Reactive Oxygen Species/reactive Nitrogen Species Research Articles

Effects of prolonged application of peracetic acid-based disinfectant on recirculating aquaculture systems stocked with Atlantic salmon parr

The use of recirculating aquaculture systems (RAS) for Atlantic salmon (Salmo salar) production has become increasingly common. RAS water disinfection plays a crucial role on its biosecurity. Peracetic acid (PAA) is a promising disinfectant due to its powerful oxidative properties, broad antimicrobial spectrum, and rapid degradation into no harmful compounds. This study focused on assessing the consequences of prolonged application of a PAA-based disinfectant in a RAS stocked with salmon parr. The experiment included three treatment groups in triplicate: 0 mg/L PAA (control), 0.1 mg/L PAA, and 1 mg/L PAA, using nine-replicated RAS with a total of 360 fish (14.8 ± 2.3 g; N = 40/RAS). The study spanned 28 days, with samples collected on days 0, 14, and 28. The analyzed parameters were water quality, and fish parameters, including external welfare indicators, gill histology, total antioxidant capacity (TAC), reactive oxygen species/reactive nitrogen species (ROC/RNC), oxidative stress biomarkers related to DNA and protein, cellular DNA damage, and global gene expression. While water quality remained relatively stable, there was an increase in bacterial populations in the groups exposed to PAA, particularly 1 mg/L PAA. Fish weight did not differ between the control and PAA-exposed groups. TAC, ROC/RNC, and oxidative stress biomarkers exhibited similar trends. The study identified >400 differentially expressed genes (DEGs) in the skin, gill, and olfactory organ, with many of these DEGs associated with immune responses. Comparing the transcriptomic profiles of the three tissue organs revealed that the olfactory organ was the most reactive to PAA treatment. This study shows that calculated PAA concentrations of 0.1 mg/L and 1 mg/L in the pump-sump, contributed to an increase of bacteria whereas no detectable differences in health and welfare of salmon parr were found. These findings are promising for the implementation of PAA-based disinfectants in RAS stoked with Atlantic salmon parr.

Clinical and Biochemical Effects of Antioxidant Gel as a Local Drug Delivery Agent in Stage II Grade A Periodontitis Patients: A Prospective Clinical Study.

Background Periodontal tissue breakdown is mainly due to pathogenic bacteria and dysregulated immune response resulting in the production of reactive oxygen species/reactive nitrogen species (ROS/RNS) causing tissue degradation. Scaling and root planing (SRP)isusually donefor the management of periodontitis. However,it has been reported thatadjuncts like antibiotics, antiseptics, and antioxidants in the form of local drug delivery enhance the outcome of SRP. Aim The present clinical study aims toexamine the efficacy of an antioxidant oral gel (Bluem®) as a local drug delivery agent adjunct to SRPin the management of stage II grade A periodontitis in terms of clinical and biochemical parameters. Materials and methods The prospective clinical study was conducted among 40 stage II grade A periodontitis patients. The participants were then divided into two groups: Group 1 (Control)-SRP alone (n=20)and Group 2 (Test)-antioxidant gel (Bluem®) with SRP (n=20). Clinical variables including plaque index (PI), gingival index (GI), probing depth (PD), and clinical attachment level (CAL) were recorded. Saliva (unstimulated) specimens were collected to measure total oxidant status (TOS), total antioxidant capacity (TAOC), and oxidative stress index (OSI). Specimen collection and assessment of clinical variables were done before intervention (baseline) and after three months. SPSS Software (Version 20.0, Armonk, NY, USA: IBM Corp) was used for statistical analysis. Intragroup and intergroup comparisons were done by paired t-test and independent t-test, respectively. A p-value <0.05 indicated that the result was statistically significant. Results On intragroup analysis, both the groups at three months revealed statistically significant improvement of PI, GI, PD, CAL, TOS, TAOC, and OSI (p<0.05) from baseline. Intergroup comparison in the third month showed a statistically significant improvement in favor of Group 2 in terms of all the clinical and biochemical parameters (p<0.05) except for PI (p>0.05). Conclusion The locally delivered antioxidant gel as an adjunct to SRP seemsto be effective in reducing oxidative stress and improving the periodontal parameters among stage II grade A periodontitis patients.

Melatonin/Sericin Wound Healing Patches: Implications for Melanoma Therapy.

