High Levels Of Nitric Oxide Research Articles

Effect of mitoTEMPO on Redox Reactions in Different Body Compartments upon Endotoxemia in Rats.

Mitochondrial ROS (mitoROS) control many reactions in cells. Biological effects of mitoROS in vivo can be investigated by modulation via mitochondria-targeted antioxidants (mtAOX, mitoTEMPO). The aim of this study was to determine how mitoROS influence redox reactions in different body compartments in a rat model of endotoxemia. We induced inflammatory response by lipopolysaccharide (LPS) injection and analyzed effects of mitoTEMPO in blood, abdominal cavity, bronchoalveolar space, and liver tissue. MitoTEMPO decreased the liver damage marker aspartate aminotransferase; however, it neither influenced the release of cytokines (e.g., tumor necrosis factor, IL-4) nor decreased ROS generation by immune cells in the compartments examined. In contrast, ex vivo mitoTEMPO treatment substantially reduced ROS generation. Examination of liver tissue revealed several redox paramagnetic centers sensitive to in vivo LPS and mitoTEMPO treatment and high levels of nitric oxide (NO) in response to LPS. NO levels in blood were lower than in liver, and were decreased by in vivo mitoTEMPO treatment. Our data suggest that (i) inflammatory mediators are not likely to directly contribute to ROS-mediated liver damage and (ii) mitoTEMPO is more likely to affect the redox status of liver cells reflected in a redox change of paramagnetic molecules. Further studies are necessary to understand these mechanisms.

Distinguished role of MerTk and Axl receptors-mediated efferocytosis in alveolar macrophages on pulmonary mucosal regulation

Abstract Efferocytosis is paramount to regulating homeostasis and inflammation. The TAM receptor family (Tyro3, Axl and MerTk) mediates efferocytosis and simultaneously inhibits pro-inflammatory pathways. Here we investigated how MerTk and Axl regulate pulmonary mucosa homeostasis and inflammation, using silicosis as a study model. During homeostasis, lung cells and alveolar macrophages (AMs) from wild-type (WT) mice showed high expression of Axl, MerTk, and Gas6. Although we have found increased numbers of total airway cells, lower levels of TGF-b and IL-10, and higher levels of nitric oxide in the BALFs from both Axl −/−and MerTk −/−compared to WT mice, enhanced numbers of lung total cells, higher numbers of AMs, monocytes, and neutrophils were found in BALFs and lungs of MerTk −/−compared to Axl −/−or WT mice. We also found increased expression of CXCL1, CXCL2, TNF-a, and IL-6, and enhanced numbers of MHCII +AMs in the lungs of MerTk −/−compared to Axl −/−or WT mice. Furthermore, AMs from Axl −/−mice upregulated the expression of MerTk receptor. During silicosis, we found decreased pulmonary function, increased levels of TGF-b and CXCL1, as well as increased number of AMs and neutrophils in silicotic (SIL)-Axl mice compared to the SIL-MerTk or SIL-WT groups. In addition, SIL-Axl showed a reduced number of CD206 +AMs and an enhanced number of apoptotic, late apoptotic, and necrotic AMs compared to SIL-MerTk or SIL-WT groups. Finally, we found that both SIL-WT and SIL-MerTk upregulated mRNA levels for Axl compared to its PBS groups. Collectively, our data suggest that MerTk and Axl receptor-mediated efferocytosis are dedicated to regulating homeostasis and controlling inflammation in the lungs, respectively. CNPq, CAPES and FAPERJ

Abstract 3314: Circulatory altruistic stem cells as a prognostic biomarker for platinum based chemotherapy in oral cancer

Abstract Background: Head & neck cancer is the 4th common cancer, and most of them are diagnosed late, andtreated with cisplatin monotherapy. In these subsets of patients, immunotherapy has been tried, butwithout major success [1]. We speculate that circulating altruistic mesenchymal stem cells may serve asbiomarker for immunosuppression. Previously, we showed that oral cancer CSCs reprogram MSCs toaltruistic phenotype [2, 3], an embryonic stem cell like phenotype having niche defending ability [5].Here, we propose to test the immunosuppressive ability of ASCs, and their detection in the circulation ofhead & neck cancer subjects undergoing platinum based chemotherapy. Method: The immunosuppression test: immunosuppressive secretory factors, mixed lymphocytereaction (MLR) assay, and a Boyden chamber based co-culture assay with NK cells [4]. Next, weobtained circulating CD271+/CD45- cells of stage IV head and neck cancer subjects (n=20) (5). Kaplan-Meier survival analysis and log-rank test to find association between the presence of circulatory ASCsand overall survival (OS). Result: CSCs including primary tumor obtained CSCs reprogram CD271+ MSCs to ASC phenotype;these cells secrete high level of Nitric oxide, IDO, TGF-beta, IL-10 and PGE-2. ASCs exhibited markedimmunosuppression in the MLR test. In Boyden chamber assay, ASCs markedly decreased the numberof NKG2D+ NK cells, and CD8+ T cells by 4-5 fold, whereas increased the CD4+/FoxP3/CD25- T-reg cells by 3-fold . Importantly, these 12 out of 20 patients showed poortreatment response to platinum therapy. The OS at 6 months was 24.6% for circulating ASC-positivepatients and 47.4% for circulating ASC- negative patients (log-rank test,). Conclusion: The circulatory ASCs could be a promising biomarkers for immunotherapy.

