Nitric Oxide Release Research Articles

Vibroacoustic therapy to treat pain in the temporomandibular joint

Pain in the temporomandibular joint (TMJ) affects about 4.8 % of the United States population. Pain is the most common symptom of TMJ and the chief reason patients seek care. Conventional treatments range from non-invasive therapies including physical therapy, dental guards and occlusal splints to interventions such as corticosteroid injections and surgery. At present, the most common challenge to healthcare providers in patients presenting with TMJ is the reduction of pain. Vibroacoustic therapy (VAT) is noninvasive and safe as a potential treatment for TMJ pain. VAT involves low-frequency vibrations from 30 to 120 + Hertz. Biological mechanisms to explain a beneficial effect include enhanced blood circulation, decreased swelling, and promotion of muscle relaxation. VAT also appears to have beneficial effects on skin and bone conduction as well as stimulates relaxation responses and the release of nitric oxide. Clinical studies of VAT to reduce TMJ pain are promising but limited in design and sample sizes. The most reliable design strategy to detect the most plausible small to moderate benefit of VAT on TMJ pain is a randomized trial. This trial should be of sufficient size, dose, and duration. If VAT has a favorable benefit-to-risk ratio, then it would represent a safe, noninvasive, feasible and cost-effective means to treat TMJ.

Achieving Biomedically Desirable Nitric Oxide Release from Glucose Monitor Surfaces Via a Cu-Based Catalyst Coating.

We report the design of a blood-contacting glucose monitor with a nitric oxide (NO)-releasing metal-organic framework (MOF) embedded within the outer polymer layer of a glucose sensor to promote the release of NO from endogenous NO donors. The sensors were tested by using amperometry across a range of glucose concentrations to assess whether the presence of either the MOF or NO decreased the performance of the glucose monitor. Even though signal response was diminished, the sensors maintained a good regression fit (R2 = 0.9944) and a similar dynamic range and reproducibility in the presence of S-nitrosoglutathione.

NIR-Triggered Multifunctional NO Nanoplatform for Conquering Thermal Resistance in Biofilms.

Photothermal treatment (PTT) has emerged as a promising avenue for biofilm elimination, yet its potential drawbacks, such as local hyperpyrexia and bacterial heat resistance, have posed challenges. To address these concerns, an innovative nanoplatform (Au@mSiO2-arg/ICG) is devised that integrates phototherapeutic and gas therapeutic functionalities. This multifaceted nanoplatform is composed of mesoporous silica-coated Au nanorods (Au@mSiO2), supplemented with l-arginine (l-arg) and indocyanine green (ICG), and is engineered for mild temperature PTT aimed at biofilm eradication. Au@mSiO2-arg/ICG nanoparticles (NPs) show excellent antibacterial effects through the generation of nitric oxide (NO) gas, heat, and reactive oxygen species (ROS) under 808nm light irradiation. The ROS generated by ICG initiates a cascade reaction with l-arg, ultimately yielding NO gas molecules. This localized release of NO not only effectively curbs the expression of heat shock proteins 70 mitigating bacterial thermoresistance, but also reduces extracellular polymeric substance allowing better penetration of the therapeutic agents. Furthermore, this nanoplatform achieves an outstanding biofilm elimination rate of over 99% in an abscess model under 808nm light irradiation (0.8 W·cm-2), thereby establishing its potential as a dependable strategy for NO-enhanced mild PTT and antibacterial photodynamic therapy (aPDT) in clinical settings.

Applications of nitric oxide-releasing nanomaterials in dermatology: Skin infections and wound healing

Nitric oxide (NO) is produced in most cells in the skin and is an important regulator of essential cutaneous functions, including responses to UV irradiation, microbial defense, wound healing, melanogenesis and epidermal permeability barrier homeostasis. Harnessing the physiological activities of NO for therapeutic use is difficult because the molecule is highly reactive and unstable. A variety of exogenous NO delivery platforms have been developed and evaluated; however, they have limited clinical applications in dermatology due to instability and poor cutaneous penetration. NO-releasing nanomaterials overcome these limitations, providing targeted tissue delivery, and sustained and controlled NO release. This review provides a comprehensive and up-to-date evaluation of the use of NO-releasing nanomaterials in dermatology for the treatment of skin and soft tissue infections and wound healing.