Melatonin and sericin exhibit antioxidant properties and may be useful in topical wound healing patches by maintaining redox balance, cell integrity, and regulating the inflammatory response. In human skin, melatonin suppresses damage caused by ultraviolet radiation (UVR) which involves numerous mechanisms associated with reactive oxygen species/reactive nitrogen species (ROS/RNS) generation and enhancing apoptosis. Sericin is a protein mainly composed of glycine, serine, aspartic acid, and threonine amino acids removed from the silkworm cocoon (particularly Bombyx mori and other species). It is of interest because of its biodegradability, anti-oxidative, and anti-bacterial properties. Sericin inhibits tyrosinase activity and promotes cell proliferation that can be supportive and useful in melanoma treatment. In recent years, wound healing patches containing sericin and melatonin individually have attracted significant attention by the scientific community. In this review, we summarize the state of innovation of such patches during 2021-2023. To date, melatonin/sericin-polymer patches for application in post-operational wound healing treatment has been only sparingly investigated and it is an imperative to consider these materials as a promising approach targeting for skin tissue engineering or regenerative dermatology.

Oxidative versus Reductive Stress in Breast Cancer Development and Cellular Mechanism of Alleviation: A Current Perspective with Anti-breast Cancer Drug Resistance.

Redox homeostasis is essential for keeping our bodies healthy, but it also helps breast cancer cells grow, stay alive, and resist treatment. Changes in the redox balance and problems with redox signaling can make breast cancer cells grow and spread and make them resistant to chemotherapy and radiation therapy. Reactive oxygen species/reactive nitrogen species (ROS/RNS) generation and the oxidant defense system are out of equilibrium, which causes oxidative stress. Many studies have shown that oxidative stress can affect the start and spread of cancer by interfering with redox (reduction-oxidation) signaling and damaging molecules. The oxidation of invariant cysteine residues in FNIP1 is reversed by reductive stress, which is brought on by protracted antioxidant signaling or mitochondrial inactivity. This permits CUL2FEM1B to recognize its intended target. After the proteasome breaks down FNIP1, mitochondrial function is restored to keep redox balance and cell integrity. Reductive stress is caused by unchecked amplification of antioxidant signaling, and changes in metabolic pathways are a big part of breast tumors' growth. Also, redox reactions make pathways like PI3K, PKC, and protein kinases of the MAPK cascade work better. Kinases and phosphatases control the phosphorylation status of transcription factors like APE1/Ref-1, HIF-1, AP-1, Nrf2, NF-B, p53, FOXO, STAT, and - catenin. Also, how well anti-breast cancer drugs, especially those that cause cytotoxicity by making ROS, treat patients depends on how well the elements that support a cell's redox environment work together. Even though chemotherapy aims to kill cancer cells, which it does by making ROS, this can lead to drug resistance in the long run. The development of novel therapeutic approaches for treating breast cancer will be facilitated by a better understanding of the reductive stress and metabolic pathways in tumor microenvironments.

Persistent Organic Pollutants released from decomposed adipose tissue affect mitochondrial enzyme function in the brain and eyes other than the liver.

Persistent organic pollutants (POPs) are toxic chemicals that can accumulate in the human body, and particularly in adipose tissue. POPs can induce metabolic diseases via mitochondrial dysfunction and can also cause cancer, obesity, and cardiovascular and neurodegenerative diseases. Although the effects of POPs were studied by evaluating mitochondrial function, which is fundamental in investigating the etiologies of various metabolic diseases, the physiological impact of POPs released by the decomposition of fat in adipose tissue is barely understood. Therefore, to investigate the mitochondrial dysfunction caused by POPs released from adipose tissue to other organs, zebrafish were exposed to POPs and placed into four groups: control (C), obesity control (OC), obesity control with POPs (OP), and POP exposure with obesity and caloric restriction (OPR). Next, the activities of the mitochondrial respiratory complexes and the levels of ATP production, reactive oxygen species/reactive nitrogen species (ROS/RNS), and antioxidants, such as glutathione and superoxide dismutase, were measured in the brain, eyes, and liver, as these are the major organs most susceptible to metabolic diseases. POPs released from adipose tissue showed a stronger effect than the direct effects of obesity and POPs on mitochondrial enzyme activity in the brain and eye. Released POPs increased mitochondrial complex I activity and decreased mitochondrial complex II activity compared with normal, obesity, and POP-treated conditions in the brain and eyes. However, the mitochondrial complexes' activities in the liver were affected more by obesity and POPs. In the liver, the mitochondrial enzyme activities of the OPR group seemed to recover to the control level, but it was slightly lowered in the OC and OP groups. Independently, the ROS/RNS and antioxidant levels were not affected by obesity, POPs, or the released POPs in the brain, eye, and liver. The results indicate that POPs stored in adipose tissue and released during fat decomposition did not affect oxidative stress but could affect mitochondrial respiratory enzymes in organ dependent manner. This study is meaningful in that it provides experimental evidence that stored POPs affect specific organs for prolonged periods and can be linked to various diseases in advance.