Abstract 3447: NOS inhibition reverses epithelial-to-mesenchymal transition and synergizes with alpelisib in metaplastic breast cancer

Abstract Metaplastic breast cancer (MpBC) is a rare and highly chemoresistant breast cancer subtype, with a median survival of 8 months after metastatic disease, and no standard therapeutic options. MpBCs are enriched for epithelial-to-mesenchymal transition (EMT)/cancer stem cell (CSC) markers, produce high levels of nitric oxide (NO), and have a hyperactive phosphoinositide 3-kinase (PI3K) signaling pathway. NO activates multiple oncogenic pathways, such as PI3K and transforming growth factor beta (TGFβ), a regulator of EMT. Therefore, we hypothesized that pan-NOS inhibitor NG-monomethyl-l-arginine (L-NMMA) could augment the efficacy of α-specific PI3K inhibitor alpelisib in MpBC in vitro and in vivo models. Immunostaining analysis revealed that MpBC PDX tumors had elevated co-expression of iNOS and pAkt (60% vs 23%, p=0.04) relative to triple-negative breast cancer (TNBC) PDX tumors. MpBC PDX tumors had higher RPL39 A14V (66% vs 4.7%, p< 0.00) and PIK3CA hotspot mutation rates (50% vs 19.1%, p=0.31) than TNBC PDX tumors. L-NMMA was synergistic with alpelisib in MpBC cell lines with PIK3CA/PIK3R1 mutations and antagonistic in PIK3CA-wild type and PTEN-deleted models. In vivo evaluation using MpBC PDX tumors found that L-NMMA augmented the efficacy of alpelisib in reducing tumor volume in PIK3CA-mutated MpBC PDX models. Transcriptomic analysis found gene sets associated with EMT reversal, such as the formation of cornified envelope (NES = 2.04 Nom p<0.00) and keratinization pathway (NES = 2.06, Nom p<0.00), were enriched pathways in MpBC PDX tumors that responded to combination therapy. Pharmacological/genomic inhibition of iNOS reversed EMT in MpBC cells, by decreased expression of Zeb1, TGFβ, Snail, Vimentin, and increased expression of E-cadherin and ZO-1 in immunoblotting analysis. MpBC cells with NOS2 knockout acquired an epithelial-like cellular morphology, and this reversal of EMT rendered MpBC cells more sensitive to alpelisib and taxane-chemotherapy. MpBC PDX tumors that responded to combination therapy also exhibited a reversal in EMT, with a decrease in aldehyde dehydrogenase (ALDH1), a CSC marker. Combination therapy also reduced tumor-initiating ability, enhanced chemosensitivity, and improved overall survival in MpBC PDX models. These studies paralleled a phase 1b/2 clinical trial with L-NMMA+taxane chemotherapy in a cohort of anthracycline-refractory MpBC patients (NCT02834403). The clinical benefit rate was 40% (6/15), overall response rate was 20% (3/15), and one patient achieved a pathologic complete response. Relative to baseline tumors, the responder end-of-treatment tumors had undergone reversal of EMT, with decreased expression of iNOS and ALDH1. We find that combining L-NMMA and alpelisib is a novel therapeutic strategy to treat MpBC, and combination therapy is being tested in a first multicenter phase 2 study for patients with MpBC. Citation Format: Tejaswini P. Reddy, Akshjot Puri, Liliana Guzman-Rojas, Christoforos Thomas, Wei Qian, Jianying Zhou, Hong Zhao, Xiaoxian Li, Bijan Mahboubi, Adrian Oo, Cho Young-Jae, Baek Kim, Jose Thaiparambil, Maria Florencia Chervo, Roberto Rosato, Camila Ayerbe, Noah Giese, Stacy Moulder, Helen Piwnica-Worms, Funda Meric-Bernstam, Jenny C. Chang. NOS inhibition reverses epithelial-to-mesenchymal transition and synergizes with alpelisib in metaplastic breast cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3447.

