Diclofenac exhibits cytotoxic activity associated with metabolic alterations and p53 induction in ESCC cell lines and decreases ESCC tumor burden in vivo.

Esophageal squamous cell carcinoma (ESCC) is one of the most aggressive forms of human malignancy, often displaying limited therapeutic response. Here, we examine the non-steroidal anti-inflammatory drug diclofenac (DCF) as a novel therapeutic agent in ESCC using complementary in vitro and in vivo models. DCF selectively reduced viability of human ESCC cell lines TE11, KYSE150, and KYSE410 as compared with normal primary or immortalized esophageal keratinocytes. Apoptosis and altered cell cycle profiles were documented in DCF-treated TE11 and KYSE 150. In DCF-treated TE11, RNA-Sequencing identified differentially expressed genes and Ingenuity Pathway Analysis predicted alterations in pathways associated with cellular metabolism and p53 signaling. Downregulation of proteins associated with glycolysis was documented in DCF-treated TE11 and KYSE150. In response to DCF, TE11 cells further displayed reduced levels of ATP, pyruvate, and lactate. Evidence of mitochondrial depolarization and superoxide production was induced by DCF in TE11 and KYSE150. In DCF-treated TE11, the superoxide scavenger MitoTempo improved viability, supporting a role for mitochondrial reactive oxygen species in DCF-mediated toxicity. DCF treatment resulted in increased expression of p53 in TE11 and KYSE150. p53 was further identified as a mediator of DCF-mediated toxicity in TE11 as genetic depletion of p53 partially limited apoptosis in response to DCF. Consistent with the anticancer activity of DCF in vitro, the drug significantly decreased tumor burdene in syngeneic ESCC xenograft tumors and 4-nitroquinoline 1-oxide-mediated ESCC lesions in vivo. These preclinical findings identify DCF as an experimental therapeutic that should be explored further in ESCC.

Hesperidin alleviates vascular dysfunction and remodelling in high-fat/high-fructose diet-fed rats by modulating oxidative stress, inflammation, AdipoR1, and eNOS expression.

Hesperidin, a flavanone glycoside, has shown antihypertensive, antioxidant, and anti-inflammatory effects. In the present study, the therapeutic effects of hesperidin on vascular function and remodelling, and possible underlying mechanisms involved in high-fat/high-fructose diet (HFFD)-fed rats were investigated. Male Sprague-Dawley rats were fed a high-fat diet plus 15% fructose in drinking water for 16 weeks. HFFD-fed rats were treated with hesperidin (30 mg/kg/day) or vehicle for the last 4 weeks. Treatment of HFFD-fed rats with hesperidin significantly attenuated metabolic alterations, vascular endothelial dysfunction and remodelling. Hesperidin markedly alleviated HFFD-induced oxidative stress and inflammation by decreasing plasma malondialdehyde level and aortic superoxide anion production, and by suppressing aortic tumor necrosis factor-α and interleukin-6 overexpression. Additionally, increased plasma levels of the adiponectin and nitric oxide metabolite, together with restoration of adiponectin receptor 1 (AdipoR1) and endothelial nitric oxide synthase (eNOS) protein expression, were also observed after treatment with hesperidin. These findings indicated that hesperidin alleviates HFFD-induced vascular dysfunction and remodelling in rats. The possible underlying mechanism may involve the reduction of oxidative stress and inflammation, and the restoration of AdipoR1and eNOS expression.

Persistent CD38 Expression on CD8 + T Lymphocytes Contributes to Altered Mitochondrial Function and Chronic Inflammation in People With HIV, Despite ART.

BACKGROUND: Age-associated comorbidities are higher in people with HIV (PWH) than HIV-negative individuals. This is partially attributed to immune activation and CD38 expression on T cells driving chronic inflammation. However, the exact contribution of CD38-expressing T cells on the proinflammatory response is not completely understood. METHODS: CD38-expressing CD8 + T lymphocytes were measured from PWH and HIV-negative individuals. Mitochondrial mass, superoxide content, membrane depolarization of CD4 + and CD8 + T lymphocytes, and cytokine production after HIV(Gag)-specific peptide stimulation from CD38 + CD8 + T lymphocytes of PWH were measured to link biological effects of CD38 expression on cellular metabolism. RESULTS: The frequency of activated CD8 + CD38 + T cells persists in PWH on ART compared with HIV-negative individuals. Higher CD38 expression is associated with mitochondrial biogenesis and HIV(Gag)-specific proinflammatory cytokine production in PWH. Blockade of CD38 results in lower Gag-specific cytokine production. CONCLUSIONS: ART only partially reduced HIV-induced CD38 expression on CD8 + T cells. CD8 + CD38 + T cells are highly activated in vivo, and HIV-specific stimulation in vitro augments CD38 expression, contributing to a proinflammatory response despite virologic control with ART. Therefore, CD38 is a potential therapeutic target for mitigating chronic inflammation that likely drives cellular aging, comorbidities, and end-organ disease in PWH.