Detection of micro-plasma-induced exosomes secretion in a fibroblast-melanoma co-culture model

BackgroundMelanoma is a highly aggressive tumor and a significant cause of skin cancer-related death. Timely diagnosis and treatment require identification of specific biomarkers in exosomes secreted by melanoma cells. In this study, label-free surface-enhanced Raman spectroscopy (SERS) method with size-matched selectivity was used to detect membrane proteins in exosomes released from a stimulated environment of fibroblasts (L929) co-cultured with melanoma cells (B16–F10). To promote normal secretion of exosomes, micro-plasma treatment was used to gently induce the co-cultured cells and slightly increase the stress level around the cells for subsequent detection using the SERS method. Results and discussionFirstly, changes in reactive oxygen species/reactive nitrogen species (ROS/RNS) concentrations in the cellular microenvironment and the viability and proliferation of healthy cells are assessed. Results showed that micro-plasma treatment increased extracellular ROS/RNS levels while modestly reducing cell proliferation without significantly affecting cell survival. Secondly, the particle size of secreted exosomes isolated from the culture medium of L929, B16–F10, and co-cultured cells with different micro-plasma treatment time did not increase significantly under single-cell conditions at short treatment time but might be changed under co-culture condition or longer treatment time. Third, for SERS signals related to membrane protein biomarkers, exosome markers CD9, CD63, and CD81 can be assigned to significant Raman shifts in the range of 943–1030 and 1304-1561 cm-1, while the characteristics SERS peaks of L929 and B16–F10 cells are most likely located at 1394/1404, 1271 and 1592 cm-1 respectively. Significance and noveltyTherefore, this micro-plasma-induced co-culture model provides a promising preclinical approach to understand the diagnostic potential of exosomes secreted by cutaneous melanoma/fibroblasts. Furthermore, the label-free SERS method with size-matched selectivity provides a novel approach to screen biomarkers in exosomes secreted by melanoma cells, aiming to reduce the use of labeling reagents and the processing time traditionally required.

Analysis of In Vitro Leukocyte Responses to Biomaterials in the Presence of Antimicrobial Porcine Neutrophil Extract (AMPNE).

Implant insertion can evoke excessive inflammation which disrupts the healing process and potentially leads to complications such as implant rejection. Neutrophils and macrophages play a vital role in the early inflammatory phase of tissue repair, necessitating the study of cellular responses in host-implant interactions. In order to deepen the knowledge about these interactions, the response of neutrophils and macrophages to contact with selected biomaterials was examined in vitro on the basis of secretory response as well as reactive oxygen species/reactive nitrogen species (ROS/RNS) generation. Porcine neutrophils exposed to hydroxyapatite (HA) released more enzymes and generated higher levels of ROS/RNS compared to the control group. The addition of AMPNE diminished these responses. Although the results from porcine cells can provide valuable preliminary data, further validation using human cells or clinical studies would be necessary to fully extrapolate the findings to human medicine. Our study revealed that human neutrophils after contact of with HA increased the production of nitric oxide (NO) (10.00 ± 0.08 vs. control group 3.0 ± 0.11 µM, p < 0.05), while HAP or FAP did not elicit a significant response. Human macrophages cultured with HA produced more superoxide and NO, while HAP or FAP had a minimal effect, and curdlan reduced ROS/RNS generation. The addition of AMPNE to cultures with all biomaterials, except curdlan, reduced neutrophil activity, regardless of the peptides' origin. These results highlight the potential of antimicrobial peptides in modulating excessive biomaterial/host cell reactions involving neutrophils and macrophages, enhancing our understanding of immune reactions, and suggesting that AMPNE could regulate leukocyte response during implantation.

Intralipid blocks the entry of the SARS-Cov2/COVID/19 virus into cells by maintaining receptor a leucine-rich repeat containing 15 (LRRC15), an angiotensin-converting enzyme 2 receptor competitor.