Lutein Encapsulated in PLGA–Phospholipid Nano-Carrier Effectively Mitigates Cytokines by Inhibiting Tumor Necrosis Factor TNF-α and Nuclear Factor NF-κB in Mice Retina

Lutein, a photo- and thermo-labile macular pigment, prevents the retina from suffering ocular inflammation with its antioxidant and anti-inflammatory activity. However, its biological activity is poor due to poor solubility and bioavailability. Therefore, we developed a PLGA NCs (+PL), (poly (lactic-co-glycolic acid) nanocarrier with phospholipid) to improve the biological availability and bioefficacy of lutein in the retina of lipopolysaccharide (LPS)-induced lutein-devoid (LD) mice. The effect of lutein-loaded NCs with/without PL was studied in comparison with micellar lutein. The induction of inflammation by LPS significantly increased the production of nitrites in the LPS-induced group, revealing higher levels of nitric oxide (NO) in the serum (760%) and retina (891%) compared to the control group. Malondialdehyde (MDA) levels in the serum (93%) and retina (205%) of the LPS-induced group were higher compared to the control group. LPS induction resulted in increased protein carbonyls in the serum (481%) and retina (487%) of the LPS group compared to the control group. Further, to conclude, lutein-PLGA NCs (+PL) effectively down-regulated inflammatory complications in the retina.

The Arabidopsis Phytoglobin 2 mediates phytochrome B (phyB) light signaling responses during somatic embryogenesis.

During the light induction of somatic embryogenesis, phyB-Pfr suppresses Phytoglobin 2, known to elevate nitric oxide (NO). NO depresses Phytochrome Interacting Factor 4 (PIF4) relieving its inhibition on embryogenesis through auxin. An obligatory step of many in vitro embryogenic systems is the somatic-embryogenic transition culminating with the formation of the embryogenic tissue. In Arabidopsis, this transition requires light and is facilitated by high levels of nitric oxide (NO) generated by either suppression of the NO scavenger Phytoglobin 2 (Pgb2), or its removal from the nucleus. Using a previously characterized induction system regulating the cellular localization of Pgb2, we demonstrated the interplay between phytochrome B (phyB) and Pgb2 during the formation of embryogenic tissue. The deactivation of phyB in the dark coincides with the induction of Pgb2 known to reduce the level of NO; consequently, embryogenesis is inhibited. Under light conditions, the active form of phyB depresses the levels of Pgb2 transcripts, thus expecting an increase in cellular NO. Induction of Pgb2 increases Phytochrome Interacting Factor 4 (PIF4) suggesting that high levels of NO repress PIF4. The PIF4 inhibition is sufficient to induce several auxin biosynthetic (CYP79B2, AMI1, and YUCCA 1, 2, and 6) and response (ARF5, 8, and 16) genes, conducive to the formation of the embryonic tissue and production of somatic embryos. Auxin responses mediated by ARF10 and 17 appear to be regulated by Pgb2, possibly through NO, in a PIF4-independent fashion. Overall, this work provides a new and preliminary model integrating Pgb2 (and NO) with phyB in the light regulation of in vitro embryogenesis.

Antimicrobial peptide cathelicidin LL-37 preserves intestinal barrier and organ function in rats with heat stroke

Global warming increases the incidence of heat stroke (HS) and HS causes the reduction of visceral blood flow during hyperthermia, leading to intestinal barrier disruption, microbial translocation, systemic inflammation and multiple organ failure. Cathelicidin LL-37 exhibits antimicrobial activities, helps innate immunity within the gut to maintain intestinal homeostasis, and augments intestinal wound healing and barrier function. Thus, we evaluated the effects and possible mechanisms of cathelicidin LL-37 on HS. Wistar rats were placed in a heating-chamber of 42 ̊C to induce HS. Changes in rectal temperature, hemodynamic parameters, and survival rate were measured during the experimental period. Blood samples and ilea were collected to analyze the effects of LL-37 on systemic inflammation, multiple organ dysfunction, and intestinal injury. Furthermore, LS174T and HT-29 cells were used to assess the underlying mechanisms. Our data showed cathelicidin LL-37 ameliorated the damage of intestinal cells induced by HS. Intestinal injury, systemic inflammation, and nitrosative stress (high nitric oxide level) caused by continuous hyperthermia were attenuated in HS rats treated with cathelicidin LL-37, and hence, improved multiple organ dysfunction, coagulopathy, and survival rate. These beneficial effects of cathelicidin LL-37 were attributed to the protection of intestinal goblet cells (by increasing transepithelial resistance, mucin-2 and Nrf2 expression) and the improvement of intestinal barrier function (less cyclooxygenase-2 expression and FITC-dextran translocation). Interestingly, high cathelicidin expression in the ileal samples of inflammatory bowel disease patients was associated with better clinical outcome. These results suggest that cathelicidin LL-37 could prevent heat stress-induced intestinal damage and heat-related illnesses.