DEFB114 protein enhances host resistance to fungal infection through the NOD1/2-ATG16L1-NF-κB signaling pathway

The overuse of antibiotics has led to the enhanced resistance of many pathogenic bacteria, posing a threat to human health. Therefore, there is a need to develop green and safe alternatives to antibiotics. Beta-defensins play a crucial role in host defense against pathogens and have multifunctional properties, exerting key roles in innate and adaptive immunity, as well as non-immune processes. In this study, a 210 bp long cDNA sequence of yak DEFB114 gene was amplified and successfully expressed in a prokaryotic system. The DEFB114 protein exhibited significant inhibitory effects on the growth of Aspergillus fumigatus in vitro. When co-cultured with yak macrophages, DEFB114 protein enhanced macrophage phagocytic activity and increased nucleic acid fluorescence intensity (P < 0.05). DEFB114 protein also enhanced the activity of yak macrophages stimulated by inactivated Aspergillus fumigatus spores, increased the release of nitric oxide (NO), and promoted the expression of genes such as γ-actin, Lgals, Man2b, and Capg (P < 0.05). In mice experiments, DEFB114 protein promoted resistance against Aspergillus fumigatus infection, by regulating the NOD1/2-ATG16L1-NF-κB pathway to modulate the host immune response and exert its anti-infective effects. In summary, the yak DEFB114 protein could inhibit the growth of Aspergillus fumigatus and enhance the animal's resistance to pathogenic microorganisms, thereby having significant implications in the treatment and prevention of fungal infections.

Potential Anti-Inflammatory Constituents from Aesculus wilsonii Seeds.

A chemical study of Aesculus wilsonii Rehd. (also called Suo Luo Zi) and the in vitro anti-inflammatory effects of the obtained compounds was conducted. Retrieving results through SciFinder showed that there were four unreported compounds, aeswilosides I-IV (1-4), along with fourteen known isolates (5-18). Their structures were elucidated by extensive spectroscopic methods such as UV, IR, NMR, [α]D, and MS spectra, as well as acid hydrolysis. Among the known ones, compounds 5, 6, 8-10, and 12-16 were obtained from the Aesculus genus for the first time; compounds 7, 11, 17, and 18 were first identified from this plant. The NMR data of 5 and 18 were reported first. The effects of 1-18 on the release of nitric oxide (NO) from lipopolysaccharide (LPS)-induced RAW264.7 cells were determined. The results showed that at concentrations of 10, 25, and 50 μM, the novel compounds, aeswilosides I (1) and IV (4), along with the known ones, 1-(2-methylbutyryl)phloroglucinyl-glucopyranoside (10) and pisuminic acid (15), displayed significant inhibitory effects on NO production in a concentration-dependent manner. It is worth mentioning that compound 10 showed the best NO inhibitory effect with a relative NO production of 88.1%, which was close to that of the positive drug dexamethasone. The Elisa experiment suggested that compounds 1, 4, 10, and 15 suppressed the release of TNF-α and IL-1β as well. In conclusion, this study enriches the spectra of compounds with potential anti-inflammatory effects in A. wilsonii and provides new references for the discovery of anti-inflammatory lead compounds, but further mechanistic research is still needed.

Characterization of an exopolysaccharide synthesized by Lacticaseibacillus rhamnosus B6 and its immunomodulatory activity in vitro

An exopolysaccharide, designated F1, was purified from the fermented milk by Lacticaseibacillus rhamnosus strain B6 (CGMCC No. 13310). F1, with the weight average molecular weight of 1.577 × 106 Da, is consisted of rhamnose, glucose and galactose in a molar ratio of 3.7:1.5: 1. The backbone included 1,3-linked Rha, 1,2,3-linked Rha, 1,2-linked Glc and 1,3-linked Glc residues, with the branching point located at O2 position of 1,2,3-linked Rha residue, and the branch chain composed of terminal linked galactose residue with a pyruvate substituent. F1 could significantly stimulate the phagocytic activity and TNF-α expression in RAW 264.7 macrophages in a dose-dependent manner, and the release of NO at 200 μg/mL as well. F1 at 200 μg/mL could stimulate the expression of the pro-inflammatory cytokine encoding genes including TNF-α and iNOS, but with a negligible upregulating effect on the mRNA expression of IL-10. F1 could up-regulate the expression of NF-κBp65 and skew macrophage polarization towards M1 phenotype. These results suggest F1 elicit an immunomodulatory effect through the NF-κB signaling pathway.

