Greater Anti-inflammatory Effect Research Articles

Ultrasmall ruthenium nanozyme with RONS scavenging and anti-inflammatory effects for effective prevention of postoperative peritoneal adhesion

Postoperative peritoneal adhesion is one of the most common complications after abdominal and pelvic surgery. Early inflammatory response and oxidative stress induced by peritoneal injury play important roles in adhesion formation. However, current research mainly focuses on developing barrier materials to physically separate injury sites for preventing peritoneal adhesions, but often ignores the pathologic changes promoting adhesion formation, resulting in unsatisfactory outcomes. Herein we report a versatile nanozyme (ultrasmall ruthenium nanoparticles, RuNPs) with great antioxidant and anti-inflammatory effects targeting the intrinsic causes of peritoneal adhesions. RuNPs sufficiently protect mesothelial cells from oxidative stress by scavenging reactive species and suppress inflammatory response via NF-κB pathway blockade. In rats with cecum abrasion-abdominal wall defect, RuNPs alone or combined with barrier material effectively prevent adhesion formation and promote injured tissue healing through alleviating oxidative stress and inflammation. This study not only generates a promising nano-agent for preventing adhesions but also provides a novel therapeutic strategy by targeting the underlying causation of adhesion formation.

Ischemic myocardial inflammatory signaling in starvation versus hypoxia-derived extracellular vesicles: A comparative analysis

BackgroundCoronary artery disease remains a leading cause of death worldwide. Bone mesenchymal stem cell–derived extracellular vesicles (EVs) have shown promise in the setting of myocardial ischemia. Furthermore, the properties of the EVs can be modified via preconditioning of progenitor cells. Previous research from our lab demonstrated a significant decrease in proinflammatory signaling following treatment with EVs derived from starvation preconditioning of human bone mesenchymal stem cells (MVM EVs) in a porcine model of chronic myocardial ischemia. However, rodent models have demonstrated that the use of EVs derived from hypoxia preconditioning of bone mesenchymal stem cells (HYP EVs) may have extended benefits compared to MVM EVs. This study evaluated the effect of HYP EVs on inflammation in a swine model of chronic myocardial ischemia. We hypothesized that HYP EVs would have a greater anti-inflammatory effect than MVM EVs or saline (CON). MethodsYorkshire swine fed a standard diet underwent placement of an ameroid constrictor to the left circumflex artery. Two weeks later, the animals received intramyocardial injection of saline (CON; n = 6), starvation-derived EVs (MVM; n = 10), or hypoxia-derived EVs (HYP; n = 7). After 5 weeks, myocardial perfusion was assessed, and left ventricular myocardial tissue was harvested. Protein expression was measured using immunoblotting. Data were analyzed via the Kruskal-Wallis test or one-way analysis of variance based on the results of a Shapiro-Wilk test. Coronary perfusion was plotted against relative cytokine concentration and analyzed with the Spearman rank-sum test. ResultsHYP EV treatment was associated with decreased expression of proinflammatory markers interleukin (IL)-6 (P = .03), Pro-IL-1ß (P = .01), IL-17 (P < .01), and NOD-like receptor protein 3 (NLRP3; P < .01) compared to CON. Ischemic tissue from the MVM group showed significantly decreased expression of pro-inflammatory markers NLRP3 (P < .01), IL-17 (P < .01), and HLA class II histocompatibility antigen (P < .01) compared to CON. The MVM group also had decreased expression of anti-inflammatory IL-10 (P = .01) compared to CON counterparts. There were no significant differences in expression of tumor necrosis factor-α, interferon-γ, IL-12, Toll-like receptor-2, and nuclear factor kappa-light-chain-enhancer of activated B cells in either group . There was no correlation between coronary perfusion and cytokine concentration in the MVM or HYP groups, either at rest or with pacing. ConclusionsHYP EVs and MVM EVs appear to result in relative decreases in the degree of inflammation in chronically ischemic swine myocardium, independent of coronary perfusion. It is possible that this observed decrease may partially explain the myocardial benefits seen with both HYP and MVM EV treatment.

Long-term exercise training and inflammatory biomarkers in healthy subjects: a meta-analysis of randomized controlled trials.