Low level of antioxidant capacity biomarkers but not target overexpression predicts vulnerability to ROS-inducing drugs

Despite a strong rationale for why cancer cells are susceptible to redox-targeting drugs, such drugs often face tumor resistance or dose-limiting toxicity in preclinical and clinical studies. An important reason is the lack of specific biomarkers to better select susceptible cancer entities and stratify patients. Using a large panel of lung cancer cell lines, we identified a set of “antioxidant-capacity” biomarkers (ACB), which were tightly repressed, partly by STAT3 and STAT5A/B in sensitive cells, rendering them susceptible to multiple redox-targeting and ferroptosis-inducing drugs. Contrary to expectation, constitutively low ACB expression was not associated with an increased steady state level of reactive oxygen species (ROS) but a high level of nitric oxide, which is required to sustain high replication rates. Using ACBs, we identified cancer entities with a high percentage of patients with favorable ACB expression pattern, making it likely that more responders to ROS-inducing drugs could be stratified for clinical trials.

The Association of High Sensitivity C-reactive protein and Nitric Oxide with Elevated Blood Pressure during Dialysis in Intradialytic Hypertension

BACKGROUND: Hemodialysis (HD) is a procedure to replace part of kidney function in patients with end-stage kidney disease. Hemodynamic disturbances during HD may include elevated blood pressure (BP). It is reported that about 5–15% of patients undergoing regular HD have increased BP during HD. This condition is called intradialytic hypertension (IDH). Increased high sensitivity C-reactive protein (hs-CRP) levels are associated with decreased endothelial dilatation response of blood vessels. Nitric Oxide (NO) is one of the mediators that act as a vasodilator to regulate vascular pressure. Endothelial dysfunction is one of the factors thought to contribute to the incidence of IDH. AIM: This study aimed to determine the association of hs-CRP and NO with elevated BP in IDH. METHODS: This observational cross-sectional study was conducted at Prof. Dr. R.D. Kandou General Hospital, Manado, from November 2021 to April 2022. Participants were selected through consecutive sampling methods. The levels of hs-CRP and NO were examined in all patients. The correlation between hs-CRP and NO was performed using the Spearman test, while the association of hs-CRP and NO with elevated BP was performed using the Fisher- exact test or Chi-square test. RESULTS: Forty patients were included in this study. The median hs-CRP, NO, and systolic BP (SBP) of the patients were 6.55 mg/dL (0.50–43.5), 27.77 mg/dL (3.65–72.19), and 20 mmHg (10–30), respectively. The correlation analysis showed that there was a strong significant negative correlation between hs-CRP levels and NO (r = −0.680, p = 0.000). Based on the Chi-Square or Fisher Exact test, there was a significant association between hs-CRP levels and elevated SBP (OR = 5.06; 95% CI = 1.095–23.44; p = 0.040), as well as between NO levels and elevated SBP (OR = 17.14; 95% CI = 3.063–95.938; p = 0.000). CONCLUSION: There was a significant association between hs-CRP and NO levels with elevated SBP in end-stage renal disease (ESRD) patients with IDH. ESRD on hemodialysis (ESRD on R-HD) patients with hs-CRP ≤3 mg/dL or NO levels <25 mg/dL tends to have increased SBP ≥20 mmHg after HD compared to ESRD patients with low hs-CRP or high NO levels.

Association of Single-Nucleotide Polymorphisms Rs2779249 (chr17:26128581 C>A) and Rs rs2297518 (chr17: chr17:27769571 G>A) of the NOS2 Gene with Tension-Type Headache and Arterial Hypertension Overlap Syndrome in Eastern Siberia.