The effectiveness of Intralipid against SARS-Cov2/COVID/19 with Multiple Organ Dysfunction Syndrome (MODS) prevention or regression is described in the original scientific paper [1]. Intralipid at Oxidative and Nitro-Galogenic stress in patients with SARS-Cov2 / COVID /19, favors the predominance of the membrane-cytoprotective action of Reactive Oxygen Species (ROS) / Reactive Nitrogen Species (ROS/RNS) over the membrane-to-destructive action, restoring the balance between [ROS /AS] / [RNS / ANOS]. Membrane-cytoprotective mechanism Intralipid is due to a decrease in ROS and RNS and an increase in the activity of the Antioxidant System (AS) and Anti Nitro Oxidant System (ANOS), stopping lipid peroxidation (LPO), reducing Electro-Ion Membrane Distress Syndrome (Maria&amp;Irina Vasilieva syndrome) [2], accelerates the regeneration of endothelial and epithelial cells of the alveolar acinus, restoring gas-respiratory metabolism and the predominance of physiological cell apoptosis over necrosis. Intralipid at SARS-Cov 2 / COVID / 19 opposes Microcirculatory Mitochondrial Distress syndrome (MMDS) by Microcirculatory - Mitochondrial Recruitment; as a result of which pCO2 (AVgap) &lt;6 mm Hg, since LPO decreases and at the level of mitochondrial membranes, improving the function of Mitochondrial permeability transition pore-dependent Ca uniporter, mPT pore, support energy metabolism, eliminating energy deficits, restoring Extreme / Abnormal myelopoiesis and impaired autophagy (mitophagy). Thus, Intralipid has been shown in the strategy of targeted treatment of LPO in Oxidative and Nitro-Galogenic stress in patients with SARS-Cov2 /COVID / 19.

Intralipid blocks the entry of the SARS-Cov2/COVID/19 virus into cells by maintaining receptor a leucine-rich repeat containing 15 (LRRC15), an angiotensin-converting enzyme 2 receptor competitor.

The effectiveness of Intralipid against SARS-Cov2/COVID/19 with Multiple Organ Dysfunction Syndrome (MODS) prevention or regression is described in the original scientific paper [1]. Intralipid at Oxidative and Nitro-Galogenic stress in patients with SARS-Cov2 / COVID /19, favors the predominance of the membrane-cytoprotective action of Reactive Oxygen Species (ROS) / Reactive Nitrogen Species (ROS/RNS) over the membrane-to-destructive action, restoring the balance between [ROS /AS] / [RNS / ANOS]. Membrane-cytoprotective mechanism Intralipid is due to a decrease in ROS and RNS and an increase in the activity of the Antioxidant System (AS) and Anti Nitro Oxidant System (ANOS), stopping lipid peroxidation (LPO), reducing Electro-Ion Membrane Distress Syndrome (Maria&amp;Irina Vasilieva syndrome) [2], accelerates the regeneration of endothelial and epithelial cells of the alveolar acinus, restoring gas-respiratory metabolism and the predominance of physiological cell apoptosis over necrosis. Intralipid at SARS-Cov 2 / COVID / 19 opposes Microcirculatory Mitochondrial Distress syndrome (MMDS) by Microcirculatory - Mitochondrial Recruitment; as a result of which pCO2 (AVgap) &lt;6 mm Hg, since LPO decreases and at the level of mitochondrial membranes, improving the function of Mitochondrial permeability transition pore-dependent Ca uniporter, mPT pore, support energy metabolism, eliminating energy deficits, restoring Extreme / Abnormal myelopoiesis and impaired autophagy (mitophagy). Thus, Intralipid has been shown in the strategy of targeted treatment of LPO in Oxidative and Nitro-Galogenic stress in patients with SARS-Cov2 /COVID / 19.

MicroRNAs, Long Non-Coding RNAs, and Circular RNAs in the Redox Control of Cell Senescence.