This meta-analysis investigated the effect of long-term exercise training (ET) including aerobic, resistance, and multicomponent ET on the levels of inflammatory biomarkers in randomized controlled trials (RCTs) involving healthy subjects. We searched seven databases for articles until May 1st, 2023. A random-effect meta-analysis, subgroup analysis, meta-regressions as well as trim and fill method were conducted using STATA 16.0. Thirty-eight studies were included in the meta-analysis, involving 2,557 healthy subjects (mean age varies from 21 to 86 years). Long-term ET induced significantly decreased in the levels of interleukin-6 (IL-6) (SMD -0.16, 95% CI -0.30 to -0.03, p = 0.017), C-reactive protein (CRP) (SMD -0.18, 95% CI -0.31 to -0.06, p = 0.005), as well as tumor necrosis factor alpha (TNFα) (SMD -0.43, 95% CI -0.62 to -0.24, p < 0.001). Subgroup analysis revealed that Long-term ET conducted for more than 12 weeks and exercise of moderate intensity had greater anti-inflammatory effects. Meta-regression analysis showed that the reduction in CRP level induced by long-term ET was weakened by increasing exercise intensity. Long-term ET induced significant anti-inflammatory effects in healthy subjects. Long-term ET-induced anti-inflammatory effects were associated with exercise of moderate intensity and training conducted for more than 12 weeks.Systematic Review Registration: https://www.crd.york.ac.uk/PROSPERO/# myprospero, PROSPERO, identifier CRD42022346693.

81-LB: Novel Microsphere Technology Reducing Injection Site Reaction in the Development of Long-Acting GLP-1 RA Formulation

Microspheres are typically used for long-acting injectable formulation, but often the injection site reactions are reported as side effects. This can reduce patient compliance with medication despite the benefit of reducing the number of administrations. Side effects at the injection site are mostly derived from inflammatory reactions. Therefore, it is necessary to minimize inflammatory reactions for the successful development of long-acting diabetes treatments, including GLP-1 receptor agonists. The purpose of this study is to evaluate the injection site reaction in the placebo, semaglutide and tirzepatide microspheres manufactured using the conventional microsphere technology and to investigate the injection site reaction improvement in the placebo, semaglutide and tirzepatide microspheres manufactured using novel technology by adding anti-inflammatory agent, dexamethasone. Skin samples were collected from the injection site of SD rats on days 3 and 8 after the administration of microspheres with or without dexamethasone. The collected skin was stained with H&amp;E to determine infiltration of inflammatory cells, angiogenesis, and fibrosis and was graded. When microspheres manufactured using the conventional method were subcutaneously administered to rats, infiltration of inflammatory cells and fibrous tissue formation occurred on both days 3 and 8. Higher dexamethasone content in the microsphere showed a tendency for the lesser occurrence of inflammation and greater anti-inflammatory effect in the test group. Surprisingly, improvement in the injection site reaction for microspheres containing dexamethasone was observed even at systemic dexamethasone concentrations much lower than the therapeutic level. In conclusion, this novel microsphere containing a small amount of dexamethasone shows potential for the development of a long-acting GLP-1 microsphere injectable for diabetes treatment with reduced injection site reaction. Disclosure Y. Kim: None. E. Seol: None. Funding Ministry of Health &amp; Welfare, Republic of Korea (HI21C1732)

The Mechanism of Cannabichromene and Cannabidiol Alone Versus in Combination in the Alleviation of Arthritis-Related Inflammation.

Patients suffering from arthritis have limited treatment options for nonoperative management. In search of pain relief, patients have been taking over-the-counter cannabinoids. Cannabidiol (CBD) and cannabichromene (CBC) are minor cannabinoids with reported analgesic and anti-inflammatory properties and have been implicated as potential therapeutics for arthritis-related pain. To this end, we utilized a murine model to investigate the effectiveness of and mechanism by which CBC alone, CBD alone, or CBD and CBC in combination may provide a reduction in arthritis-associated inflammation. Forty-eight mice were included in the study, which were separated into 4 groups: control group (n = 12), treatment with CBD alone (n = 12), treatment with CBC alone (n = 12), and treatment with CBD + CBC (n = 12). We induced inflammation in each mouse utilizing the collagen-induced arthritis model. At scheduled timepoints, mice were clinically assessed for weight gain, swelling, and arthritis severity. In addition, inflammation-associated serum cytokine levels were analyzed for each animal. Thirty-five of 48 mice survived the duration of the study resulting in the following group numbers: control group (n = 8), treatment with CBD alone (n = 9), treatment with CBC alone (n = 9), and treatment with CBD + CBC (n = 9). Animals treated with CBC and CBD + CBC showed significant weight gain between 3 and 5 weeks. Irrespective of treatment, regression analysis comparing all cytokine measurement and physical outcomes found a significant positive correlation between levels of 5 individual cytokines and both arthritis scores and swelling. Animals treated with CBD + CBC showed a significant decrease in swelling between 3 and 5 weeks compared with the control group. Cannabinoid treatment selectively affected the gene expression of eotaxin and lipopolysaccharide-induced CXC chemokine with combined treatment of CBC + CBD. Treatment with cannabinoids resulted in decreased clinical markers of inflammation. Further, the anti-inflammatory effect of CBC and CBD in conjunction was associated with a greater anti-inflammatory effect than either minor cannabinoid alone. Future work will elucidate the possibility of synergistic or entourage effects of minor cannabinoids used in combination for the treatment of arthritis-related pain and inflammation.

