Prenatal exposure to crude oil vapor reduces differentiation potential of rat fetal mesenchymal stem cells by regulating ERK1/2 and PI3K signaling pathways: Protective effect of quercetin.

It has been indicated that crude oil vapor (COV) induces tissue damage by several molecular mechanisms. Quercetin (QT) as an important component of food with antioxidant properties has a protective role against cell toxicity caused by many pollutants. However, data related to the adverse effects of crude oil vapor (COV) on stem cell fate and differentiation and the role of quercetin (QT) in protecting stem cells against the toxicity caused by these pollutants is very limited. This study aimed to explore the protective effect of QT against the adverse effects of COV on fetal mesenchymal stem cells (fMSCs) differentiation. Twenty-four pregnant Wistar rats were categorized into 4 groups including the control, COV, COV+QT, and QT. Rats were exposed to COV from gestational day (GD) 0-15 and received QT by gavage. The fMSCs were isolated from fetuses, and cell proliferation, differentiation potential, expression of osteogenesis and adipogenesis-related genes, and phosphorylation of PI3K and ERK1/2 signaling proteins were evaluated. The results showed that COV reduced the proliferation and differentiation of fMSCs through the activation of PI3K and ERK1/2 signaling pathways. Also, COV significantly decreased the expression of osteonectin, ALP, BMP-6, Runx-2, PPARγ, and CREBBP genes in differentiated cells. QT treatment increased the proliferation and differentiation of fMSCs in COV-exposed rats. In conclusion, our findings suggest that prenatal exposure to COV impaired fMSCs differentiation and QT reduced the adverse effects of COV by regulating ERK1/2 and PI3K signaling pathways.

Comparison of incisional, transverse abdominis plane, and rectus sheath blocks in dogs undergoing ovariohysterectomy.

OBJECTIVE: To compare the analgesia provided by incisional (Incisional), transverse abdominis plane (TAP), and rectus sheath (RS) blocks in dogs submitted for ovariohysterectomy (OHE). ANIMALS: 22 female mixed-breed dogs were allocated into 3 treatments of Incisional (n = 7), TAP (n = 7), and RS (n = 8) and underwent OHE from April 4 to December 6, 2022. PROCEDURES: After premedication with acepromazine (0.05 mg/kg) and morphine (0.5 mg/kg), anesthesia was induced (6 mg/kg) and maintained (0.4 mg/kg/min) with propofol. Each dog randomly received either an incisional (blind technique), TAP, or RS (ultrasound-guided) block. Intraoperative analgesia was assessed using cardiorespiratory variables. Postoperative analgesia was evaluated up to 6 hours after the operation with a Short Form of Glasgow Pain Scale (SF-GCPS) and Visual Analog Scale (VAS). Fentanyl was administered as a rescue analgesic when needed. RESULT: During surgery, all data remained within normal limits without any significant differences. Fentanyl was administered to 1 dog in the Incisional and 1 in the TAP. Post-operatively, a single dose of fentanyl was given to 1 dog in the TAP and 1 in the RS. Four dogs in the Incisional and 3 in the RS received both doses of fentanyl. There was no significant difference regarding postoperative rescue analgesia among treatments. CLINICAL RELEVANCE: All 3 techniques demonstrated acceptable intra- and post-operative analgesia efficacy in dogs undergoing OHE. Further studies are warranted to confirm these findings.

Comparison of intravenous regional anaesthesia with lidocaine and ropivacaine in dogs.

The present study was designed to compare the effects of lidocaine and ropivacaine in intravenous regional anaesthesia (IVRA) in dogs. Twelve adult male dogs were used. Under isoflurane anaesthesia, exsanguination was performed in the target forelimb. Then, a blood pressure cuff was encircled around the limb proximal to the elbow joint with a pressure of approximately 150 mmHg above the mean arterial blood pressure. The animals then received one of the two treatments of lidocaine (3 mg/kg) or ropivacaine (1.5 mg/kg) with a final volume of 0.6 mL/kg into the cephalic vein. After 60 min, the anaesthesia was disrupted and the tourniquet was removed using intermittent opening (30 s) and closing (5 min) manner for three times. The results revealed that at 20 and 30 min after the initiation of IVRA, the dogs in ROP showed higher analgesia than LID. A leakage under the tourniquet during IVRA was detected. Tremor and hypersalivation were observed after tourniquet removal in some dogs. It was concluded that ropivacaine might provide a higher quality of anaesthesia than lidocaine in IVRA in dogs. The development of local anaesthetic toxicity is a major concern and should be considered at the time of tourniquet removal.