Plumbagin Enhances the Anticancer Efficacy of Cisplatin by Increasing Intracellular ROS in Human Tongue Squamous Cell Carcinoma.

Cisplatin is widely used in the treatment of tongue squamous cell carcinoma (TSCC), but its clinical efficacy is limited by drug resistance and toxic side effects. Hence, a novel compound capable of enhancing the anticancer effect of cisplatin while reducing the side effects is urgently needed. We have previously shown that plumbagin (PLB), an anticancer phytochemical, is able to inhibit the growth of TSCC in vitro and in vivo. The objective of this study was to investigate the effect of PLB in reversing the resistance of TSCC to cisplatin as well as its molecular mechanisms. Here, we found that PLB enhances cisplatin-induced cytotoxicity, apoptosis, and autophagy in CAL27 and cisplatin-resistant CAL27/CDDP cells. PLB could inhibit the viability and growth of TSCC cells by increasing the production of intracellular reactive oxygen species (ROS). In addition, the combination treatment of PLB and cisplatin resulted in a synergistic inhibition of TSCC viability, apoptosis, and autophagy by increasing intracellular ROS, which may be achieved by activating JNK and inhibiting AKT/mTOR signaling pathways. Finally, the synergistic treatment was also demonstrated in vivo. Therefore, PLB combined with cisplatin is a potential therapeutic strategy against therapy TSCC cisplatin resistance.

The Use of a Novel Super-Oxidized Solution on Top of Standard Treatment in the Home Care Management of Postsurgical Lesions of the Diabetic Foot Reduces Reinfections and Shortens Healing Time.

There are still many unmet needs in the treatment of chronic wounds, especially regarding microenvironment modulation. Nexodyn is a novel super-oxidized solution capable of contrast bacterial proliferation. We aimed to evaluate if this solution, on top of standard of care, was safe and effective in improving diabetic foot outcome. We selected 50 patients admitted in our department to be submitted to surgery for acute diabetic foot infection. All patients were left open to heal for secondary intent. We divided patients into 2 groups: Group A (n = 25, male/female = 17/8, age = 67.3 ± 12.1 years, hemoglobin A1C = 7.9 ± 1.1%), patients treated with standard of care and, on top of this, Nexodyn solution, and Group B, treated only with standard of care. After discharge, patients continued Nexodyn application. We followed up patients until complete healing or up to 6 months. No differences between groups in healing rate, while time required for complete healing was significantly shorter in Group A (64.9 ± 12.1 days vs 147.4 ± 23.1 days, P < .01). During follow-up, the group treated with Nexodyn showed a reduced rate of reinfections (12 patients in Group B vs 3 patients in Group A, P < .05) and of further debridement procedures (1 patient in Group A vs 10 patients in Group B, P < .05). Nexodyn provided effective protection against reinfections in diabetic foot patients, thus reducing the necessity for debridement procedures and their healing time and presents a safety profile similar to saline solution.

[Oxidative stress and natural antioxidant mechanisms: the role in neurodegeneration. From molecular mechanisms to therapeutic strategies]. / Stres oksydacyjny i naturalne mechanizmy antyoksydacyjne ­ znaczenie w procesie neurodegeneracji. Od mechanizmów molekularnych do strategii terapeutycznych.

A lot of evidence exists that oxidative stress is the primary cause of neurodegeneration. Neurons are more susceptible to oxidative damage than other cells due to their high oxygen consumption, low activity of antioxidant enzymes, elevated concentration of polyunsaturated fatty acids in the cell membrane, high number of mitochondria, unfavorable space/volume ratio and vicinity of microglia cells which are likely to produce increased amounts of superoxide radical. Moreover, the tendency to accumulate transition metals in the brain creates a higher probability of Fenton's reaction occurring, a product of which is a hydroxyl radical. Lower activities of natural antioxidants as well as higher concentrations of markers of oxidative damage to proteins, lipids and DNA were observed in patients with neurodegenerative diseases in relation to healthy individuals. There is a lot of research being conducted to develop effective and safe antioxidants that would be useful in the therapy or prevention of neurodegenerative diseases.  