Effective anti-inflammatory phenolic compounds from dandelion: identification and mechanistic insights using UHPLC-ESI-MS/MS, fluorescence quenching and anisotropy, molecular docking and dynamics simulation

This novel study identifies the effective anti-inflammatory phenolic compounds in dandelion and provides mechanistic insights into their interactions with receptor proteins (toll‐like receptor 4, TLR4; co‐receptor myeloid differentiation protein-2, MD-2) using UHPLC-ESI-MS/MS, lipopolysaccharide (LPS)-stimulated THP-1 cell line, fluorescence quenching and anisotropy, molecular docking (single ligand and multi-ligand docking) and molecular dynamics simulation. A 50 % aqueous methanol extract had a greater anti-inflammatory effect and higher chicoric acid content, compared with the 100 % water and 100 % methanol extracts. Chicoric acid, chlorogenic acid, methylophiopogonone A, caffeic acid, gallic acid monohydrate and 4'-O-demethylbroussonin A had relatively high binding energies and contents in all extracts. Chicoric acid competed with chlorogenic acid, 4'-O-demethylbroussonin A and quercetin for MD-2. Among dandelion’s phenolics, chicoric acid most effectively hindered TLR4-MD-2 complex formation, with a quenching constant of 0.62 × 106 L/mol for MD-2 or TLR4 at 320 K, and binding energies of –6.87 and –5.97 kcal/mol, respectively, for MD-2 and TLR4.

Synbiotics and Surgery: Can Prebiotics and Probiotics Affect Inflammatory Surgical Outcomes?

Prebiotics, probiotics, and synbiotics have received increasing attention over the years for their beneficial impact on the gut microbiome and for their systemic anti-inflammatory effects. They have also been shown to improve surgical outcomes. Here, we review the inflammatory effects of surgery as well as the data which suggests a benefit of prebiotics, probiotics, and synbiotics taken in the perioperative period. Synbiotics and fermented foods may have an even greater anti-inflammatory effect than probiotics or prebiotics alone. Recent data suggest that the anti-inflammatory effects and microbiome changes brought on by prebiotics, probiotics, and synbiotics have the potential to improve surgical outcomes. We highlight the potential to alter systemic inflammation, surgical and hospital-acquired infections, colorectal cancer formation, recurrence, and anastomotic leak. Synbiotics could also impact metabolic syndrome. Prebiotics, probiotics, and especially synbiotics may be extremely beneficial when taken in the perioperative period. Even short-term gut microbiome pre-habilitation could alter surgical outcomes significantly.

Amination Potentially Augments the Ameliorative Effect of Curcumin on Inhibition of the IL-6/Stat3/c-Myc Pathway and Gut Microbial Modulation in Colitis-Associated Tumorigenesis.

Epigallocatechin gallate and tetrahydrocurcumin are aminated as colonic metabolites, preserving their bioactivities and improving their capabilities. We compared the bioactivities of unaminated (CUR) and aminated (AC) curcumin in inflammatory colitis-associated tumorigenesis. The anti-inflammatory and anticancer capabilities of CUR and AC were evaluated using RAW264.7 and HT29 cell lines, respectively. An azoxymethane/dextran sodium sulfate-induced colitis-associated carcinogenesis mouse model was used with CUR and two-dose AC interventions. AC had a greater anti-inflammatory effect but a similar anticancer effect as CUR in vitro. CUR and low-dose AC (LAC) significantly preserved colon length and reduced tumor number in vivo. Both CUR and LAC inhibited activation of the protein kinase B (AKT)/nuclear factor kappa B (NF-κB) signaling pathway, its downstream cytokines, and the interleukin (IL)-6/signal transducer and activator of transcription 3 (STAT3)/c-myelocytomatosis oncogene (c-MYC) pathway. However, only LAC significantly preserved E-cadherin, reduced N-cadherin, and facilitated beneficial gut microbial growth, including Akkermansia and Bacteroides, potentially explaining AC's better ameliorative effect at low than high doses.

