Background and Purpose: Neuroprotective strategies for stroke remain inadequate. Nanoliposomes comprised of phosphatidylcholine, cholesterol, and monosialogangliosides (nanoliposomes) induced an antioxidant protective response in endothelial cells exposed to amyloid insults. We tested the hypotheses that nanoliposomes will preserve human neuroblastoma (SH-SY5Y) and human brain microvascular endothelial cells viability following oxygen-glucose deprivation (OGD)–reoxygenation and will reduce injury in mice following middle cerebral artery occlusion. Methods: SH-SY5Y and human brain microvascular endothelial cells were exposed to oxygen-glucose deprivation–reoxygenation (3 hours 0.5%–1% oxygen and glucose-free media followed by 20-hour ambient air/regular media) without or with nanoliposomes (300 µg/mL). Viability was measured (calcein-acetoxymethyl fluorescence) and protein expression of antioxidant proteins HO-1 (heme oxygenase-1), NQO1 (NAD[P]H quinone dehydrogenase 1), and SOD1 (superoxide dismutase 1) were measured by Western blot. C57BL/6J mice were treated with saline (n=8) or nanoliposomes (10 mg/mL lipid, 200 µL, n=7) while undergoing 60-minute middle cerebral artery occlusion followed by reperfusion. Day 2 postinjury neurological impairment score and infarction size were compared. Results: SH-SY5Y and human brain microvascular endothelial cells showed reduced viability post–oxygen-glucose deprivation–reoxygenation that was reversed by nanoliposomes. Nanoliposomes increased protein expressions of HO-1, NQO1 in both cell types and SOD1 in human brain microvascular endothelial cells. Nanoliposomes-treated mice showed reduced neurological impairment and brain infarct size (18.8±2% versus 27.3±2.3%, P =0.017) versus controls. Conclusions: Nanoliposomes reduced stroke injury in mice subjected to middle cerebral artery occlusion likely through induction of an antioxidant protective response. Nanoliposome is a candidate novel agent for stroke.
There is a substantial unmet need for the treatment of skeletal muscle mass loss that is associated with aging and obesity-related increases in FFA. Unsaturated FFAs stimulate the inflammatory gene expression in human skeletal myoblasts (SkMs). Farnesol is a hydrophobic acyclic sesquiterpene alcohol with potential anti-inflammatory effects. Here, we created farnesol-loaded small unilamellar (SUVs) or multilamellar lipid-based vesicles (MLVs), and investigated their effects on inflammatory gene expression in primary human skeletal myoblasts. The attachment of SUVs or MLVs to SkMs was tracked using BODIPY, a fluorescent lipid dye. The data showed that farnesol-loaded SUVs reduced FFA-induced IL6 and LIF expression by 77% and 70% in SkMs, respectively. Farnesol-loaded MLVs were less potent in inhibiting FFA-induced IL6 and LIF expression. In all experiments, equal concentrations of free farnesol did not exert significant effects on SkMs. This report suggests that farnesol, if efficiently directed into myoblasts through liposomes, may curb FFA-induced inflammation in human skeletal muscle.
As effective as antibiotics have been since they were first discovered, antibiotic resistance proves to be a worldwide threat, endangering their efficacy. Over the years, there has been a significant decline in development of novel class antibiotics. These two factors have made once easily treated infections difficult to manage. Carbapenem‐resistant Enterobacteriaceae (CRE) are an emerging threat due to their high levels of antibiotic resistance. CRE have become resistant to almost all antibiotics, and cause 23,000 infections per year in the United States. Metals are a vital nutrient in all living organisms including bacteria and, have thus been targeted by antimicrobial agents. Bacteria, like all living organisms, must regulate their metal homeostasis in order to survive and replicate. Therefore, mechanisms able to diminish the availability of the metals to bacterial pathogens can be used to develop new therapeutic drugs. An example of a mechanism of bacterial resistance to antibiotics is the production of inactivating enzymes, such as carbapenemases, an example of which is seen in Klebsiella pneumoniae. Carbapenemases play an important role against b‐lactam antibiotics such as imipenem in Enterobacteriaceae. Carbapenemases are categorized based on their functional and molecular properties. Class B carbapenemases are metallo‐b‐lactamases (MBL) which require zinc in their active site, and are therefore considered metalloenzymes. MBL can hydrolyze all b‐lactams and are not inactivated by b‐lactamase inhibitors. In this study we analyzed antimicrobial susceptibility of K. pneumoniae isolates, KPC 2146, KPC 1705 and KPC 1706 which contain class B, class A and no carbapenemase, respectively. Metal homeostasis of isolates was disrupted by inducing zinc deficiency using TPEN, a metal chelator with high affinity for zinc, and preparing a zinc deficient minimal media. Carbapenemase activity was assessed using CARBA‐NP test and LC‐MS. Bacterial susceptibility of zinc‐deficient isolates to carbapenems was analyzed using spot plating, growth curve, minimum inhibitory concentration and ring of inhibition. Our findings demonstrate zinc dependency of class B carbapenemase in the KPC 2146 isolate, while class A carbapenemase in the KPC 1705 isolate was not affected. Spot plating and growth curve analyses demonstrated that zinc deficiency lowers bacterial growth in KPC 2146 in the presence of imipenem. Zinc deficiency via TPEN addition lowered imipenem, doripenem, ertapenem and meropenem MIC in KPC 2146. Ring of inhibition of KPC 2146 with imipenem was greatly affected by zinc chelation. In conclusion, we demonstrate that disrupting zinc homeostasis affects class B carbapenemase activity, which in turn increases bacterial susceptibility to carbapenems. Support or Funding Information Nika Padyab was supported by funds from the Biomedical Sciences Program at MWU. Drs. Veltri and Hernandez were supported by MWU intramural funds.
Neuroprotective strategies for stroke remain inadequate. Nanoliposomes comprised of phos-phatidylcholine, cholesterol and monosialogangliosides (NL) induced an antioxidant protective response in endothelial cells exposed to amyloid insults. We tested the hypotheses that NL will preserve SH-SY5Y neuroblastoma cell viability following hypoxic injury and will reduce injury in mice following middle cerebral artery occlusion (MCAO). Neuroblastoma were exposed to 20-hour physoxic (5% oxygen) or hypoxic (1% oxygen) condition without or with NL (100 or 300 µg/mL). Viability was measured using calcein-AM fluorescence and SH-SY5Y gene expression of antioxidant proteins heme oxygenase-1 (HO-1), NAD(P)H quinone dehydrogenase 1 (NQO1) and superoxide dismutase 1 (SOD1) were measured by quantitative polymerase chain reaction. C57BL/6J mice were treated with saline (N=8) or NL (10000 ug/mL, N=7) while undergoing 60-minute MCAO followed by reperfusion. Day 2 post-injury neurologic impairment score and infarction size were compared. Neuroblastoma showed reduced viability following hypoxia that was reversed by NL. NL increased gene expression of HO-1, NQO1 and SOD1 versus controls. NL-treated mice showed reduced neurologic impairment and brain infarct size (18.8±2% versus 27.3±2.3%, p=0.017) versus controls. NL reduced stroke injury in mice subjected to MCAO likely through induction of an antioxidant stress response. NL is a candidate novel agent for stroke.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.
hi@scite.ai
10624 S. Eastern Ave., Ste. A-614
Henderson, NV 89052, USA
Copyright © 2024 scite LLC. All rights reserved.
Made with 💙 for researchers
Part of the Research Solutions Family.