Efficacy and safety of oxum in treatment of the venous ulcer.

The present study was carried out to find out the efficacy and safety of oxum in the treatment of venous ulcers. The oxum (superoxidised water) is a pH neutral, non-irritating, aqueous solution that possesses a good antiseptic, antimicrobial activity and wound healing properties. The study was conducted in 30 patients of venous ulcers with a culture examination positive for pathogenic microbial flora. All patients received a gauze dressing impregnated with oxum followed by compression bandage for 28 days. The primary endpoint was the calculation of ulcer size using ulcer tracing. Assessment of periwound oedema, periwound erythema, wound fibrin and wound granulations were considered as secondary endpoints. There was a singificant reduction in ulcer size starting from day 7 of the treatment. Significant improvements in secondary endpoints were observed. This study has demonstrated that oxum improved the clinical status, reduced the signs of inflammation in venous ulcers in addition to its well confirmed anti-infective properties.

Efficacy and safety of neutral pH superoxidised solution in severe diabetic foot infections.

The use of antiseptics in wound care is often controversial and there is definitely a need for a non toxic, highly disinfective agent. This study assessed the efficacy of a neutral pH superoxidised aqueous solution (NpHSS) for infection control, odour reduction and surrounding skin and tissue damage on infected diabetic foot ulcerations. From November 2003 to March 2004, 45 patients with type 2 diabetes were randomised into a single-blind clinical trial comparing NpHSS (intervention group; n = 21) versus conventional disinfectant (control group; n = 16). All patients received comprehensive care including surgical debridement as appropriate, moist wound care, intensive glucose control and broad spectrum antibiotics. Treatment groups were matched in terms of sex, age (61.9 +/- 11.9 versus 67.8 +/- 11.6), years of diabetes duration (16.4 +/- 8.1 versus 17 +/- 10.2), obesity, HgAlc (7.1 +/- 2 versus 6.7 +/- 1.8), initial fasting glycaemia (163 +/- 59 versus 152 +/- 65.8 mg/dl), ulcer duration/week (13.7 +/- 24 versus 15.1 +/- 16.3), B/A Index (0.9 +/- 0.5 versus 1.14 +/- 0.7), depth and extent of infection/periwound cellulitis (groups B and C of the Tampico Hospital Classification) as well as aetiology (P = 0.647). Odour reduction was achieved in all NpHSS patients (100% versus 25%; P < 0.01) and surrounding cellulitis diminished (P < 0.001) in 17 patients (80.9% versus 43.7%). Nineteen patients in the NpHSS group showed advancement to granulating tissue stage (90.4% versus 62.5%; P = 0.05) with significantly less tissue toxicity (94% versus 31.2%; P < 0.01). A non toxic, NpHSS, as part of a comprehensive care regimen, may be more efficacious in infection control, odour and erythema reduction than conventional disinfectants in treatment of diabetic foot infections.

Reactive oxygen species as essential components of ambient air.

In this review evidence for the presence of the anion radical O2(-*) in atmospheric air is considered, and the biological activity of superoxide and negative air ions is compared. Various aspects of the biological effect of superoxide and other reactive oxygen species contained in air at the cell, tissue, and organism levels are discussed. The results of the therapeutic use of exogenous gaseous superoxide and low doses of H2O2 for the treatment of bronchial asthma, pain, and Parkinson's disease are reported. A hypothesis on the mechanism of physiological action of exogenous reactive oxygen species is discussed.

Role of nitric oxide and superoxide in Giardia lamblia killing.

Giardia lamblia trophozoites were incubated for 2 h with activated murine macrophages, nitric oxide (NO) donors or a superoxide anion generator (20 mU/ml xanthine oxidase plus 1 mM xanthine). Activated macrophages were cytotoxic to Giardia trophozoites (approximately 60% dead trophozoites). The effect was inhibited (> 90%) by an NO synthase inhibitor (200 microM) and unaffected by superoxide dismutase (SOD, 300 U/ml). Giardia trophozoites were killed by the NO donors, S-nitroso-acetyl-penicillamine (SNAP) and sodium nitroprusside (SNP) in a dose-dependent manner (LD50 300 and 50 microM, respectively). A dual NO-superoxide anion donor, 3-morpholino-sydnonimine hydrochloride (SIN-1), did not have a killing effect in concentrations up to 1 mM. However, when SOD (300 U/ml) was added simultaneously with SIN-1 to Giardia, a significant trophozoite-killing effect was observed (approximately 35% dead trophozoites at 1 mM). The mixtures of SNAP or SNP with superoxide anion, which yields peroxynitrite, abolished the trophozoite killing induced by NO donors. Authentic peroxynitrite only killed trophozoites at very high concentrations (3 mM). These results indicate that NO accounts for Giardia trophozoites killing and this effect is not mediated by peroxynitrite.

