The Synergistic Anti-Apoptosis Effects of Amniotic Epithelial Stem Cell Conditioned Medium and Ponesimod on the Oligodendrocyte Cells

Multiple sclerosis is a chronic inflammatory and neurodegenerative disease of the central nervous system. The current treatment of Multiple sclerosis is based on anti-inflammatory disease-modifying treatments, which can not regenerate myelin and eventually neurons. So, we need new approaches for axonal protection and remyelination. Amniotic epithelial stem cells amniotic epithelial cells, as a neuroprotective and neurogenic agent, are a proper source in tissue engineering and regenerative medicine. Due to differentiation capability and secretion of growth factors, AECs can be a candidate for the treatment of MS. Moreover, sphingosine-1-phosphate (S1P) receptor modulators were recently approved by FDA for MS. Ponesimod is an S1P receptor-1 modulator that acts selectively as an anti-inflammatory agent and provides a suitable microenvironment for the function of the other neuroprotective agents. In this study, due to the characteristics of AECs, they are considered a treatment option in MS. The conditioned medium of AECs concurrently with ponesimod was used to evaluate the viability of the oligodendrocyte cell line after induction of cell death by cuprizone. Cell viability after treatment by conditioned medium and ponesimod was increased compared to untreated groups. Also, the results showed that combination therapy with CM and ponesimod had a synergistic anti-apoptotic effect on oligodendrocyte cells. The combination treatment with CM and ponesimod reduced the expression of caspase-3, caspase-8, Bax, and Annexin V proteins and increased the relative BCL-2/Bax ratio, indicating inhibition of apoptosis as a possible mechanism of action. Based on these promising results, combination therapy with amniotic stem cells and ponesimode could be a proper alternative for multiple sclerosis treatment

Inhibition of EGF and CoCl2-Induced HIF-1α and VEGF Production in Triple Negative MDA-MB-468 Cells by Umbelliprenin: Unveiling the Role of PI3K/AKT/mTOR and MAPK Pathways

Background and objectives: Triple-negative breast cancer is a significant global health challenge, and there's growing interest in targeting multiple pathways for treatment. Umbelliprenin, derived from herbal sources, has shown anti-tumor potential. This study aimed to assess umbelliprenin's impact on key genes related to proliferation, metastasis, and angiogenesis. Methods: Umbelliprenin, which was synthesized by the Pharmaceutical Research Center (PRC) at Mashhad University of Medical Sciences in Iran, was utilized in this study. The study aimed to investigate the impact of umbelliprenin on EGF and CoCl2-induced signaling in the PI3K/AKT/mTOR and MAPK pathways. Quantitative PCR was employed to assess the expression of EGFR, PI3K, AKT, mTOR, S6K, ERK1, ERK2, 4EBP1, HIF-1α, HIF-1β, VEGF, and VEGFR genes. Additionally, immunoblot assays were conducted to evaluate the levels of VEGF and HIF-1α in MDA-MB-468 cells. Results: The study found that umbelliprenin had cytotoxic effects, with an IC50 value of 152.5 µM. At concentrations of 10 µM and 20 µM, it upregulated genes like EGFR, VEGFR, HIF-1α, VEGF, PI3K, ERK2, and mTOR while downregulating 4EBP1. Umbelliprenin also increased VEGF protein levels. When used on EGF-stimulated cells, it enhanced VEGF and PI3K expression while inhibiting AKT, ERK2, mTOR, and antiproliferative 4EBP1 genes. Notably, VEGF and HIF-1α protein levels remained unchanged. Conversely, umbelliprenin downregulated EGFR, AKT, ERK1/2, HIF-1α, and VEGF in CoCl2-stimulated cells, while elevating 4EBP1 and reducing VEGF and HIF-1α protein levels. Conclusion: Umbelliprenin inhibited MDA-MB-468 cell growth and impacted gene expression related to metastasis and angiogenesis, particularly under conditions of EGFR activation and hypoxia

Protective effect of α-terpineol against impairment of hippocampal synaptic plasticity and spatial memory following transient cerebral ischemia in rats

Objective(s): Cerebral ischemia is often associated with cognitive impairment. Oxidative stress has a crucial role in the memory deficit following ischemia/reperfusion injury. α-Terpineol is a monoterpenoid with anti-inflammatory and antioxidant effects. This study was carried out to investigate the effect of α-terpineol against memory impairment following cerebral ischemia in rats. Materials and Methods: Cerebral ischemia was induced by transient bilateral common carotid artery occlusion in male Wistar rats. The rats were allocated to sham, ischemia, and α-terpineol-treated groups. α-Terpineol was given at doses of 50, 100, and 200 mg/kg, IP once daily for 7 days post ischemia. Morris water maze (MWM) test was used to assess spatial memory and in vivo extracellular recording of long-term potentiation (LTP) in the hippocampal dentate gyrus was carried out to evaluate synaptic plasticity. Malondialdehyde (MDA) was measured to assess the extent of lipid peroxidation in the hippocampus. Results: In MWM test, α-terpineol (100 mg/kg, IP) significantly decreased the escape latency during training trials (