Metformin therapy as a strategy to compensate anti-VEGF resistance in patients with diabetic macular edema.

Diabetic macular edema (DME) is the complication of diabetic retinopathy, the leading cause of vision loss among diabetic patients. Metformin is the main antidiabetic treatment. It is preferable for its great anti-angiogenic and anti-inflammatory effects. Anti-vascular endothelial growth factor (VEGF) therapy is the preferable treatment for DME despite its lack of convincing results in some patients. To assess whether the combination of metformin and anti-VEGF drugs may decrease the risk of anti-VEGF resistance among DME patients. We included DME patients with a central retinal thickness (CRT) ≥ 250 μm who consecutively underwent at least 3 anti-VEGF therapies from January 1, 2020, to December 30, 2021. Anti-VEGF resistance was defined as persistent macular edema with decreased CRT ≤ 25% after 3 anti-VEGF injections. 109 patients were considered for this research, of whom 65 (59.6%) were resistant to anti-VEGF therapy. The mean CRT of the non-metformin group decreased from 344.88 ± 129.48 to 318.29 ± 123.23 (20.85%) and from 415.64 ± 144.26 to 277.11 ± 99.25 (31.51%) (P = .031) in the metformin group. Moreover, the metformin group had fewer resistant patients than the non-metformin, 24 (45.3%) versus 41 (73.2%). Furthermore, a considerable gain in visual acuity was observed in both groups, with a BCVA gain of 40.41% in the metformin group and 39.9% in the non-metformin group. Metformin may be combined with an anti-VEGF drug to minimize the risk of anti-VEGF resistance among DME patients. Moreover, it can serve to design effective therapeutic deliveries.

Synthesis and Biological Evaluation of Cassane Diterpene (5α)-Vuacapane-8(14), 9(11)-Diene and of Some Related Compounds.

A set of thirteen cassane-type diterpenes was synthesized and an expedient synthetic route was used to evaluate 14-desmethyl analogs of the most active tested cassane. The anti-inflammatory activities of these 13 compounds were evaluated on a lipopolysaccharide (LPS)-activated RAW 264.7 cell line by inhibition of nitric oxide (NO) production, some of them reaching 100% NO inhibition after 72 h of treatment. The greatest anti-inflammatory effect was observed for compounds 16 and 20 with an IC50 NO of 2.98 ± 0.04 μg/mL and 5.71 ± 0.14 μg/mL, respectively. Flow-cytometry analysis was used to determine the cell cycle distribution and showed that the inhibition in NO release was accompanied by a reversion of the differentiation processes. Moreover, the anti-cancer potential of these 13 compounds were evaluated in three tumor cell lines (B16-F10, HT29, and Hep G2). The strongest cytotoxic effect was achieved by salicylaldehyde 20, and pterolobirin G (6), with IC50 values around 3 μg/mL in HT29 cells, with total apoptosis rates 80% at IC80 concentrations, producing a significant cell-cycle arrest in the G0/G1 phase, and a possible activation of the extrinsic apoptotic pathway. Additionally, initial SAR data analysis showed that the methyl group at the C-14 positions of cassane diterpenoids is not always important for their cytotoxic and anti-inflammatory activities.

Is Dutasteride a Therapeutic Alternative for Amyotrophic Lateral Sclerosis?

Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease that is characterized by the loss of upper and lower motor neurons (MNs) in the cerebral cortex, brainstem and spinal cord, with consequent weakness, atrophy and the progressive paralysis of all muscles. There is currently no medical cure, and riluzole and edaravone are the only two known approved drugs for treating this condition. However, they have limited efficacy, and hence there is a need to find new molecules. Dutasteride, a dual inhibitor of type 1 and type 2 5α-reductase (5AR) enzymes, the therapeutic purposes of which, to date, are the treatment of benign prostatic hyperplasia and androgenic alopecia, shows great anti-ALS properties by the molecular-topology methodology. Based on this evidence, this review aims to assess the effects of dutasteride on testosterone (T), progesterone (PROG) and 17β-estradiol (17BE) as a therapeutic alternative for the clinical improvement of ALS, based on the hormonal, metabolic and molecular pathways related to the pathogenesis of the disease. According to the evidence found, dutasteride shows great neuroprotective, antioxidant and anti-inflammatory effects. It also appears effective against glutamate toxicity, and it is capable of restoring altered dopamine activity (DA). These effects are achieved both directly and through steroid hormones. Therefore, dutasteride seems to be a promising molecule for the treatment of ALS, although clinical studies are required for confirmation.