Inducible nitric oxide (NO) synthase (iNOS), encoded by the NOS2 gene, promotes the generation of high levels of NO to combat harmful environmental influences in a wide range of cells. iNOS can cause adverse effects, such as falling blood pressure, if overexpressed. Thus, according to some data, this enzyme is an important precursor of arterial hypertension (AH) and tension-type headache (TTH), which are the most common multifactorial diseases in adults. The purpose of this study was to investigate the association of rs2779249 (chr17:26128581 C>A) and rs2297518 (chr17: chr17:27769571 G>A) of the NOS2 gene with TTH and AH overlap syndrome (OS) in Caucasians in Eastern Siberia. The sample size was 91 participants: the first group-30 patients with OS; the second group-30 patients AH; and the third group-31 healthy volunteers. RT-PCR was used for the determination of alleles and genotypes of the SNPs rs2779249 and rs2297518 of the NOS2 gene in all groups of participants. We showed that the frequency of allele A was significantly higher among patients with AH compared with healthy volunteers (p-value < 0.05). The frequency of the heterozygous genotype CA of rs2779249 was higher in the first group vs. the control (p-value = 0.03), and in the second group vs. the control (p-value = 0.045). The frequency of the heterozygous genotype GA of rs2297518 was higher in the first group vs. the control (p-value = 0.035), and in the second group vs. the control (p-value = 0.001). The allele A of rs2779249 was associated with OS (OR = 3.17 [95% CI: 1.31-7.67], p-value = 0.009) and AH (OR = 2.94 [95% CI: 1.21-7.15], p-value = 0.015) risks compared with the control. The minor allele A of rs2297518 was associated with OS (OR = 4.0 [95% CI: 0.96-16.61], p-value = 0.035) and AH (OR = 8.17 [95% CI: 2.03-32.79], p-value = 0.001) risks compared with the control. Therefore, our pilot study demonstrated that the SNPs rs2779249 and rs229718 of the NOS2 gene could be promising genetic biomarkers for this OS risk in Caucasians from Eastern Siberia.

Exposure to environmental air pollutants as a risk factor for primary Sjögren's syndrome.

Environmental etiology of primary Sjögren's syndrome (pSS), an autoimmune disease, has been proposed. This study determined whether the exposure to air pollutants was an independent risk factor for pSS. Participants were enrolled from a population-based cohort registry. Daily average concentrations of air pollutants from 2000 to 2011 were divided into 4 quartiles. Adjusted hazard ratios (aHRs) of pSS for exposure to air pollutants were estimated in a Cox proportional regression model adjusting for age, sex, socioeconomic status, and residential areas. A subgroup analysis stratified by sex was conducted to validate the findings. Windows of susceptibility indicated years of exposure which contributed the most to the observed association. Ingenuity Pathway Analysis was used to identify underlying pathways of air pollutant-associated pSS pathogenesis, using Z-score visualization. Two hundred patients among 177,307 participants developed pSS, with a mean age of 53.1 years at acumulative incidence of 0.11% from 2000 to 2011. Exposure to carbon monoxide (CO), nitric oxide (NO), and methane (CH4) was associated with a higher risk of pSS. Compared to those exposed to the lowest concentration level, the aHRs for pSS were 2.04 (95%CI=1.29-3.25), 1.86 (95%CI=1.22-2.85), and 2.21 (95%CI=1.47-3.31) for those exposed to high levels of CO, NO, and CH4, respectively. The findings persisted in the subgroup analysis, in which females exposed to high levels of CO, NO, and CH4 and males exposed to high levels of CO were associated with significantly great risk of pSS. The cumulative effect of air pollution on pSS was time-dependent. The underlying cellular mechanisms involved chronic inflammatory pathways including the interleukin-6 signaling pathway. Exposure to CO, NO, and CH4 was associated with a high risk of pSS, which was biologically plausible.

Kadar nitric oxide serum berhubungan dengan tekanan darah pada dewasa muda dengan obesitas sentral: studi pertama di Indonesia

Serum nitric oxide levels are associated with blood pressure in young adults with central obesityBackground: Central obesity is a risk factor for hypertension, which is closely related to the presence of endothelial dysfunction and associated with levels of nitric oxide (NO). Objective: This research was conducted to determine the relationship between NO and blood pressure (BP) in young adults with and without central obesity and also to compare the NO levels and BP between the two groups. Methods: This cross-sectional study was conducted in Pekanbaru, Riau, Indonesia, with 80 young adult subjects aged 18-25 years by consecutive sampling, consisting of 40 subjects with central obesity (waist circumference (WC) ≥90 cm and ≥80 cm for men and women, respectively) and 40 subjects without central obesity (WC &lt;90 cm and &lt; 80 cm for men and women, respectively). Blood pressure was measured using a digital sphygmomanometer, and NO levels were measured using Griess methods. The statistical analysis begins with the normality test of the data, normal data was analyzed with the Pearson correlation test, and abnormal data was analyzed with the Spearman test. Differences in the levels of NO, systolic blood pressure (TDS), and diastolic blood pressure (TDD) between groups was analyzed with the Mann-Whitney-U test.Results: Most of the subjects had high NO levels (66.3%). NO levels had a positively significant relationship with systolic blood pressure (SBP) and diastolic blood pressure (DBP) in total subjects (r = 0.503, p &lt;0.05; r= 0.289, p&lt;0.05, respectively) and with SBP in subjects with central obesity (r = 0.324, p &lt;0.05) but there was no significant relationship in normal subjects. There is a significant difference between serum NO levels, SBP, and DBP between subjects with central obesity and normal subjects (p &lt;0.05).Conclusion: NO levels have a positive significant relationship with SBP and DBP in total subjects and with SBP in young adults with central obesity.