Interference-free Determination of Carbamazepine in Human Serum Using High Performance Liquid Chromatography: A Comprehensive Research with Three-way Calibration Methods

Abstract In the present study, a comprehensive and systematic strategy was described to evaluate the performance of several three-way calibration methods on a bio-analytical problem. Parallel factor analysis (PARAFAC), alternating trilinear decomposition (ATLD), self-weighted alternating trilinear decomposition (SWATLD), alternating penalty trilinear decomposition (APTLD), and unfolded partial least squares combined with the residual bilinearization procedure (U-PLS/RBL) were applied on high performance liquid chromatography with photodiode-array detection (HPLC-DAD) data to quantify carbamazepine (CBZ) in different serum samples. Using the proposed approach, successfully quantification of CBZ in human plasma, even in the presence of diverse uncalibrated serious interfering components was achieved. Moreover, the accuracy and precision of each algorithm for analyzing CBZ in serum samples were compared using root mean square error of prediction (RMSEP), the recovery values and figures of merits and reproducibility of the analysis. Satisfying recovery values for the analyte of interest were obtained by HPLC-DAD on a Bonus-RP column using an isocratic mode of elution with acetonitrile/K2HPO4 (pH = 7.5) buffer solution (45:55) coupled with second-order calibrations. Decreas of the analysis time and less solvent consumption are some of the pluses of this method. The analysis of real samples showed that the modeling of complex chromatographic profiles containing CBZ as the target drug using any of the mentioned algorithms can be potentially benefit drug monitoring in therapeutic research

Minocycline Improves Memory by Enhancing Hippocampal Synaptic Plasticity and Restoring Antioxidant Enzyme Activity in a Rat Model of Cerebral Ischemia-Reperfusion

Introduction: Oxidative stress plays a crucial role in the impairment of synaptic plasticity following cerebral ischemia, ultimately resulting in memory dysfunction. Hence, the applying antioxidant agents could be beneficial in managing memory deficits after brain ischemia. Minocycline is a tetracycline antibiotic with antioxidant effect. The main objective of this work was to assess the minocycline effect on the impairment of synaptic plasticity and memory after cerebral ischemia-reperfusion in rats. Methods: Transient occlusion of common carotid arteries was used to induce ischemia-reperfusion injury in rats. Single or multiple (once daily for 7 days) dose(s) of minocycline were administered before (pretreatment) or after (treatment) brain ischemia. Seven days after ischemia-reperfusion, passive avoidance performance, long-term hippocampal potentiation, and the activity of antioxidant enzymes were assessed.  Results: The passive avoidance test showed that minocycline (20 and 40 mg/kg) significantly increased step-through latency while reducing the duration of staying in a dark chamber in the treatment (but not pretreatment) group. In electrophysiological experiments, the rats treated (but not pretreated) with minocycline (40 mg/kg) showed a significant increase in the amplitude of the field excitatory postsynaptic potentials in the dentate gyrus area of the hippocampus. The treatment (but not pretreatment) with minocycline (20 and 40 mg/kg) resulted in a significant increase in the activity of catalase, glutathione peroxidase, and superoxide dismutase in the hippocampus.  Conclusion: It was determined that minocycline attenuates memory dysfunction after cerebral ischemia-reperfusion in rats by improving hippocampal synaptic plasticity and restoring antioxidant enzyme activity

Role of L-arginine/NO/cGMP/KATP channel signaling pathway in the central and peripheral antinociceptive effect of thymoquinone in rats

Objective(s): Growing evidence demonstrates that L-arginine/NO/cGMP/KATP channel pathway has a modulatory role in pain perception. Previous studies have shown that thymoquinone exerts antinociceptive effects; however, the mechanisms underlying antinociception induced by thymoquinone have not been fully clarified. The aim of the present study was to evaluate the role of L-arginine/NO/cGMP/KATP channel pathway in the central and peripheral antinociceptive effect of thymoquinone in rats.Materials and Methods: Rats were pretreated intraplantarly (IPL) or intracerebroventricularly (ICV) with L-arginine (the NO precursor), l-NAME (an NO synthase inhibitor), SNAP (an NO donor), methylene blue (a guanylyl cyclase inhibitor), glibenclamide (the blocker of KATP channel), and tetraethylammonium (TEA, a Kv channel blocker) before the injection of thymoquinone. Results: Local ipsilateral (20 and 40 μg, IPL) but not contralateral and ICV (4 and 8 μg) administration of thymoquinone caused a dose-dependent and significant antinociception in both early and late phases of the formalin test. Pretreatment of rats with L-arginine (100 μg, IPL or ICV) and SNAP (200 μg, IPL or ICV) increased while l-NAME (100 μg, IPL or 1 μg, ICV) and methylene blue (400 μg, IPL or ICV) decreased the antinociceptive effects of thymoquinone in the formalin test. The administration of TEA (IPL or ICV) did not modify but glibenclamide (50 μg, IPL or ICV) significantly abolished the peripheral and central antinociceptive effects of thymoquinone in both phases of the formalin test. Conclusion: The results of the present study indicate that L-arginine/NO/cGMP/KATP channel pathway participates in the central and peripheral antinociceptive effect of thymoquinone