[Superoxide inhalation as adjuvant therapy in children with bronchial asthma]. / Ingaliatsiia superoksida kak ad''iuvantnyi metod lecheniia detei, bol'nykh bronchial'noi astmoi.

The aim of the present investigation was to confirm results of an earlier pilot study on the adjuvant treatment of atopic asthmatics using superoxide inhalation in a double blind placebo controlled experiment. Group 1 (verum) included 28 patients (7 girls and 21 boys whose age was 12.2c2.9 years) and Group 2 (placebo) comprised 23 (8 girls and 15 boys aged 11.7c3.2 years). Fifteen-minute superoxide inhalations were used as adjuvant therapy to the routine treatment. The patients underwent spirometry, blood analysis, nasal smear tests for eosinophils, and questionnaires. The worse the baseline parameters were, the better their spirometric ones were. This was reflected by higher significant correlations between the baseline values and their changes in peak flowmetry, FEV1, and FEF and by the patients' grouping by the baseline limitation of respiratory functions (less than 80% or more of the individual normal values). The most significant improvement (46.5%; p = 0.008) was observed in the subgroup with VC. In accordance with subjective assessment data, the time course of reductions in the frequency and severity of asthma attacks and coughing fits, and sputum amount was significantly better in Group 1 than in Group 2. The findings are interpreted as a result of an alleviated inflammatory process due to superoxide inhalation-induced oxidative training in patients.

Free radical-scavenging activity of Crassostera gigas extract (JCOE).

In the present study, we evaluate the free radical-scavenging activity of JCOE (Japan clinic oyster extract), a powder extracted from Crassostera gigas by a spin-trapping method using electron paramagnetic resonance (EPR), and also estimate the protective effect against gastric mucosal cell injury induced by hydrogen peroxide. The EPR study demonstrated that JCOE directly scavenged superoxide radical as well as hydroxyl radical in a concentration-dependent manner. After exposure to hydrogen peroxide for 4 h in Hank's balanced buffered solution, cell viability of rat gastric mucosal cells (RGM-1) was measured by modified MTT assay. Hydrogen peroxide-induced injury was not reversed by 1-h preincubation with 100 to 1,000 micrograms/mL JCOE solution which has high reactivity to hydroxyl radicals, indicating that the active ingredients, including taurine of JCOE on scavenging action of hydroxyl radical, did not penetrate cell membranes easily. Twenty-four hour pretreatment with the JCOE solution significantly reversed the decrease in cell viability induced by hydrogen peroxide, indicating the possibility that JCOE solution may stimulate the endogenous eliminating system against hydrogen peroxide.

Role of nitric oxide and superoxide in Giardia lamblia killing

Giardia lamblia trophozoites were incubated for 2 h with activated murine macrophages, nitric oxide (NO) donors or a superoxide anion generator (20 mU/ml xanthine oxidase plus 1 mM xanthine). Activated macrophages were cytotoxic to Giardia trophozoites (~60 per cent dead trophozoites). This effect was inhibited (>90 per cent) by an NO synthase inhibitor (200 muM) and unaffected by superoxide dismutase (SOD, 300 U/ml). Giardia trophozoites were killed by the NO donors S-nitroso-acetyl-penicillamine(SNAP)and sodium nitroprusside (SNP) in a dose-dependent manner (LD50 300 and 50 muM, respectively). A dual NO-superoxide anion donor, 3-morpholino-sydnonimine hydrochloride (SIN-1), did not have a killing effect in concentration up to 1 mM. However, when SOD (300 U/ml) was added simultaneously with SIN-1 to Giardia, a significant trophozoite-killing effect was observed (~35 per cent dead trophozoites at 1 mM). The mixture of SNAP or SNP with superoxide anion, which yields peroxynitrite, abolished the trophozoite killing induced by NO donors. Authentic peroxynitrite only killed trophozoites at very high concentrations (3 mM). These results indicate that NO accounts for Giardia trophozoite killing and this effect is not mediated by peroxynitrite.