Ultrasound triggered local sequential reactive oxygen species and nitric oxide release for enhanced cerasomal drug delivery

Liposomal drug delivery for tumor therapy was still suffered from low stability, limited delivering efficiency and poor tumor penetration, greatly compromising the therapeutic efficacy. Herein, a stable and ultrasound triggered cerasomal drug delivery system was developed by loading doxorubicin (DOX) into stable liposomal cerasome, which was self-assembled from cerasome-forming lipid (CFL), sonosensitizer of porphyrin lipid (PL), nitric oxide (NO) donor of DSPE-PEG2000-SNO and unsaturated phospholipids DOPC, resulting in DOX@PC-SNO with stable siloxane network and S-nitrosothiol on surface, sonosensitizer in lipid bilayer and DOX in the core. DOX@PC-SNO can prevent drug leakage during blood circulation and efficiently accumulate at the tumor site at 24 h post-injection, while controllably generate reactive oxygen species (ROS) upon local ultrasound irradiation on tumor, which can not only use for SDT, but also operate as a switch for on-demand controlled release of internal DOX and external NO, resulting in specific delivery and high penetration of chemotherapeutic drug into the deep tumor, thus greatly improved antitumor efficacy without observed side effects.

Construction of poly (ionic liquid)-derived gold/silver alloy@nitrogen-doped carbon shell and its application for ratiometric electrochemical detection of nitric oxide

A nitrogen-doped carbon shell loaded with a gold and silver alloy (Au/Ag@NCS) was constructed for highly sensitive electrochemical detection of NO. The Au/Ag@NCS material was prepared by use of SiO2 particles as a template to polymerize imidazolium-based ionic liquids loaded with gold and silver salts, and subsequent carbonization treatment and template removal. The hollow structure of the carbon material acted as a carrier for electrochemical sensing, offering high specific surface area, large pore capacity, robust electron conductivity, and excellent mechanical stability. The inclusion of gold in the composite enhanced its catalytic and sensing capabilities, while silver oxidation was employed as a reference signal for accurate detection. By utilization of the Au/Ag@NCS-modified electrode, a wide detection range from 0.5 nM to 1.05 μM with a low detection limit of 0.32 nM was achieved for NO detection. The electrochemical sensor also exhibited high selectivity and excellent stability. The fabricated sensor was further utilized to explore the release of NO from breast cancer cells, revealing that the electrochemical platform could be regarded as an important method to study the daily tests of NO in clinical application.

Hyperchloremia and Hypernatremia Decreased Microglial and Neuronal Survival during Oxygen-Glucose Deprivation/Reperfusion.

Hyperchloremia and hypernatremia are associated with higher mortality in ischemic stroke, but it remains unclear whether their influence directly contributes to ischemic injury. We investigated the impact of 0.9% sodium chloride (154 mM NaCl), 0.9% sodium acetate (167 mM CH3COONa), and their different combinations (3:1, 2:1, and 1:1) on microglial (HMC-3) and neuronal (differentiated SH-SY5Y) survival during oxygen-glucose deprivation/reperfusion (OGD/R). Further, we assessed the effect of hyperchloremia and hypernatremia-treated and OGD/R-induced HMC-3-conditioned media on differentiated SH-SY5Y cells under OGD/R conditions. We performed cell viability, cell toxicity, and nitric oxide (NO) release assays and studied the alteration in expression of caspase-1 and caspase-3 in different cell lines when exposed to hyperchloremia and hypernatremia. Cell survival was decreased in 0.9% NaCl, 0.9% CH3COONa, combinations of HMC-3 and differentiated SH-SY5Y, and differentiated SH-SY5Y cells challenged with HMC-3-conditioned media under normal and OGD/R conditions. Under OGD/R conditions, differentiated SH-SY5Y cells were less likely to survive exposure to 0.9% NaCl. Expression of caspase-1 and caspase-3 in HMC-3 and differentiated SH-SY5Y cells was altered when exposed to 0.9% NaCl, 0.9% CH3COONa, and their combinations. A total of 0.9% NaCl and 0.9% CH3COONa and their combinations decreased the NO production in HMC-3 cells under normal and OGD/R conditions. Both hypernatremia and hyperchloremia reduced the survival of HMC-3 and differentiated SH-SY5Y cells under OGD/R conditions. Based on the OGD/R in vitro model that mimics human ischemic stroke conditions, it possibly provides a link for the increased death associated with hyperchloremia or hypernatremia in stroke patients.

