preponderant benefits (OL-HT-OC, Combo 2), including infarct sparing and antiglycemic potency, compared to the isolated compounds, was further investigated for its anti-atherosclerotic effects. In vivo experiments revealed that the combination regimen of Combo 2 presented the most favorable effects in limiting infarct size and hyperglycemia, which was selected to be further investigated in the clinical setting in Chronic Coronary Artery Syndrome (CCAS) patients. Cardiac function, inflammation markers and oxidative stress were assessed at baseline and after 4 weeks of treatment with the OL-HT-OC supplement in the clinical study. We found that OL, OC and OA significantly reduced infarct size in vivo compared to Controls. OL exhibited antihyperglycemic properties and OA attenuated hypercholesterolemia. OL-HT-OA, OL-HT-OC and OL-HT-OC-OA combination regimens were cardioprotective, whereas only OL-HT-OC mitigated hyperglycemia. Combo 2 cardioprotection was attributed to apoptosis suppression, enhanced antioxidant effects and upregulation of antioxidant enzymes. Additionally, it reduced atherosclerotic plaque extent in vivo. OL-HT-OC supplement ameliorated cardiac, vascular and endothelial function in the small-scale clinical study. Conclusively, OL-HT-OC combination therapy exerts potent cardioprotective, antihyperglycemic and anti-atherosclerotic properties in vivo, with remarkable and clinically translatable cardiovascular benefits in high-risk patients.

Return of the Tbx5; lineage-tracing reveals ventricular cardiomyocyte-like precursors in the injured adult mammalian heart.

The single curative measure for heart failure patients is a heart transplantation, which is limited due to a shortage of donors, the need for immunosuppression and economic costs. Therefore, there is an urgent unmet need for identifying cell populations capable of cardiac regeneration that we will be able to trace and monitor. Injury to the adult mammalian cardiac muscle, often leads to a heart attack through the irreversible loss of a large number of cardiomyocytes, due to an idle regenerative capability. Recent reports in zebrafish indicate that Tbx5a is a vital transcription factor for cardiomyocyte regeneration. Preclinical data underscore the cardioprotective role of Tbx5 upon heart failure. Data from our earlier murine developmental studies have identified a prominent unipotent Tbx5-expressing embryonic cardiac precursor cell population able to form cardiomyocytes, in vivo, in vitro and ex vivo. Using a developmental approach to an adult heart injury model and by employing a lineage-tracing mouse model as well as the use of single-cell RNA-seq technology, we identify a Tbx5-expressing ventricular cardiomyocyte-like precursor population, in the injured adult mammalian heart. The transcriptional profile of that precursor cell population is closer to that of neonatal than embryonic cardiomyocyte precursors. Tbx5, a cardinal cardiac development transcription factor, lies in the center of a ventricular adult precursor cell population, which seems to be affected by neurohormonal spatiotemporal cues. The identification of a Tbx5-specific cardiomyocyte precursor-like cell population, which is capable of dedifferentiating and potentially deploying a cardiomyocyte regenerative program, provides a clear target cell population for translationally-relevant heart interventional studies.

Associations between Maternal and Offspring Hair Cortisol Concentrations and Child Behavioral Symptoms in Mother-Child Pairs with Perinatal Mental Disorders.

Maternal perinatal mental disorders (PMD) are associated with developmental and behavioral problems in children, probably mediated by the programming of the hypothalamic-pituitary-adrenal (HPA) axis. Increased cortisol concentrations during the antenatal and perinatal periods have been related to long-term effects on children's behavior and stress response. We aimed to investigate the association of hair cortisol concentrations (HCC) between mothers, with (n = 16) and without PMD (n = 30), and their children, aged between 18 and 48 months. Participants were evaluated with a clinical interview and questionnaires for the Depression Anxiety Stress Scale and the Child Behavior Checklist for ages 1½-5. Maternal and child HCCs were compared between the two groups. Children of the PMD group had increased symptoms of attention deficit hyperactivity disorder. A positive linear association between maternal and child HCC was observed only in the total sample of mother-child dyads and the control group. In the PMD group, children's HCCs were significantly associated with child anxiety/depression symptoms. Aggressive behavior and oppositional/defiant problems correlated significantly with children's own HCCs, and their mother's too. These findings suggest that a chronic dysregulation of maternal and child HPA axis and their associations in the PMD dyads may underlie the linkage among prolonged maternal stress, child behavioral/emotional problems and stress responses.

A Noninvasive Ocular (Tear) Sampling Method for Genetic Ascertainment of Transgenic Mice and Research Ethics Innovation.

Animal models, animal welfare and research ethics are both facilitators and gatekeepers for Big Data generation in genomics and multi-omics R&D. Safeguarding animal welfare is also a research ethics issue that can benefit from technical innovations in biosample collection in particular. Animal welfare draws from the guiding principles of 3R, namely, "Replacement" (methods avoiding the use of animals in research), "Reduction" (methods using fewer animals or derive more information from the same number of animals), and "Refinement" (methods removing or minimizing pain or distress). We report here that noninvasive ocular (tear) sampling for genetic ascertainment of transgenic mice can serve as an innovative ethical safeguard for animal welfare, and as a veritable alternative to the surgical tail biopsies, ear puncture, or blood sampling from the weanling transgenic mice. We compared ocular versus tail biopsy sampling in regard to ascertainment, by genotyping, of apolipoprotein E-deficient (ApoE-/-) transgenic weanling mice (n = 60) by one-round polymerase chain reaction analysis. We found that ocular sampling compares to the results obtained by tail sampling with the obvious benefit of being noninvasive and improving the 3R, especially for the Refinement principle of animal welfare. To place the importance of this new biosample collection approach into further context, transgenic mice research and animal models are at the epicenter of Big Data translation to health innovation. We suggest that ocular sampling is considered and evaluated further in transgenic mice models, not to mention warrant exploration for applications in other types of animal models that require noninvasive biosample collection.

The effect of propranolol on aortic structure and function in normotensive rats.

INTRODUCTION: Beta-blocking agents are widely used for the treatment of many cardiovascular diseases. The effect of these agents, however, on the aortic wall structure and function has not been well defined. The present study was undertaken to investigate the effect of therapy with propranolol on wall structure and aortic function in rats. METHODS: 20 healthy Wistar rats (350-400 g) were assigned to a control group (n=8), with rats receiving only water and food, and an experimental group (n=12), in which 100 mg/kg/day propranolol was administered in the drinking water. Three months after initiation of treatment, aortic pressures and aortic pulse wave velocity (PWV) were measured using high-fidelity Millar catheters. Extensive histopathologic studies were performed in the wall of the descending thoracic aorta. RESULTS: Systolic, mean, diastolic, and pulse pressure were significantly lower in the propranolol-treated rats compared to controls (p<0.05). For any given systolic, mean, and pulse pressure, PWV was greater in the propranolol-treated animals (p<0.05). The heart rate was lower and the response to isoproterenol infusion was less in the propranolol-treated animals. Smooth muscle content was decreased and collagen content was increased in the aortic wall of the propranolol-treated animals compared to controls. CONCLUSIONS: Long-term propranolol administration elicits an increase in PWV adjusted for aortic pressure. This may be related to accumulation of collagen in the aortic wall at the expense of smooth muscle cells. The aortic stiffening may explain some of the reported data, suggesting that the effect of ß-blockade therapy in patients with arterial hypertension may be inferior to other pharmacologic agents.