What should an ideal antioxidant do (and not do)?

Superoxide, H2O2 and particularly activated hydroxyl radicals (OH.) can cause lipid peroxidation, destroy enzymes and anti-elastases, produce DNA breaks and lead to death of mammalian cells. Conversely, these active oxygen species may also be very important for the killing of some microbial infective agents. Clearly the therapeutic potential for reducing the destructive activity of oxidants must be offset by any harm which may result from inhibition of these protective actions of oxidants. The challenge lies in providing the one without the other.

Effect of 24-hour preservation with oxygen free radical scavengers on isolated-perfused canine heart-lungs.

The effectiveness of 24-hour hypothermic machine perfusion with TP-V (a hyperosmolar colloid solution containing dextrose, sucrose and ATP-MgCl2) alone, or in combination with oxygen free radical scavengers, was evaluated in isolated-perfused canine heart-lungs. Heart-lungs were perfused at 4 degrees C in either TP-V (n = 6), TP-V/Allopurinol (500 mg/L) (n = 6), or TP-V/Allopurinol (500 mg/L) & Catalase (5000 U/L) (n = 5). Lung inflation was maintained with 100% nitrogen. Following preservation, the heart-lungs were perfused with an albumin-mannitol perfusate for 3 hours at 37 degrees C, for functional, hemodynamic, and laboratory determinations. Cold preservation with TP-V/Allopurinol, and TP-V/Allopurinol & Catalase resulted in physiologically normal LDH levels during the 3-hour normothermic isolated perfusion test period. Significantly lower enzyme activity for CPK was evident at 0 (p less than .005) and 3 hours (p less than .05) of perfusion, while no significant differences in lactate production were seen among the groups. In addition, pH, PCO2, PO2, and left ventricular, aortic, and coronary artery pressures all remained within normal physiologic range, with no significant differences seen among the three groups. 99m Technetium scans demonstrated adequate patency among the heart-lungs, with better flow seen in those perfused with TP-V/Allopurinol & Catalase. Histological specimens confirmed a decrease in myocardial and pulmonary damage when Allopurinol and/or Catalase was used. It appears that oxygen free radical scavengers provide some protection from canine heart-lungs which have been hypothermically preserved for 24 hours.

Effect of 24 hour preservation with oxygen free radical scavengers on isolated perfused canine heart lungs

Los efectos de TP-V, una solución hiperosmolar con dextrosa, sucrosa, y ATP-MgCl2 fueron estudiados cuando usada sola o en combinación con substancias que eliminan los radicales libres de oxigenio, por 24 horas de perfusión hipotermica pulsátil de corazón pulmón y después revalorados en un sistema aislado de corazón pulmón a 37-C. Tres grupos de corazón pulmón de perros se analizaron: 1) TP-V (n=6), 2) TP-V con alopurinol (500mg/L) (m=6), 3) TP-V con alopurinol (500mg/L) y catalase (5000U/L) (n=5). Después de 24 horas de preservación, la preparación corazón pulmón fue perfundida con albúmina-manitol por 3 horas a 37-C con objeto de estimar su capacidad funcional y hemodinámica. Grupos 2 y 3 mostraron mejores valores de LDH y CPK a 0 t 3 horas de perfusión. Los valores de pH, pCO2, también como la presión ventricular izquierda, coronaria y aórtica, no cambinaron en forma significante. Histologicamente, el grupo 3 mostró mejor preservación del corazón y pulmones. En resúmen, podemos indicar que la solución TP-V, con las substancias que eliminan radicales libres de oxigeno, ofrecen una buena protección para la preparación corazón pulmón almacenada por 24 horas bajo hipotermia

Superoxide and inflammation: a mechanism for the anti-inflammatory activity of superoxide dismutase.

Several models of inflammation induced in laboratory animals were markedly inhibited by intravenous administration of the enzyme superoxide dismutase. In most cases, the enzyme required chemical modification to increase its circulating lifetime. In all cases, histological examination of the sites of the potential lesions revealed that treatment with superoxide dismutase prevented the accumulation of inflammatory cells. Experiments in vitro demonstrated the existence of a plasma factor which reacts with superoxide to become a potent chemotactic factor for neutrophils. The factor apparently serves to call increasing numbers of neutrophils to the inflammatory locus in response to the superoxide produced by the first cells to encounter the stimulus.