Berberine attenuates inflammation and oxidative stress and modulates lymphocyte E-NTPDase in acute hyperlipidemia.

Hyperlipidemia is a common clinically encountered health condition worldwide that promotes the development and progression of cardiovascular diseases, including atherosclerosis. Berberine (BBR) is a natural product with acknowledged anti-inflammatory, antioxidant, and metabolic effects. This study evaluated the effect of BBR on lipid alterations, oxidative stress, and inflammatory response in rats with acute hyperlipidemia induced by poloxamer-407 (P-407). Rats were pretreated with BBR (25 and 50 mg/kg) for 14 days and acute hyperlipidemia was induced by a single dose of P-407 (500 mg/kg). BBR ameliorated hypercholesterolemia, hypertriglyceridemia, and plasma lipoproteins in P-407-adminsitered rats. Plasma lipoprotein lipase (LPL) activity was decreased, and hepatic 3-hydroxy-3-methylglutaryl CoA (HMG-CoA) reductase activity was enhanced in hyperlipidemic rats. The expression of low-density lipoprotein receptor (LDL-R) and ATP-binding cassette transporter 1 (ABCA1) was downregulated in hyperlipidemic rats. BBR enhanced LPL activity, upregulated LDL-R, and ABCA1, and suppressed HMG-CoA reductasein P-407-administered rats. Pretreatment with BBR ameliorated lipid peroxidation, nitric oxide (NO), pro-inflammatory mediators (interleukin [IL]-6,IL-1β, tumor necrosis factor [TNF]-α, interferon-γ, IL-4 and IL-18) and enhanced antioxidants. In addition, BBR suppressed lymphocyte ecto-nucleoside triphosphate diphosphohydrolase (E-NTPDase) and ecto-adenosine deaminase (E-ADA) as well as NO and TNF-α release by macrophages isolated from normal and hyperlipidemic rats. In silico investigations revealed the binding affinity of BBR towardLPL, HMG-CoA reductase, LDL-R, PSK9, ABCA1, and E-NTPDase. In conclusion, BBR effectively prevented acute hyperlipidemia and its associated inflammatory responses by modulating LPL, cholesterolgenesis, cytokine release, and lymphocyte E-NTPDase and E-ADA. Therefore, BBR is an effective and safe natural compound that might be employed as an adjuvant against hyperlipidemia and its associated inflammation.

Effect of preparation method of pyrolysis liquid mixed with coal on its ignition and combustion characteristics

Relevance. The need to develop technological solutions to increase the competitiveness of pyrolysis processing of various wastes. Combustion of pyrolysis liquid in a mixture with coal is one of these solutions, which makes it possible to stabilize the properties of the obtained fuel and use in standard energy equipment. Aim. To determine the influence of the method of preparing a mixture of pyrolysis liquid and low-grade coal on its ignition and combustion characteristics, as well as on composition of the released gas-phase products, depending on the temperature of heating medium and concentration of the additive. Methods. Pyrolysis and oxidation characteristics were studied using thermogravimetric analysis, and formal kinetic constants were calculated using the Coates–Radfern method. Samples of mixed fuels were prepared by the methods of homogeneous mixing and surface wetting of pyrolysis liquid of rubber processing and low-grade coal. Ignition and combustion characteristics were determined using an experimental stand, and composition of gas-phase combustion products was determined using once-through gas analyzer. Results. The authors have determined the features of pyrolysis and oxidation of the pyrolysis liquid as well as the values of formal kinetics constants, indicating the physical nature of the factors that defining the rate of these processes. It was found that at 600 °C the ignition of the studied mixtures weakly depended on concentration of the additive, while at 700 and 800 °C the dependence was linear, and the differences between the samples prepared using different methods were insignificant. For samples obtained by the method of surface wetting, combustion of the additive occurred in the gas phase near the surface of the sample. For the samples obtained by the method of uniform mixing, it occurred predominantly in the bulk of the backfill. This led to more intense and complete coal combustion in these compositions due to more uniform releasing of heat and initiation of coal particles. The concentration curves of the NO, CO and CO2 release demonstrated, that behavior of the fuel mixture components was additive in terms of released gas-phase combustion products, as well as the absence of significant nonlinear effects over the entire studied range of temperatures and additive concentrations.