Cell senescence is critical in diverse aspects of organism life. It is involved in tissue development and homeostasis, as well as in tumor suppression. Consequently, it is tightly integrated with basic physiological processes during life. On the other hand, senescence is gradually being considered as a major contributor of organismal aging and age-related diseases. Increased oxidative stress is one of the main risk factors for cellular damages, and thus a driver of senescence. In fact, there is an intimate link between cell senescence and response to different types of cellular stress. Oxidative stress occurs when the production of reactive oxygen species/reactive nitrogen species (ROS/RNS) is not adequately detoxified by the antioxidant defense systems. Non-coding RNAs are endogenous transcripts that govern gene regulatory networks, thus impacting both physiological and pathological events. Among these molecules, microRNAs, long non-coding RNAs, and more recently circular RNAs are considered crucial mediators of almost all cellular processes, including those implicated in oxidative stress responses. Here, we will describe recent data on the link between ROS/RNS-induced senescence and the current knowledge on the role of non-coding RNAs in the senescence program.

MOTILIPERM Ameliorates Immobilization Stress-Induced Testicular Dysfunction via Inhibition of Oxidative Stress and Modulation of the Nrf2/HO-1 Pathway in SD Rats.

It is well established that physiological stress has an adverse effect on the male reproductive system. Experimental studies have demonstrated the promising effects of MOTILIPERM in male infertility. MOTILIPERM extract is composed of three crude medicinal herbs: Morinda officinalis How (Rubiaceae) roots, Allium cepa L. (Liliaceae) outer scales, and Cuscuta chinensis Lamark (convolvulaceae) seeds. The present study aimed to investigate the possible mechanisms responsible for the effects of MOTILIPERM on testicular dysfunction induced by immobilization stress. Fifty male Sprague Dawley rats were divided into five groups (10 rats each): a normal control group (CTR), a control group administered MOTILIPERM 200 mg/kg (M 200), an immobilization-induced stress control group (S), an immobilization-induced stress group administered MOTILIPERM 100 mg/kg (S + M 100), and MOTILIPERM 200 mg/kg (S + M 200). Stressed rats (n = 30) were subjected to stress by immobilization for 6 h by placing them in a Perspex restraint cage, while controls (n = 20) were maintained without disturbance. Rats were administrated 100 or 200 mg/kg MOTILIPERM once daily for 30 days 1 h prior to immobilization. At the end of the treatment period, we measured body and reproductive organ weight; sperm parameters; histopathological damage; reproductive hormone levels; steroidogenic acute regulatory protein (StAR); biomarkers of oxidative stress; and apoptosis markers. MOTILIPERM treatment improved testicular dysfunction by up-regulating (p < 0.05) sperm count, sperm motility, serum testosterone level, StAR protein level, Johnsen score, and spermatogenic cell density in stressed rats. MOTILIPERM decreased oxidative stress by increasing (p < 0.05) testicular superoxide dismutase (SOD), glutathione peroxidase (GPx), glutathione peroxidase-4 (GPx 4), catalase, nuclear factor erythroid 2-related factor 2 (Nrf2), and heme oxygenase 1 (HO-1) levels and decreasing (p < 0.05) malondialdehyde (MDA) and reactive oxygen species/reactive nitrogen species (ROS/RNS) levels. Furthermore, MOTILIPERM down-regulated (p < 0.05) cleaved caspase 3 and BCL2 associated X protein (Bax) levels; increased pro caspase-3 and B-cell lymphoma 2 (Bcl-2) levels; and upregulated testicular germ cell proliferation in stressed rats. The number of terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL)-positive cells and serum luteinizing hormone (LH) and follicle stimulating hormone (FSH) levels also significantly (p < 0.05) decreased after pretreatment with MOTILIPERM in stressed rats. Collectively, our results suggest that, in immobilization-mediated stress-induced testicular dysfunction, MOTILIPERM sustains normal spermatogenesis via antioxidant and anti-apoptotic activities by activating the NRF/HO-1 signaling pathway.

Role of ROS/RNS in Preeclampsia: Are Connexins the Missing Piece?

Preeclampsia is a pregnancy complication that appears after 20 weeks of gestation and is characterized by hypertension and proteinuria, affecting both mother and offspring. The cellular and molecular mechanisms that cause the development of preeclampsia are poorly understood. An important feature of preeclampsia is an increase in oxygen and nitrogen derived free radicals (reactive oxygen species/reactive nitrogen species (ROS/RNS), which seem to be central players setting the development and progression of preeclampsia. Cell-to-cell communication may be disrupted as well. Connexins (Cxs), a family of transmembrane proteins that form hemichannels and gap junction channels (GJCs), are essential in paracrine and autocrine cell communication, allowing the movement of signaling molecules between cells as well as between the cytoplasm and the extracellular media. GJCs and hemichannels are fundamental for communication between endothelial and smooth muscle cells and, therefore, in the control of vascular contraction and relaxation. In systemic vasculature, the activity of GJCs and hemichannels is modulated by ROS and RNS. Cxs participate in the development of the placenta and are expressed in placental vasculature. However, it is unknown whether Cxs are modulated by ROS/RNS in the placenta, or whether this potential modulation contributes to the pathogenesis of preeclampsia. Our review addresses the possible role of Cxs in preeclampsia, and the plausible modulation of Cxs-formed channels by ROS and RNS. We suggest these factors may contribute to the development of preeclampsia.