Multifunctional antibiotics-free hydrogel dressings with self-regulated nitric oxide-releasing kinetics for improving open wound healing.

Healing of large open wounds remains a major challenge in clinics due to the high risk of bacterial infection and slow healing rates, while excessive use of antibiotics would cause enhanced antibiotic resistance and reduced biocompatibility. Herein, we developed a multifunctional hydrogel dressing (GCNO) through embedding nitrosothiol-conjugated chitosan into a crosslinked gelatin methacrylate (GelMA) network via hydrogen bonding, which presented self-regulated nitric oxide (NO) release kinetics for temporally controlled bacterial elimination and wound repair. At early stages after implantation, the positively charged chitosan molecules of the GCNO hydrogel precursors and release of a high level of NO from the GCNO hydrogel demonstrated effective coordinated antimicrobial capability, thus preventing wound infection in the early stages of healing. While at later stages of wound healing, the hydrogel could sustainably release low levels of NO to stimulate the proliferation and migration of fibroblasts and endothelial cells, leading to accelerated angiogenesis and cell deposition at the wound area. GCNO hydrogels exhibited anti-bacterial and wound repair performance with excellent biocompatibility and biosafety. Overall, this antibiotic-free GCNO hydrogel could demonstrate self-adaptive NO release profiles to prevent bacterial infection in early stages of wound healing while accelerating skin regeneration at later stages, which may offer new insights for the clinical management of large open wounds in clinics.

Relationship between histopathological lesions and oxidative stress in mice infected with Angiostrongylus costaricensis.

This study describes changes in oxidative stress (OS) parameters in mice experimentally infected with Angiostrongylus costaricensis, which causes abdominal angiostrongyliasis. For this, 28 Swiss mice were used, divided into two groups (G1 and G2), with 14 animals each. Of these, eight were infected with ten infective larvae each, by gavage, and six were used as a control group. Mice from G1 and G2 were euthanized at 14 days and 24 days post-infection, respectively. Tissue samples were used for histopathological analysis and blood (serum) samples were taken to assess the levels of proteins, non-protein thiols (NPTs) and nitric oxide (NO), from centrifugation and subsequent collection of aliquots of the supernatant. Among OS parameters, infected mice in both groups had higher NO levels than the control group, due to the presence of: eosinophil infiltrate in the liver and intestine; pancreatitis; and intestinal granuloma. However, the infected mice of both groups showed a reduction in the levels of NPTs, in relation to the control group, due to the presence of: eosinophilic infiltrate in the liver and intestine; and intestinal granuloma. Our results suggest that A. costaricensis infection has important effects on the intestine, liver and pancreas, and the analyses were performed from the tissue of these organs. The mechanisms for these changes are related to the decrease in the body's main antioxidant defences, as demonstrated by the reduction of NPTs, thus contributing to the development of more severe tissue damage. Thus, the objective of the present study was to evaluate the relationship between histopathological lesions and markers for OS.

Dengue induces iNOS expression and nitric oxide synthesis in platelets through IL-1R.