Chickpea Sprouts as a Potential Dietary Support in Different Prostate Disorders-A Preliminary In Vitro Study.

Prostate cancer (PC) and benign prostatic hyperplasia (BPH) are common health problems in the aging male population. Due to the unexplored and unconfirmed impact of food containing isoflavones, like sprouts, on the development of the management of BPH and prostate cancer, we decided to extend the knowledge in this area. We have demonstrated for the first time that chickpea sprouts may play an important role in the chemoprevention of prostate disorders. However, attention should be paid to the isoflavone content in the sprouts, as in our study, chickpea sprouts with a moderate concentration of the compounds, harvested in natural light conditions (CA10L) and blue LED light (CA7B), showed the best scores in terms of their potential towards prostate disorders. Chickpea seeds were grown in LED chambers. The methanol extracts from sprouts were quantitatively defined using the HPLC system. Experiments such as the determination of PSA, 5-α-reductase, and dihydrotestosterone were performed on PNT2 and LNCaP cells. For anti-inflammatory assays (determination of NO, IL-6, and TNF-alpha release), murine RAW264.7 macrophages were used. The role of legume products as a diet element should be deeply evaluated for the development of future dietary recommendations for prostate cancer and BPH prevention.

Accurate Spatio-Temporal Delivery of Nitric Oxide Facilitates the Programmable Repair of Avascular Dense Connective Tissues Injury.

Avascular dense connective tissues (e.g., the annulus fibrosus (AF) rupture, the meniscus tear, and tendons and ligaments injury) repair remains a challenge due to the "biological barrier" that hinders traditional drug permeation and limits self-healing of the injured tissue. Here, accurate delivery of nitric oxide (NO) to penetrate the "AF biological barrier" is achieved thereby enabling programmable AF repair. NO-loaded BioMOFs are synthesized and mixed in a modified polyvinyl alcohol and PCL-composited electrospun fiber membrane with excellent reactive oxygen species-responsive capability (LN@PM). The results show that LN@PM could respond to the high oxidative stress environment at the injured tissue and realize continuous and substantial NO release. Based on low molecular weight and lipophilicity, NO could penetrate through the "biological barrier" for accurate AF drug delivery. Moreover, the dynamic characteristics of the LN@PM reaction can be matched with the pathological microenvironment to initiate programmable tissue repair including sequential remodeling microenvironment, reprogramming the immune environment, and finally promoting tissue regeneration. This tailored programmable treatment strategy that matches the pathological repair process significantly repairs AF, ultimately alleviating intervertebral disc degeneration. This study highlights a promising approach for avascular dense connective tissue treatment through intelligent NO release, effectively overcoming "AF biological barriers" and programmable treatment.

Anti-inflammatory and anti-arthritic potential of Coagulansin-A: in vitro and in vivo studies.

The current work was designed to evaluate the anti-inflammatory and anti-arthritic potential of Coagulansin-A (Coag-A) using mouse macrophages and arthritic mice. In the LPS-induced RAW 264.7 cells, the effects of Coag-A on the release of nitric oxide (NO), reactive oxygen species (ROS), and pro-inflammatory cytokines were analyzed. In addition, the mediators involved in the nuclear factor-kappa B (NF-κB) and nuclear factor erythroid 2-related factor 2 (Nrf2) pathways were evaluated by the RT-qPCR and western blotting. Coag-A did not show significant cytotoxicity in the RAW 264.7 cells in the tested concentration range (1-100 µM). Coag-A significantly inhibited the production of NO, ROS, and key pro-inflammatory cytokines. The anti-inflammatory effects of Coag-A might be through inhibiting the NF-κB pathway and activating the Nrf2 pathway. In the arthritic mouse models, behavioral studies and radiological and histological analyses were performed. We found that the i.p. injection of Coag-A dose-dependently (1-10 mg/kg) reduced the Carrageenan-induced acute inflammation in the mice. In Complete Freund's Reagent-induced arthritic mouse model, Coag-A (10 mg/kg) showed significant anti-inflammatory and anti-arthritic effects in terms of the arthritic index, hematological parameters, and synovium inflammation. After the Coag-A treatment, the bone and tissue damage was ameliorated significantly in the arthritic mice. Moreover, immunohistochemistry of mouse paw tissues revealed a significant reduction in the expression of pro-inflammatory cytokines in the NF-κB pathway, confirming Coag-A's therapeutic potential and mechanism.