Interferon Gamma Reprograms Host Mitochondrial Metabolism through Inhibition of Complex II To Control Intracellular Bacterial Replication.

The mechanisms by which interferon gamma (IFN-γ) controls the replication of cytosolic pathogens independent of responses, such as the generation of reactive oxygen species/reactive nitrogen species (ROS/RNS), have not been fully elucidated. In the current study, we developed a model using Francisella tularensis, the causative agent of tularemia, in which pathways triggered by IFN-γ commonly associated with bacterial control were not required. Using this model, we demonstrated that IFN-γ-mediated production of itaconate and its ability to impair host mitochondrial function, independent of activity on the pathogen, were central for the restriction of bacterial replication in vitro and in vivo We then demonstrate that IFN-γ-driven itaconate production was dispensable, as directly targeting complex II using cell membrane-permeable metabolites also controlled infection. Together, these findings show that while reprogramming of mitochondrial metabolism is a key factor in IFN-γ control of intracellular bacteria, the development of antimicrobial strategies based on targeting host mitochondrial metabolism independent of this cytokine may be an effective therapeutic approach.

Numerical modelling of the effects of cold atmospheric plasma on mitochondrial redox homeostasis and energy metabolism

A biochemical reaction model clarifies for the first time how cold atmospheric plasmas (CAPs) affect mitochondrial redox homeostasis and energy metabolism. Fundamental mitochondrial functions in pyruvic acid oxidation, the tricarboxylic acid (TCA) cycle and oxidative phosphorylation involving the respiratory chain (RC), adenosine triphosphate/adenosine diphosphate (ATP/ADP) synthesis machinery and reactive oxygen species/reactive nitrogen species (ROS/RNS)-mediated mechanisms are numerically simulated. The effects of CAP irradiation are modelled as 1) the influx of hydrogen peroxide (H{}_{2}O{}_{2}) to an ROS regulation system and 2) the change in mitochondrial transmembrane potential induced by RNS on membrane permeability. The CAP-induced stress modifies the dynamics of intramitochondrial H{}_{2}O{}_{2} and superoxide anions, i.e., the rhythm and shape of ROS oscillation are disturbed by H{}_{2}O{}_{2} infusion. Furthermore, CAPs control the ROS oscillatory behaviour, nicotinamide adenine dinucleotide redox state and ATP/ADP conversion through the reaction mixture over the RC, the TCA cycle and ROS regulation system. CAPs even induce a homeostatic or irreversible state transition in cell metabolism. The present computational model demonstrates that CAPs crucially affect essential mitochondrial functions, which in turn affect redox signalling, metabolic cooporation and cell fate decision of survival or death.

Transcriptional Homeostasis of Oxidative Stress-Related Pathways in Altered Gravity.

Whereby several types of cultured cells are sensitive to gravity, the immune system belongs to the most affected systems during spaceflight. Since reactive oxygen species/reactive nitrogen species (ROS/RNS) are serving as signals of cellular homeostasis, particularly in the cells of the immune system, we investigated the immediate effect of altered gravity on the transcription of 86 genes involved in reactive oxygen species metabolism, antioxidative systems, and cellular response to oxidative stress, using parabolic flight and suborbital ballistic rocket experiments and microarray analysis. In human myelomonocytic U937 cells, we detected a rapid response of 19.8% of all of the investigated oxidative stress-related transcripts to 1.8 g of hypergravity and 1.1% to microgravity as early as after 20 s. Nearly all (97.2%) of the initially altered transcripts adapted after 75 s of hypergravity (max. 13.5 g), and 100% adapted after 5 min of microgravity. After the almost complete adaptation of initially altered transcripts, a significant second pool of differentially expressed transcripts appeared. In contrast, we detected nearly no response of oxidative stress-related transcripts in human Jurkat T cells to altered gravity. In conclusion, we assume a very well-regulated homeostasis and transcriptional stability of oxidative stress-related pathways in altered gravity in cells of the human immune system.