Aktivitas Antiinflamasi Ekstrak Kayu Bajakah (Spatholobus littoralis Hask)

Bajakah wood (Spatholobus littoralis Hassk) is a native plant of Kalimantan which is widely used by the community to treat diarrhea, dysentery, aches, and pains, wound medicine and is even believed to be a cancer drug. Research on this plant is still very limited. The content of flavonoids is thought to act as an anti-inflammatory. The purpose of this study was to determine the effectiveness of Kayu Bajakah (Spatholobus littoralis Hassk) as an anti-inflammatory in male rats induced by carrageenin. The method used in this study was an experimental method using 21 male white rats aged 2-3 months weighing 150-250 grams which were divided into 7 groups, namely the normal group, the CMC Na group, the diclofenac sodium group 1.35 mg/day. 200 g BW and 4 doses of Kayu Bajakah ethanol extract: 50, 100, 200, and 400 mg/kg BW orally. Fifteen minutes after treatment, mice were injected with 1% carrageenin subplantar. The volume of edema was measured with a plestimometer every 60 minutes for 4 hours. Anti-inflammatory was measured by comparing the percentage value of edema inhibition in the control and treatment groups. The results showed that the four treatment groups had a significantly different total volume of edema at 240' minutes with a significance value less than 0.05. The greatest anti-inflammatory effect was at a dose of 400 mg/kg BW at the 240th minute with a percentage of 87.65%. Kayu Bajakah (Spatholobus littoralis Hassk) ethanol extract has anti-inflammatory activity.

The protective effects of S14G-humanin (HNG) against mono-sodium urate (MSU) crystals- induced gouty arthritis

ABSTRACT Gout is a common and complex form of arthritis that has brought great inconveniences to the normal lives of patients. It is reported that oxidative stress and nod-like receptor family protein 3 (NLRP3) inflammasome-mediated inflammatory reactions are involved in the pathogenesis of gout arthritis. S14G-humanin (S14G-HNG) is a modified peptide of HNG with higher inhibitory activity on the accumulation and deposition of Aβ. Recently, S14G-HNG has been reported to exert great anti-inflammatory effects. The present study proposed to explore the possible therapeutic property of S14G-HNG against gout arthritis. An animal model was established by stimulation with mono-sodium urate (MSU) crystals, followed by treatment with colchicine and S14G-HNG, respectively. The elevated Gait score promoted synovitis score and activated myeloperoxidase (MPO) observed in MSU crystals-treated mice were significantly reversed by colchicine and S14G-HNG. Bone marrow-derived macrophages (BMDMs) were isolated from mice and stimulated with MSU crystals, followed by being treated with 25 and 50 μM S14G-HNG. The increased mitochondrial reactive oxygen species (ROS) and Malondialdehyde (MDA) levels, upregulated NADPH oxidase-4 (NOX-4), activated NLRP3 inflammasome, and elevated production of inflammatory factors in MSU crystals-treated BMDMs were dramatically reversed by S14G-HNG, accompanied by the upregulation of sirtuin type-1 (SIRT1). Lastly, the protective effects of S14G-HNG against MSU crystals-induced NLRP3 inflammasome activation were significantly abolished by the knockdown of SIRT1. In conclusion, our data reveal that S14G-HNG could possess potential benefits against MSU crystals-induced gout arthritis, with colchicine displaying a better effect.