Glu and GSH double-unlocked immunomodulatory tri-cascade nanoreactor for biofilm-infected diabetic wounds healing

The treatment of diabetic wounds remains a challenge due to the interaction between hyperglycemia, immune imbalance and bacterial infection. Herein, we developed a Glu and GSH double-unlocked immunomodulatory nanocascade reactor to accelerate diabetic wounds healing which dynamically manages the wound microenvironment by controlling blood glucose levels, restoring immune regulation and eliminating bacteria. On the one hand, ultra-small gold nanoparticles with GOx activity can catalyse glucose at the diabetic wound site, which can be further enhanced by photothermal effects, thus destroying bacterial biofilm structures, reversing wounds pH, and reducing bacterial reproduction. On the other hand, the sustained controlled release of NO at specific sites was achieved by depleting over-expressed GSH in the wounds to restore normal immune regulation, which not only promotes M2 macrophage polarisation and expressions of anti-inflammatory factors such as IL-10 and TGF-β1, but also produces VEGF and CD31 via the PI3K/AKT signalling pathway. Especially, the chemical resonance energy transfer (CRET) strategy was first applied to diabetic wounds treatment, achieving chemical photodynamic therapy (PDT) while reducing the secondary tissue damage caused by catalytically-generated H2O2. In summary, the nanocascade reactor provides a new reference for bacterialy-infected diabetic wounds treatment mode by reducing blood glucose and restoring immune regulation in parallel.

Alkaloids from Fritillaria przewalskii Bulbs and Their Anti-Alzheimer's Disease Activities.

Three undescribed isosteroidal alkaloids, przewalskines A-C (1-3), as well as seven known alkaloids (4-10) were obtained from Fritillaria przewalskii bulbs. Their structures were deduced by extensive HRESIMS, 1D NMR, and 2D NMR analyses, and their bioactivities were evaluated involving the anti-inflammatory and inhibitory potencies on AChE, BChE, and Aβ aggregation. Compound 4 revealed the potent effect on inhibiting Aβ aggregation activity with IC50 value of 33.1 μM, AChE activity with IC50 value of 6.9 μM, and also showed NO release inhibitory acitivity with IC50 value of 32.6 μM. These findings contribute new multi-.target anti-AD agents and embody the chemical diversity of F. przewalskii.

Eugenol's Molecular Warfare against Human Leukemia K562 cells: In Vitro Insights to Chemotherapeutic Potentials

Background: The search for effective anticancer agents has led to an increased focus on natural compounds, given their role as the foundation for many chemotherapeutic drugs. Eugenol (EUG) and Bis-Eugenol (Bis-EUG), derived from clove oil, have shown promise due to their antioxidative, anti-inflammatory, pro-apoptotic, and anti-proliferative properties across various cancer cell lines. This study aims to elucidate the chemotherapeutic potential of EUG and Bis-EUG on human leukemia K562 cells, exploring their ability to modulate apoptosis and cell proliferation. Objective: To assess the in vitro anticancer effects of EUG and Bis-EUG on K562 cell lines through various assays measuring cell viability, apoptosis, gene expression, and nitric oxide release, with a focus on elucidating the mechanisms underlying their anticancer activity. Methods: The study employed an inter-collaborative approach, utilizing MTT assays to determine the IC50 values of EUG and Bis-EUG on K562 cells. Gene expression analysis of pro-apoptotic genes Caspase-3 and Caspase-9 was performed using RT-qPCR. Apoptotic changes were further analyzed through flow cytometry and Hoechst 33258 staining to observe morphological alterations in treated cells. The release of nitric oxide (NO) as an indicator of anti-proliferative activity was measured using a Griess assay. Statistical analysis was conducted using GraphPad Prism Software, Version 9, and Microsoft Excel, with a significance threshold of p ≤ 0.05. Results: The IC50 values for EUG and Bis-EUG were determined to be 16.7 µM and 0.14 mM, respectively. Gene expression profiling revealed a significant increase in Caspase-3 and Caspase-9 expression, indicating the activation of apoptotic pathways. Flow cytometry and Hoechst 33258 staining confirmed the induction of apoptosis, with noticeable changes in cell morphology and nuclear fragmentation. The NO release assay demonstrated a dose-dependent increase in NO levels, highlighting the compounds' anti-proliferative effects. Conclusion: EUG and Bis-EUG exhibit potent anticancer effects on leukemia K562 cells by inducing apoptosis and inhibiting cell proliferation. The significant increase in NO levels and the activation of caspases suggest that these compounds may serve as effective chemotherapeutic agents. Further in vivo studies are warranted to explore their potential in cancer treatment regimens.