Dendritic cells inversely regulate airway inflammation in cigarette smoke-exposed mice.

The recruitment and activation of inflammatory cells into the respiratory system is considered a crucial feature in the pathophysiology of chronic obstructive pulmonary disease (COPD). Because dendritic cells (DCs) have a pivotal role in the onset and regulation of immune responses, we investigated the effect of modulating DC subsets on airway inflammation by acute cigarette smoke (CS) exposure. CS-exposed mice (5 days) were treated with fms-like tyrosine kinase 3 ligand (Flt3L) and 120g8 antibody to increase total DC numbers and deplete plasmacytoid DCs (pDCs), respectively. Flt3L treatment decreased the number of inflammatory cells in the bronchoalveolar lavage (BALF) of the smoke-exposed mice and increased these in lung tissue. DC modulation reduced IL-17 and increased IL-10 levels, which may be responsible for the suppression of the BALF cells. Furthermore, depletion of pDCs led to increased infiltration of alveolar macrophages while restricting the presence of CD103(+) DCs. This study suggests that DC subsets may differentially and compartment-dependent influence the inflammation induced by CS. pDC may play a role in preventing the pathogenesis of CS by inhibiting the alveolar macrophage migration to lung and increasing CD103(+) DCs at inflammatory sites to avoid extensive lung tissue damage.

Cigarette smoke differentially modulates dendritic cell maturation and function in time.

BACKGROUND: Dendritic cells (DCs) as professional antigen presenting cells (APCs) play a critical role in the regulation of host immune responses. DCs evolve from immature, antigen-capturing cells, to mature antigen-presenting cells. The relative contribution of DCs to cigarette smoke-induced inflammation is not well documented. In the current study, we investigated a modulatory effect of cigarette smoke extract (CSE) on differentiation, maturation and function of DCs. METHODS: Primary murine DCs were grown from bone marrow cells with GM-CSF. Development of DC was analyzed by expression of CD11c, MHCII, CD86, CD40 and CD83 using flow cytometry. Murine DC's and human L428 cells were co-cultured with CSE for various periods of time. Functional activity was analyzed by measuring FITC-dextran uptake, cytokine production and the ability to stimulate T cell activation in a mixed lymphocyte reaction. RESULTS: Our results show that short-term CSE stimulation (~24 h) influence the maturation status of newly differentiated and immature DCs towards more mature cells as revealed by upregulation of MHCII, CD83, CD86, CD40, reduction in antigen up-take capacity and enhanced secretion of pro-inflammatory (IL-12, IL-6 and TNF-α) cytokines. Interestingly, long-term CSE exposure, time- and concentration-dependently, suppressed the development of functional DCs. This suppression was demonstrated by a decline in CD11c/MHCII, CD83, CD86 and CD40 expression, the production of cytokines and ability to stimulate T lymphocytes. Moreover, CSE significantly suppressed the endocytosis function of mouse DCs which was not due to diminished DC viability. Similar to mouse DCs, long-term co-culturing of the human L428 DC cell line with CSE time-dependently suppressed the expression of CD54. CONCLUSIONS: The present study provides evidence that CSE modulates DC-mediated immune responses via affecting both the function and maturation of DCs. The suppressive effects of cigarette smoke on DC function might lead to impaired immune responses to various infections.

Dendritic cells in pathogenesis of COPD.

COPD (Chronic Obstructive Pulmonary Disease) is an important lung and airway disease which affects the lives of around 200 million people worldwide with an increasing incidence particularly in developing countries. The pathogenesis of COPD is based on the innate and adaptive inflammatory immune response to the inhalation of toxic particles and gases. Although cigarette smoking is the primary cause of this inflammation, many other environmental and occupational exposures contribute to the pathology of COPD. The immune inflammatory changes associated with COPD are linked to a tissue repair and remodeling process that increases mucus production and causes emphysematous destruction of the gas-exchanging surface of the lung. The inflamed airways of COPD patients contain several inflammatory cells including neutrophils, macrophages, T lymphocytes, and dendritic cells (DCs). Little is known about the relative contribution of DCs in the pathogenesis of COPD. However the number of DCs is changed in smokers and COPD patients and cigarette smoke (CS) induces the release of chemokines from DCs that play a role in the pathogenesis of COPD. In this review paper, an overview is presented on the role of DCs and their mediators in the pathogenesis of COPD. The activation of DCs and their signaling in response to CS will also be highlighted and discussed.