3-morpholinosydnonimine (SIN-1)-induced oxidative stress leads to necrosis in hypertrophic chondrocytes in vitro

Chondrocyte is targeted for disruption in Osteoarthritis (OA) and Kashin-Beck Disease (KBD), and chondrocyte death in cartilage may contribute to the progression of OA and KBD. Oxidative stress leads to increased risk for OA. Previous work in our laboratory implicates oxidative stress as a potential mediator in children with KBD. While these studies suggest a role for oxidative stress in the modulation of OA and KBD, the direct effects of reactive oxygen species/reactive nitrogen species (ROS/RNS) on the stability of this domain remain unclear. Here, we demonstrate that oxidative stress, as induced through treatment with 3-morpholinosydnonimine (SIN-1), a spontaneous ROS/RNS generator, decreased the cell viability in hypertrophic chondrocytes in a dose- and time- dependent manner. SIN-1 induced necrosis in hypertrophic chondrocytes, whereas triggered apoptosis in non-hypertrophic cells of non-differentiated ATDC5 cells and C28/I2 cells. Ultrastructural analysis of hypertrophic chondrocyte treated with SIN-1 revealed morphological changes, such as plasma membrane breakdown, generalized swelling of the cytoplasm and organelles, even to disappearance. Moreover, SIN-1 induced chondronecrosis in the deep zone of engineered cartilage tissue, such as cell-free vacancy and "red ghost" cells.Overall, we demonstrate for the first time that oxidative stress, as induced through exogenous ROS/RNS, leads to necrosis in hypertrophic chondrocytes. Oxidative stress-mediated necrotic cell death contributes to chondronecrosis in the deep zone of cartilage in both OA and KBD.

A Highly Selective and Sensitive Ratiometric Fluorescent Probe for Hypochlorite and Its Application

A novel water-soluble hybrid fluorescein-coumarin dye(FLNC) fluorescence ratiometric probe to recognize OCl− was designed and synthesized. The most prominent feature of the probe is that the molecular structure contains two different fluorophores, and the fluorescein moiety was connected to coumarin moiety by diacylhydrazine linker. When OCl− was added, the two fluorophores in the FLNC probe could have a rapid respond simultaneously, with high selectivity and sensitivity towards OCl– among other reactive oxygen species/reactive nitrogen species(ROS/RNS) and anions, and the detection limit was measured to be 0.30 μmol/L. In addition, the color changes of the solution could be observed by the naked eye and have a short response time(completed within 1 min), which suggestes that the probe FLNC has the potential to be used for real-time detection.

Para-Hydroxy Thiophenol-Coated CdSe/ZnS Quantum Dots as a Turn-On Fluorescent Probe for H2O2 Detection in Aqueous Media

Since hydrogen peroxide plays an important role in various fields, a facile, simple, highly selective, and stable analytic method for H2O2 is desirable. Semiconductor quantum dots (QDs) have acted as a potential alternative for organic fluorophores in fluorescence analytical fields due to their superior optical properties. Herein, we report hydrophilic p-hydroxy thiophenol (p-HTP) coated CdSe/ZnS QDs (denoted as p-HTP-QDs) acting as a selective fluorescence ‘turn-on’ probe for H2O2 in aqueous media. The obtained p-HTP-QD probe exhibits weak fluorescence, which stems from hole transfer from the QDs to p-HTP. The presence of H2O2 induces an oxidative structural transformation of p-HTP in p-HTP-QDs from a phenol structure to an α-hydroxy ketone derivative, which extremely reduces the driving force for hole transfer. Thus, the QDs photoluminescence (PL) was re-switched on. Under optimized conditions, an excellent linear relationship between fluorescence response and H2O2 concentration could be produced with a linear range from 0.309 to 4.900mM. The limit of detection of this probe was found to be 0.135mM. Moreover, the present probe exhibited a high selectivity of H2O2 over other reactive oxygen species/reactive nitrogen species (ROS/RNS) and was successfully used in the detection of H2O2 in real water samples.