Nanoliposomal Auraptene: A Comprehensive Study on Preparation, Characterization, Cytotoxicity, and Anti-Angiogenic Potential

Aims: To suppress angiogenesis, auraptene is used in the form of liposome to enhance solubility and effectiveness. Background: Nanoliposomes are spherical nano-sized capsules enclosed by lipid membranes, serving as a biocompatible vehicle to enhance the delivery of therapeutic agents. Objective: The objective of this research is to prepare and characterize nanoliposome-encapsulated auraptene and compare its cytotoxic and anti-angiogenic effects to non-liposomal auraptene. Methods: Liposomal auraptene was formulated using DSPC/DSPG/Cholesterol (molar ratio of 4:1:2) in combination with two different molar ratios of auraptene (0.1 and 0.05). The entrapment efficiency was evaluated using High-Performance Liquid Chromatography (HPLC). Various parameters, including Dynamic Light Scattering (DLS), zeta potential, stability, and release kinetics, were investigated. Subsequently, both liposomal and non-liposomal auraptene, along with bare liposomes, were applied to the MDA-MB-231 cell line for a duration of 72 hours at 37°C at varying concentrations. Cytotoxicity was assessed using the MTT assay. Additionally, the study examined the anti-angiogenic effects on the vessels in the chorioallantoic membrane of chick embryos. Results: The entrapment efficiency of auraptene was found to be satisfactory at 50%. The liposome size ranged from 85 to 241 nm, with a Z-Average of 190.9 nm. The zeta potentials for all formulations were consistently around -55.7, and the Polydispersity Index (PDI) was less than 0.3 for all formulations. The release profile demonstrated approximately 80% drug release over a period of 130 hours. Notably, liposomal auraptene exhibited a significantly lower IC50 value (38.61 [95% Confidence Interval: 30.56 to 48.78]) compared to non-liposomal auraptene (50.36 [95% Confidence Interval: 43.58 to 58.19]) (p = 0.0240). Conclusion: Moreover, the administration of 2.5 and 5 µM of liposomal auraptene led to a reduction in the vessels within the chorioallantoic membrane at the injection site when compared to the control group. In summary, the use of biodegradable nanoliposomal carriers improved the solubility, release profile, and stability of auraptene while demonstrating anticancer and anti-angiogenic properties

Increasing serum albumin in patients with hypoalbuminemia does not inhibit serum angiotensin-converting enzyme activity: Serum Albumin Increase and Serum ACE Activity in Hypoalbuminemic Patients

Introduction: Angiotensin-converting enzyme (ACE) plays a pivotal role in the production of angiotensin II and the inactivation of bradykinin. Recent studies have suggested that human serum albumin may possess ACE-inhibitory properties, serving as a potential endogenous ACE inhibitor that primarily affects serum ACE levels. Interestingly, the infusion of albumin in the postoperative phase of cardiac surgery has been associated with the development of hypotension. Methods: This study aimed to assess serum ACE activity in 27 hypoalbuminemia patients admitted to the ICU before and after a protein-rich diet was administered to raise their serum albumin levels. Serum ACE activity was quantified using raas(HPLC), measuring the formation of hippuric acid, a product generated during the incubation of serum with Hip-His-Leu, a substrate, at 37°C for 30 minutes. Results: In vitro experiments demonstrated that the addition of albumin to human sera led to a significant reduction in ACE activity compared to control groups (P < 0.0001). This reduction was consistent across all serum samples. Specifically, the maximum velocity (Vmax) of ACE activity significantly decreased from 14.90 U/L in the control group to 3.210 U/L in the albumin-added group (P = 0.0262). Notably, there was no significant change in the Michaelis constant (Km) between the control group (0.5263 mM) and the albumin group (0.2742 mM) (P = 0.6763), indicating a non-competitive inhibitory effect of albumin on ACE activity. Interestingly, in this study, elevating serum albumin levels in hypoalbuminemia patients following a protein-rich diet resulted in both ACE inhibition and a slight increase in activity (P = 0.0201). This increase correlated mildly with serum albumin levels across all samples. Conclusions: In conclusion, contrary to in vitro findings, raising serum albumin levels in hypoalbuminemia patients did not further inhibit serum ACE activity