Dengue is an arthropod-born disease caused by dengue virus (DENV), that may manifest as a mild illness or severe form, characterized by hemorrhagic fever and shock. Nitric oxide (NO) is a vasodilator signaling molecule and an inhibitor of platelet aggregation known to be increased in platelets from dengue patients. However, the mechanisms underlying NO synthesis by platelets during dengue are not yet elucidated. IL-1β is a pro-inflammatory cytokine able to induce iNOS expression in leukocytes and present in dengue patients at high levels. Nevertheless, the role of IL-1β in platelet activation, especially regarding iNOS expression, are not clear. We prospectively followed a cohort of 28 dengue-infected patients to study NO synthesis in platelets and its relationship with disease outcomes. We used in vitro infection and stimulation models to gain insights on the mechanisms. We confirmed that platelets from dengue patients express iNOS and produce higher levels of NO during the acute phase compared to healthy volunteers, returning to normal levels after recovery. Platelet NO production during acute dengue infection was associated with the presence of warning signs, hypoalbuminemia and hemorrhagic manifestations, suggesting a role in dengue pathophysiology. By investigating the mechanisms, we evidenced increased iNOS expression in platelets stimulated with dengue patients´ plasma, indicating induction by circulating inflammatory mediators. We then investigated possible factors able to induce platelet iNOS expression and observed higher levels of IL-1β in plasma from patients with dengue, which were correlated with NO production by platelets. Since platelets can synthesize and respond to IL-1β, we investigated whether IL-1β induces iNOS expression and NO synthesis in platelets. We observed that recombinant human IL-1β enhanced iNOS expression and dose-dependently increased NO synthesis by platelets. Finally, platelet infection with DENV in vitro induced iNOS expression and NO production, besides the secretion of both IL-1α and IL-1β. Importantly, treatment with IL-1 receptor antagonist or a combination of anti-IL-1α and anti-IL-1β antibodies prevented DENV-induced iNOS expression and NO synthesis. Our data show that DENV induces iNOS expression and NO production in platelets through mechanisms depending on IL-1 receptor signaling.

Phytantriol-Based Berberine-Loaded Liquid Crystalline Nanoparticles Attenuate Inflammation and Oxidative Stress in Lipopolysaccharide-Induced RAW264.7 Macrophages

Inflammation and oxidative stress are interrelated processes that represent the underlying causes of several chronic inflammatory diseases that include asthma, cystic fibrosis, chronic obstructive pulmonary disease (COPD), allergies, diabetes, and cardiovascular diseases. Macrophages are key initiators of inflammatory processes in the body. When triggered by a stimulus such as bacterial lipopolysaccharides (LPS), these cells secrete inflammatory cytokines namely TNF-α that orchestrate the cellular inflammatory process. Simultaneously, pro-inflammatory stimuli induce the upregulation of inducible nitric oxide synthase (iNOS) which catalyzes the generation of high levels of nitric oxide (NO). This, together with high concentrations of reactive oxygen species (ROS) produced by macrophages, mediate oxidative stress which, in turn, exacerbates inflammation in a feedback loop, resulting in the pathogenesis of several chronic inflammatory diseases. Berberine is a phytochemical embedded with potent in vitro anti-inflammatory and antioxidant properties, whose therapeutic application is hindered by poor solubility and bioavailability. For this reason, large doses of berberine need to be administered to achieve the desired pharmacological effect, which may result in toxicity. Encapsulation of such a drug in liquid crystalline nanoparticles (LCNs) represents a viable strategy to overcome these limitations. We encapsulated berberine in phytantriol-based LCNs (BP-LCNs) and tested the antioxidant and anti-inflammatory activities of BP-LCNs in vitro on LPS-induced mouse RAW264.7 macrophages. BP-LCNs showed potent anti-inflammatory and antioxidant activities, with significant reduction in the gene expressions of TNF-α and iNOS, followed by concomitant reduction of ROS and NO production at a concentration of 2.5 µM, which is lower than the concentration of free berberine concentration required to achieve similar effects as reported elsewhere. Furthermore, we provide evidence for the suitability for BP-LCNs both as an antioxidant and as an anti-inflammatory agent with potential application in the therapy of chronic inflammatory diseases.

Ozone therapy in the treatment of the elderly

Aging is a universal process of constant reducing of the functioning of a complex system over time, affecting all levels of its organization that impede self-renewal processes. In the structure of the geroprotective therapeutic effect, the main role is given to combat oxidative stress. The most important natural remedy without drugs with this property is the ozoneoxygen mixture, whose effect on the human body is verified by studies conducted both nationally and internationally. 119 patients who underwent outpatient treatment at the Ozone Center in Nizhny Novgorod, Russia, were examined, the basic diagnosis was coronary heart disease, stable angina CF II-III. Parenteral administration of ozonated saline normalizes the pro- and antioxidant serum balance of elderly patients with coronary heart disease. After a course of ozone therapy, there is a decrease in the level of lipid metabolism - total cholesterol, LDL-C, triglycerides. Intravenous administration of low ozone levels improves endothelial function by lowering high levels of nitric oxide and endothelin. Ozone therapy reduces the ability of platelets to aggregate and improves the rheological properties of the blood of elderly patients with coronary heart disease.