Flow cytometic analysis of Penaeus monodon haemocyte responses to poly I:C

This study was aimed at investigating the cellular responses of Penaeus monodon haemocytes to poly I:C stimulation using flow cytometric assay. Total haemocyte count (THC), percentages of different haemocyte subpopulations [hyaline cells (HC), semigranular cells (SGC) and granular cells (GC)], non-specific esterase activity (EA), total reactive oxygen species/reactive nitrogen species (ROS/RNS) production, nitric oxide (NO) production, apoptotic haemocyte ratio and plasmic phenoloxidase (PO) activity were determined in poly I:C-injected shrimp. Results showed that poly I:C at a low dose (5 μg shrimp−1) caused obvious increases in THC, GC proportion, ROS/RNS production and NO production, but had no significant effect on EA, apoptosis and PO activity. In the early stage of poly I:C injection at a high dose (20 μg shrimp−1), THC and GC proportion improvements could also be observed, suggesting that GC might be induced to release from hemocytopoietic or other tissues to participate in immune response, and this subpopulation might be the main cell type involved in the cellular defence against virus. In the later period, proportions of both GC and SGC reduced paralleled by THC reduction, indicating that depletion of GC and SGC was mainly contributed to the reduced count of circulating haemocyte. Obvious increases in ROS/RNS production and NO production were induced in haemocyte of shrimp under a high dose of poly I:C stimulation, but only slight rise of EA and suppression of PO activity could be observed in poly I:C-stimulated shrimp, suggesting that ROS/RNS-dependent system was vital in the immune defence of shrimp against virus. On the other hand, increase of apoptotic haemocyte ratio and THC reduction were presented after the drastic increases of ROS/RNS and NO productions, implying that the stimulated ROS/RNS might be excess and harmful, and was the major factor for the haemocyte apoptosis and depletion. THC recovered after 48 h injection, while haemocyte apoptosis also returned to the control level, suggesting that apoptosis might be contributed to eliminate damaged, weak or infected haemocytes to renew the circulating haemocytes, and it could be considered as an important defending strategy against virus.

Oxidative burst activity in haemocytes of the freshwater prawn Macrobrachium rosenbergii

Oxidative burst, release of reactive oxygen species/reactive nitrogen species (ROS/RNS) contributed to microorganisms killing, is a vital immune response of crustacean haemocyte. Three morphologic haemocyte types (hyaline cells, HC; semigranular cells, SGC; granular cells, GC) have been defined in crustaceans, and found to play different roles in immune defense. However, oxidative burst activities of different haemocyte subpopulations in crustaceans are currently not documented. In the present study, we investigated the oxidative burst activities of the three haemocyte types in the freshwater prawn Macrobrachium rosenbergii using the common ROS fluorescent probe dichlorofluorescin-diacetate (DCFH-DA). Nitric oxide (NO) donor sodium nitroprusside (SNP) improved the DCF fluorescence in haemocytes, while NO scavenger C-PTIO (2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide) and NO-synthase inhibitor NG-monomethyl-l-arginine (L-NMMA) reduced the fluorescence, suggesting that DCF fluorescence intensity could also be modified by intracellular NO level and activity of NO-synthase pathway. ROS/RNS was also produced in the untreated haemocytes. GC contained most non-induced ROS/RNS production, while oxidative activity of HC was rather weak. No significant impact of PMA could be observed on ROS/RNS level in all the three cell types. Both zymosan A (ZA) and lipopolysaccharide (LPS) significantly triggered the production of ROS/RNS in SGC and GC, whereas they had no effect on those of HC, suggesting that SGC and GC were the primary cell types involved in pathogens killing by ROS/RNS pathway. Cytochalasin B (Cyt B) inhibited the ZA-induced ROS/RNS production, but could not change the ROS/RNS level stimulated by LPS. For unstimulated haemocytes, ROS/RNS productions decreased 29.6%, 44.1% and 48.6% in SGC, and decreased 44.5%, 28.4% and 57.3% in GC, in the presence of L-NMMA, Fccp and DPI respectively, whereas apocynin could not modulate DCF fluorescence in both SGC and GC, suggesting that mitochondrial oxidative pathway was relatively more dominant in SGC, and NO-synthase (NOS) pathway appeared more active in GC. For LPS-stimulated haemocytes, oxidative activities decreased 22.9%, 42.9%, 29.6% and 60.0% in SGC, and reduced 40.6%, 25.2%, 26.7% and 70.6% in GC with the presence of L-NMMA, apocynin, Fccp and DPI respectively, suggesting that NADPH-oxidase (NOX) pathway in both SGC and GC was activated by LPS, and it became the predominant oxidative pathway in stimulated SGC, while NOS pathway was the relative main source for ROS/RNS production in stimulated GC.