CD200 as a Potential New Player in Inflammation during Rotator Cuff Tendon Injury/Repair: An In Vitro Model.

Rotator cuff tendon (RCT) disease results from multifactorial mechanisms, in which inflammation plays a key role. Pro-inflammatory cytokines and tendon stem cell/progenitor cells (TSPCs) have been shown to participate in the inflammatory response. However, the underlying molecular mechanism is still not clear. In this study, flow cytometry analyses of different subpopulations of RCT-derived TSPCs demonstrate that after three days of administration, TNFα alone or in combination with IFNγ significantly decreases the percentage of CD146+CD49d+ and CD146+CD49f+ but not CD146+CD109+ TSPCs populations. In parallel, the same pro-inflammatory cytokines upregulate the expression of CD200 in the CD146+ TSPCs population. Additionally, the TNFα/IFNγ combination modulates the protein expression of STAT1, STAT3, and MMP9, but not fibromodulin. At the gene level, IRF1, CAAT (CAAT/EBPbeta), and DOK2 but not NF-κb, TGRF2 (TGFBR2), and RAS-GAP are modulated. In conclusion, although our study has several important limitations, the results highlight a new potential role of CD200 in regulating inflammation during tendon injuries. In addition, the genes analyzed here might be new potential players in the inflammatory response of TSPCs.

Pre-Treatment of Swine Oviductal Epithelial Cells with Progesterone Increases the Sperm Fertilizing Ability in an IVF Model.

Mammalian spermatozoa are infertile immediately after ejaculation and need to undergo a functional modification, called capacitation, in order to acquire their fertilizing ability. Since oviductal epithelial cells (SOECs) and progesterone (P4) are two major modulators of capacitation, here we investigated their impact on sperm functionality by using an IVF swine model. To that, we treated SOECs with P4 at 10, 100, and 1000 ng/mL before the coincubation with spermatozoa, thus finding that P4 at 100 ng/mL does not interfere with the cytoskeleton dynamics nor the cells' doubling time, but it promotes the sperm capacitation by increasing the number of spermatozoa per polyspermic oocyte (p < 0.05). Moreover, we found that SOECs pre-treatment with P4 100 ng/mL is able to promote an increase in the sperm fertilizing ability, without needing the hormone addition at the time of fertilization. Our results are probably due to the downregulation in the expression of OVGP1, SPP1 and DMBT1 genes, confirming an increase in the dynamism of our system compared to the classic IVF protocols. The results obtained are intended to contribute to the development of more physiological and efficient IVF systems.

Hormones residues in bovine animals: Sampling, analysis and health risk assessment.

The use of hormones for breeding animal livestock has been banned since 1981 under the Council Directive 81/602/EC. So far, each country should monitor the use of anabolic hormones in animal production to protect the consumer's health against these unwanted residues. This paper presents the research results on steroid and non-steroid hormones residues carried out in Lebanon from 2018 to 2020. Using a newly developed and validated LC-MS/MS method, the detection and the quantification of hormones in bovine matrices were done. The targeted matrices were muscle, liver, kidney, and bile. A total of two-hundred and forty-seven samples were collected from different slaughterhouses located in six different cities in Lebanon. Interestingly, only four hormones were found: testosterone, progesterone, epitestosterone, and 6 propyl 2thiouracil. Based on the obtained data, the estimated daily intake, hazard quotient, and hazard index were calculated to evaluate an exposure assessment.

Human Immune System Diseasome Networks and Female Oviductal Microenvironment: New Horizons to be Discovered.

Human hypofertility and infertility are two worldwide conditions experiencing nowadays an alarming increase due to a complex ensemble of events. The immune system has been suggested as one of the responsible for some of the etiopathogenic mechanisms involved in these conditions. To shed some light into the strong correlation between the reproductive and immune system, as can be inferred by the several and valuable manuscripts published to date, here we built a network using a useful bioinformatic tool (DisGeNET), in which the key genes involved in the sperm-oviduct interaction were linked. This constitutes an important event related with Human fertility since this interaction, and specially the spermatozoa, represents a not-self entity immunotolerated by the female. As a result, we discovered that some proteins involved in the sperm-oviduct interaction are implicated in several immune system diseases while, at the same time, some immune system diseases could interfere by using different pathways with the reproduction process. The data presented here could be of great importance to understand the involvement of the immune system in fertility reduction in Humans, setting the basis for potential immune therapeutic tools in the near future.

Two-Player Game in a Complex Landscape: 26S Proteasome, PKA, and Intracellular Calcium Concentration Modulate Mammalian Sperm Capacitation by Creating an Integrated Dialogue-A Computational Analysis.

Recent experimental findings suggest the involvement of the 26S proteasome, the main protease active in eukaryotic cells, in the process that leads mammalian sperm to become fully fertile, so-called capacitation. Unfortunately, its role in male gametes signaling is still far from being completely understood. For this reason, here, we realized a computational model, based on network theory, with the aim of rebuilding and exploring its signaling cascade. As a result, we found that the 26S proteasome is part of a signal transduction system that recognizes the bicarbonate ion as an input terminal and two intermediate layers of information processing. The first is under the control of the 26S proteasome and protein kinase A (PKA), which are strongly interconnected, while the latter depends on intracellular calcium concentrations. Both are active in modulating sperm function by influencing the protein phosphorylation pattern and then controlling several key events in sperm capacitation, such as membrane and cytoskeleton remodeling. Then, we found different clusters of molecules possibly involved in this pathway and connecting it to the immune system. In conclusion, this work adds a piece to the puzzle of protease and kinase crosstalk involved in the physiology of sperm cells.

LC-MS/MS method for the determination of hormones: Validation, application and health risk assessment in various bovine matrices.

A new method, using liquid chromatography coupled to mass spectrometry (LC-MS/MS) for the detection of fourteen natural and synthetic hormones in muscles, was validated in other bovine matrices (liver, kidney, bile and hair) according to the Decision Commission 2002/657/EC. As result, this method demonstrates good linearity (R2 > 0.99) as well as accuracy with coefficients of variation for repeatability and reproducibility lower than 23%. Moreover, the values of decision limit (CCα) and detection capability (CCß) were determined indicating values ranging from 0.13 to 0.86 µg/kg and 0.25-1.72 µg/k for the majority of analytes. Recovery rate in the different matrices varied from 51.5 to 107%. Indeed, this method has been successfully applied to detect anabolic hormones in eighty-eight samples (muscle, liver, kidney, and bile) collected from different local slaughterhouses. Results showed that progesterone was found in 30 samples at concentrations ranging from 0.11 to 11.7 µg/kg, while testosterone was detected in 34 samples at concentrations ranging from 0.5 to 9.52 µg/kg. All bile samples contain epi-testosterone at concentration ranging from 0.89 to 280 µg/kg. These obtained data were used to calculate the estimated daily intake, hazard quotient and hazard index as exposure assessment.