Novel nitric oxide-releasing derivatives of pyranocarbazole as antitumor agents: Design, synthesis, biological evaluation, and nitric oxide release studies

In this study, a series of novel furoxan-based nitric oxide (NO) releasing derivatives of pyranocarbazole alkaloids were designed, synthesized, and biologically evaluated against human cancer cell lines. The derivatives showed considerable antiproliferative activities (IC50 = 0.05–7.55 μM) and most compounds showed higher activity in MDA-MB-231 than H460 and HeLa. Especially, the most active derivative 7a (IC50 = 0.05 μM) against MDA-MB-231 was about 60 times stronger than lead compound, as well as equivalent to positive control taxol, and produced high levels of NO in MDA-MB-231. Furthermore, 7a could significantly inhibit the growth of MDA-MB-231 tumors in vivo with low toxicity and the PI3K/Akt signaling pathway. These results indicated that compound 7a could be a promising lead for further studies.

Effects of dulaglutide on endothelial progenitor cells and arterial elasticity in patients with type 2 diabetes mellitus

BackgroundRandomised controlled trial showed that dulaglutide can reduce the risk of atherosclerotic cardiovascular disease (ASCVD) in patients with type 2 diabetes mellitus (T2DM), but the underlying mechanisms remain unclear. This study aimed to investigate the effect of dulaglutide on the number and function of endothelial progenitor cells (EPCs) in the peripheral blood of patients with T2DM and its role in improving arterial elasticity, so as to determine potential mechanisms of preventive effect of dulaglutide on ASCVD.MethodsSixty patients with T2DM were treated with 1000 mg/day of metformin and randomly divided into two groups for 12 weeks: metformin monotherapy group (MET group, n = 30), and metformin combined with dulaglutide group (MET-DUL group, n = 30). Before and after treatment, the number of CD34+CD133+KDR+ EPCs and the brachial–ankle pulse wave velocity (baPWV) of the participants were measured, and EPC proliferation, adhesion, migration, and tubule formation were assessed in vitro.ResultsThere were no significant differences in the number and function of EPCs and baPWV changes in MET group (P > 0.05). In MET-DUL group, nitric oxide (NO) levels and the number of EPCs increased after treatment (P < 0.05), while the levels of C-reactive protein (CRP), interleukin-6 (IL-6), tumour necrosis factor-α (TNF-α), advanced glycation end products (AGEs), and baPWV decreased (P < 0.05). EPC proliferation, adhesion, migration, and tubule formation abilities were significantly enhanced (P < 0.05). Correlation analysis showed that in MET-DUL group, the changes in CRP, IL-6, TNF-α, and AGEs were negatively correlated with the number of EPCs and their proliferation and migration abilities (P < 0.05). Body weight, NO, CRP, and IL-6 levels were independent factors affecting the number of EPCs (P < 0.05). The changes in number of EPCs, proliferation and migration abilities of EPCs, and NO and IL-6 levels were independent influencing factors of baPWV changes (P < 0.05).ConclusionDulaglutide can increase the number and function of EPCs in peripheral blood and improve arterial elasticity in patients with T2DM; it is accompanied by weight loss, inflammation reduction, and high NO levels. Dulaglutide regulation of EPCs may be a mechanism of cardiovascular protection.

Effect of zinc oxide nanoparticles on cellular stress in Ehrlich ascites carcinoma

Background Cancer cells are characterized by high cellular stress that is considered crucial for cancer cell survival and metastasis. Aim In this study, we assessed cellular stress in Ehrlich ascites carcinoma (EAC) cells before and after treatment with intraperitoneal zinc oxide nanoparticles. Materials and methods This experiment was performed on six groups of mice: 90 female albino mice were included in this study and were divided into six groups (n=15 per group): C [saline treated mice (20 ml/Kg intraperitoneal (i.p.)], CZn20/2d [ZnO NP-treated mice (20 mg/kg i.p. every other day], EC [untreated EAC-bearing mice], EZn 10/d [EAC-bearing mice administered with ZnO NPs 10 mg/kg everyday], EZn 10/2d [EAC-bearing mice administered with ZnO NPs 10 mg/kg every other day], EZn20/2d [EACs-bearing mice administered with ZnO NPs 20 mg/kg every other day]. After 2 weeks, the ascitic fluid and mammary gland were collected to measure the level of nitric oxide (NO) and peroxynitrite. Results Based on this results, there was a significantly high level of NO and peroxynitrite in the EAC groups compared with the control group. Zinc oxide nanoparticles significantly decreased the level of NO and peroxynitrite in the EAC groups compared with the control group. Conclusion This study implied that cancer cells have higher cellular stress, and it was efficiently decreased by treatment with zinc oxide nanoparticles indicating its efficacy against cancer cells.