The protein corona modulates the inflammation inhibition by cationic nanoparticles via cell-free DNA scavenging

A central paradigm in nanomedicine is that when synthetic nanoparticles (NPs) enter the body, they are immediately cloaked by a corona of macromolecules (mostly proteins) that mediates the role of the physico-chemical properties in the NP biological functions (the “coronation paradigm”). In this work, we focused on the assessment of the “coronation paradigm” for cationic NPs (cNPs) used as rheumatoid arthritis (RA) drugs due to their ability to scavenge cell-free DNA (cfDNA). We fabricated series of cNPs uniformly coated with single or di-hydroxyl groups and different types of amino groups and showed that hydroxylated nanoparticles displayed a prolonged retention in inflamed joints and greater anti-inflammatory effect in collagen-induced arthritis (CIA) rats than the non-hydroxylated analogues. Especially, the cNPs with secondary amines and a di-hydroxyl shell showed the best performance among the tested cNPs. Proteomic analysis showed that the cNPs with a di-hydroxyl shell adsorbed less opsonin proteins than the cNPs carrying mono hydroxyl groups and non-hydroxylated ones, which may provide a mechanistic explanation for the different biodistribution profiles of cNPs. Thus, this study suggests that the protein corona mediates the effects of the surface chemistry on the fate and functions of cNPs as anti-RA drugs.

Effectiveness of Turmeric and Extra Virgin Olive Oil in the Management of IL 6 and IL 10 in Healthy Mice

Background: Anti-Inflammatory properties of turmeric and extra virgin olive oil have numerous health benefits. They exert and promote anti-inflammatory actions, moreover, each gives benefits that go beyond reducing inflammation.&#x0D; Objective: The aim of our study is to evaluate and compare the immunomodulatory effect of each turmeric and extra virgin olive oil (EVOO) alone or in combination.&#x0D; Methods: We demonstrated the serum level of pro-inflammatory cytokines IL-6 and IL-10 in healthy male Swiss albino mice (C57BL/6 strain) after oral intake of curcumin and extra virgin olive oil for six weeks by ELISA assay. &#x0D; Results: Our results showed that dietary intake of turmeric and EVOO had a reducing effect on serum IL-6 level and could increase the production of IL-10 level significantly. Inflammatory responses of turmeric and EVOO combination appear to be more effective on health by inhibition of IL 6 and induce IL 10 production. &#x0D; Conclusion: Anti-inflammatory effect of turmeric and EVOO can play an important role in modulating the level of IL 6 and IL-10. Moreover, co-administration of them having a greater anti-inflammatory effect.

Diclofenac N-Derivatives as Therapeutic Agents with Anti-Inflammatory and Anti-Cancer Effect.

A series of diclofenac N-derivatives (2, 4, 6, 8c, 9c, 10a-c) were synthesized in order to test their anti-cancer and anti-inflammatory effects. The anticarcinogen activity has been assayed against three cancer cell lines: HT29, human colon cancer cells; Hep-G2, human hepatic cells; and B16-F10, murine melanoma cells. First, we determined the cytotoxicity of the different compounds, finding that the most effective compound was compound 8c against all cell lines and both compounds 4 and 6 in human Hep-G2 and HT29 cell lines. Compounds 4 and 8c were selected for the percentage of apoptosis determination, cell cycle distribution, and mitochondrial membrane potential measure because these products presented the lowest IC50 values in two of the three cancer cell lines assayed (B16-F10 and HepG2), and were two of the three products with lowest IC50 in HT29 cell line. Moreover, the percentages of apoptosis induction were determined for compounds 4 and 8c, showing that the highest values were between 30 to 60%. Next, the effects of these two compounds were observed on the cellular cycle, resulting in an increase in the cell population in G2/M cell cycle phase after treatment with product 8c, whereas compound 4 increased the cells in phase G0/G1, by possible differentiation process induction. Finally, to determine the possible apoptosis mechanism triggered by these compounds, mitochondrial potential was evaluated, indicating the possible activation of extrinsic apoptotic mechanism. On the other hand, we studied the anti-inflammatory effects of these diclofenac (DCF) derivatives on lipopolysaccharide (LPS) activated RAW 264.7 macrophages-monocytes murine cells by inhibition of nitric oxide (NO) production. As a first step, we determined the cytotoxicity of the synthesized compounds, as well as DCF, against these cells. Then, sub-cytotoxic concentrations were used to determine NO release at different incubation times. The greatest anti-inflammatory effect was observed for products 2, 4, 8c, 10a, 10b, and 9c at 20 µg·mL−1 concentration after 48 h of treatment, with inhibition of produced NO between 60 to 75%, and a concentration that reduces to the 50% the production of NO (IC50 NO) between 2.5 to 25 times lower than that of DCF. In this work, we synthesized and determined for the first time the anti-cancer and anti-inflammatory potential of eight diclofenac N-derivatives. In agreement with the recent evidences suggesting that inflammation may contribute to all states of tumorigenesis, the development of these new derivatives capable of inducing apoptosis and anti-inflammatory effects at very low concentrations represent new effective therapeutic strategies against these diseases.