Anti-inflammatory activity of seven plant species with potential use as livestock feed additives

Inflammatory conditions and associated oxidative stress are common in livestock. The present study aimed to investigate in depth the anti-inflammatory activity of seven medicinal plants (Dichrostachys cinerea, Ehretia rigida, Salix babylonica, Vachellia erioloba, V. gerrardii, V. sieberiana and V. tortilis) traditionally used to treat inflammatory disorders in South Africa. These plants are being investigated for possible inclusion in animal feed as additives to support health and promote growth. The anti-inflammatory potential of plant extracts was evaluated in terms of inhibition of the 15-lipoxygenase (15-LOX) enzyme and nitric oxide (NO) release in LPS-induced RAW 264.7 macrophages, together with cytotoxicity studies. The effect of plant extracts in modulating the expression of pro-inflammatory mediators including cyclooxygenase (COX), inducible nitric oxide synthase (iNOS), and the cytokine interleukin-6 (IL-6) was evaluated using quantitative reverse transcription polymerase chain reaction (qRT-PCR). In vitro antioxidant activity was investigated using chemical assays. ULPC-MS and ICP-MS were used to detect potentially bioactive phytochemicals and concentrations of essential elements respectively. Methanol and acetone extracts of D. cinerea had the best activity against 15-LOX (IC50 values of 0.80 and 0.64 mg/ml, respectively). The methanol extracts of E. rigida, V. tortilis and V. sieberiana inhibited NO with IC50 of 90.11, 101.52 and 94.11 µg/ml respectively, higher than that of the positive control (IC50 = 30.00 µg/ml). Furthermore, these plants had low cytotoxicity (LC50 ≥ 70 µg/ml) against RAW 264.7 macrophages. Interestingly, E. rigida, V. sieberiana and V. tortilis extracts decreased the expression of iNOS, COX and IL-6. There was a positive correlation between the inhibition of NO release from macrophages by plant extracts and the downregulation of iNOS mRNA levels. There was no significant difference in the antioxidant activity (ABTS) of D. cinerea methanolic extract with IC50 = 5.37 µg/ml compared to the positive controls, Trolox (IC50 = 4.42 µg/ml) and ascorbic acid (IC50 = 4.43 µg/ml). Flavonoids, fatty acids and essential trace elements such as Fe, Zn and Mn were detected in E. rigida, and these are likely to be responsible for the good anti-inflammatory effect of the extracts. This study provides the first evidence of the regulation of pro-inflammatory mediator and cytokine genes by the selected plant extracts. These results support the use of the plants, in particular E. rigida, as potential feed additives to reduce inflammation associated with many diseases.

Endothelial cells signaling and patterning under hypoxia: a mechanistic integrative computational model including the Notch-Dll4 pathway.

Introduction: Several signaling pathways are activated during hypoxia to promote angiogenesis, leading to endothelial cell patterning, interaction, and downstream signaling. Understanding the mechanistic signaling differences between endothelial cells under normoxia and hypoxia and their response to different stimuli can guide therapies to modulate angiogenesis. We present a novel mechanistic model of interacting endothelial cells, including the main pathways involved in angiogenesis. Methods: We calibrate and fit the model parameters based on well-established modeling techniques that include structural and practical parameter identifiability, uncertainty quantification, and global sensitivity. Results: Our results indicate that the main pathways involved in patterning tip and stalk endothelial cells under hypoxia differ, and the time under hypoxia interferes with how different stimuli affect patterning. Additionally, our simulations indicate that Notch signaling might regulate vascular permeability and establish different Nitric Oxide release patterns for tip/stalk cells. Following simulations with various stimuli, our model suggests that factors such as time under hypoxia and oxygen availability must be considered for EC pattern control. Discussion: This project provides insights into the signaling and patterning of endothelial cells under various oxygen levels and stimulation by VEGFA and is our first integrative approach toward achieving EC control as a method for improving angiogenesis. Overall, our model provides a computational framework that can be built on to test angiogenesis-related therapies by modulation of different pathways, such as the Notch pathway.