Intrapleural Steroid Instillation for Critically Ill Patients With Covid-19 Severe Adult Respiratory Distress Syndrome.

Within a very short period, outbreaks of a novel coronavirus disease (COVID-19) have affected 183 countries worldwide. COVID-19 resolves acutely, but it has resulted in thousands of deaths, with an overall fatality rate of 2% to 5% (1). Among the fatal cases, over 50% of patients died from respiratory failure with massive alveolar damage due to virus-activated “cytokine storm syndrome” (2). Of these patients, 67% developed adult respiratory distress syndrome (ARDS), 29% developed acute kidney injury, 23% developed cardiac injury, and 29% experienced liver dysfunction (3). The intensive care unit (ICU) mortality rate among those who required invasive mechanical ventilation was 86% (4), and the mortality of patients who received extracorporeal membrane oxygenation was also extremely high (5). Until now, there have been no proven effective specific treatment strategies and so methylprednisolone may be the best option for COVID-19 patients who develop ARDS (6). Although interferon alfa-2b, antiviral therapy, antibiotics for the prevention of secondary infection, and methylprednisolone were all administered to attenuate lung inflammation, histological examination still revealed bilateral diffuse alveolar damage with hyaline membrane formation in an autopsy of a patient (7). According to the imaging changes and pathological findings associated with ARDS, a tubular and enlarged appearance of the pulmonary vessels with a sudden caliber reduction seen within ground glass opacity lesions showing partial consolidation and fibrosis should be expected in severe cases of COVID-19 (8, 9). Fortunately, corticosteroids have a greater anti-inflammatory effect when administered intrapleurally, rather than via the intravenous route, because the permeability of the damaged visceral pleural membrane increases during pulmonary edema and steroid treatment, with the characteristic of fat-solubility, further permits infiltration into the lung parenchyma. Glucocorticoids can easily pass through the visceral pleura and bind to steroid hormone receptors, thereby allowing them to reach the injured areas and avoiding the problems of a ventilation–perfusion mismatch and the need for a shunt via intratracheal instillation or intravenous instillation. Glucocorticoids can normalize the ion pump (Na-K pump) and increase the synthesis of Na+/K+-ATPase, thus contributing to reductions in lung edema and optimization of oxygenation because alveolar fluid clearance is impaired in the majority of patients with ARDS (10). In COVID-19 severe refractory ARDS, more aggressive management is required to reduce the inflammatory reaction and consequent lung injury. The results of our study provide more evidence supporting the efficacy of intrapleural steroid instillation (IPSI) in the treatment of ARDS with multiple organ failure (11). This alternative treatment resulted in significant improvement in oxygenation and survival benefits after 5 days of treatment. The dosing schedule of intrapleural methylprednisolone was initially 40 mg q6 h in both pleural cavities, but the dose was reduced on the basis of chest radiography appearance, inspired oxygen concentration demand, and positive end-expiratory pressure of the ventilator. Thoracic catheterization was easily performed because COVID-19 patients usually had significant pleural effusion. Although managing COVID-19 pneumonia is currently a major issue for ICU healthcare providers around the world, IPSI will greatly improve the ability to respond effectively to current needs. Our recommendation in this severe case of COVID-19 might help physicians implement suitable therapeutic strategies for similar severe patients and reduce mortality.

REDUCTION OF OXIDATIVE STRESS OF PULMONARY MEMBRANES DURING LYMPHOTROPIC ADMINISTRATION OF ISONIAZID

The article presents the results of a clinical study of the severity of oxidative stress in the treatment with isoniazid on the material of bronchoalveolar flushes. The object of the study was patients with infiltrative pulmonary tuberculosis who had their bronchoalveolar lavage (BAS) removed during FBS. 19 patients received lymphotropic administration of isoniazid (group 1), 21 patients received intramuscular injection of isoniazid by needle-jet method (group 2), 20 patients were in the 3rd control group with traditional intramuscular administration of isoniazid as part of standard 4 ABP regimens. Lymphotropic injections were performed sequentially in the area of the jugular fossa, behind the xiphoid process, and in the parasternal zone. Group 2 patients were injected with a 10% isoniazid solution in an amount of 1.0 ml into the intercostal space in the projection of the inflammatory focus using a semi-automatic combined ISI-1 acupuncture injector. The content of total protein, malondialdehyde (the final product of lipid peroxidation) and isonicotinic acid hydrazide (non-metabolized GINK, which is part of the anti-tuberculosis drug isoniaziada) was studied in BAS using biochemical methods. Total protein and MDA in patients with tuberculosis are at a high level, reflect the intensity of inflammation and decrease with lymphotropic and intra-pulmonary administration of isoniazid. The content of GINK in BAS is higher with its lymphotropic administration. Based on the study of these tests and the content of GINK in bronchoalveolar flushes, it can be argued that local and regional lymphotropic administration of isoniazid has a greater anti-inflammatory effect on the membranes of the lungs and tracheobronchial tree than intramuscular administration.

Serratiopeptidase: A drug without clear benefits.

Serratiopeptidase is a proteolytic enzyme produced by serratia species E15, isolated from the intestine of silkworm. It has become one of the most widely used medications by both physicians and surgeons and is said to have efficacy in reducing inflammation via the breakdown of various inflammatory mediators. [1] Serratiopeptidase is mostly prescribed in inflammatory illnesses and in some non-inflammatory disorders albeit less commonly due to its analgesic effects. It is even used very commonly in the post-operative period to improve healing and to reduce inflammation of the surgical wound. [2]&#x0D; However, an online literature search on Medline, Cochrane library and Google scholar using the search words serratiopeptidase, serrapeptase, serralysin, serratiapeptase and proteolytic enzymes retrieved only small-scale, inadequately structured, studies reflecting some anti-inflammatory effects of serratiopeptidase, but most of these failed to demonstrate significant benefits from the drug.[3] Serratiopeptidase has a lower efficacy as compared to the NSAIDs as an analgesic but demonstrated greater anti-inflammatory effects than the NSAIDs.[4] Therefore, it is still not sufficiently efficacious to be used solely and rather, is almost always used in combination with NSAIDs to reduce pain symptoms. [5]&#x0D; As a matter of fact, one of the authors’ of this letter had significant experience with the drug in treating seronegative spondyloarthropathy. The drug was used for about a week but no improvement was noted in the reduction of symptoms and the person remained severely disabled. Switching to meloxicam significantly showed good response.&#x0D; In the light of above reports and observations, there is a dire need of conducting well-designed clinical trial-based studies for a wider spectrum of conditions and limit its use until its benefits and/or contraindications are substantially proven [3] Continuous...

Eldecalcitol Inhibits LPS-Induced NLRP3 Inflammasome-Dependent Pyroptosis in Human Gingival Fibroblasts by Activating the Nrf2/HO-1 Signaling Pathway.

PurposePeriodontitis is a major chronic oral disease that is accelerated by activation of the NLRP3 inflammasome and the resulting pyroptosis. According to recent studies, active vitamin D and its analogs have been reported to have great anti-inflammatory effects. However, the anti-inflammatory mechanism of a newly found vitamin D analog, eldecalcitol (ED-71), is still unclear. This study investigates whether ED-71 could protect human gingival fibroblasts (HGFs) from LPS-induced pyroptosis and, if so, determine its underlying mechanism.MethodsAfter HGFs were treated with LPS alone or with LPS and ED-71, their viability was measured by CCK8 assay. The degrees of inflammation and pyroptosis were measured via LDH assay, H2O2 assay, fluorescent staining, flow cytometry, and Western blots. Intracellular ROS, Hoechst 33,342, and PI stains were assessed with a fluorescence microscope. ROS inhibitor NAC, NLRP3 inhibitor MCC950, and Nrf2 inhibitor ML385 were added to further clarify the mechanism.ResultsLPS induced cytotoxicity in HGFs, as shown by CCK8 assay. LPS also increased intracellular ROS, H2O2 levels, release of LDH, and expression of the pyroptosis-related proteins NLRP3, caspase-1, and IL-1β. NAC and MCC950 reduced LPS-induced NLRP3, caspase-1, and IL-1β. Pretreatment with ED-71 effectively inhibited the LPS-induced pyroptosis and was associated with activation of the Nrf2/HO-1 signaling pathway. This beneficial effect of ED-71 was suppressed by ML385.ConclusionThis study demonstrates the therapeutic effect of ED-71 on LPS-induced NLRP3 inflammasome-dependent pyroptosis in HGFs and further reveals that ED-71 can inhibit pyroptosis by activating the Nrf2/HO-1 pathway. Our results thus suggest that ED-71 is a potential candidate for the treatment of periodontitis.