IS A DECREASED NITRIC OXIDE CONCENTRATION AFTER TRIAMCINOLONE ACETONIDE INTRAVITREAL INJECTION ONE OF THE REASONS FOR INTRAOCULAR PRESSURE RISE?

Diabetic macular edema is the most common cause of vision loss in patients affected by diabetes mellitus. For eyes with persistent retinal thickening despite anti-VEGF therapy, treatment with intravitreal triamcinolone may be considered, especially in pseudophakic eyes. The aim of this study was to examine aqueous humor nitric oxide concentration changes in pseudophakic eyes with persistent diffuse diabetic macular edema after intravitreal injection of triamcinolone acetonide, as well as the potential impact of these changes on the intraocular pressure values. In 10 pseudophakic eyes with persistent diffuse diabetic macular edema, paracentesis of anterior chamber with aspiration of aqueous humor and nitric oxide concentration measurements were done on the day of the intravitreal application of 20 mg triamcinolone acetonide, and after 1, 3, 6 and 9 months. Also, we were recording intraocular pressure values before the intravitreal triamcinolone acetonide injection and during the next 9 months. One month after the intravitreal triamcinolone acetonide injection, we noticed a decrease of nitric oxide concentration (45.37±5.55 µmol/L) by 31.79% compared to the initial values (66.52±7.66 µmol/L). After that, nitric oxide concentrations began to rise slightly, and at the end of the ninth month the mean nitric oxide concentration was similar to that recorded at the beginning of the study. Intraocular pressure values had increasing trend one month after the intravitreal triamcinolone acetonide injection (23.70±4.08 mm Hg) compared to the initial values (16.21±1.55 mm Hg), but after nine months these values returned to normal levels. Decreased concentration of nitric oxide could be one of the reasons for increased intraocular pressure after intravitreal application of triamcinolone acetonide in the treatment of diffuse diabetic macular edema.

QUALITY OF LIFE IN DYSPHONIC CHILDREN MEASURED ON PEDIATRIC VOICE-RELATED QUALITY OF LIFE (PVRQOL) SCALE IN SERBIA.

Hoarseness occurs in children of both genders, from the earliest age and beyond, and is caused by improper use or overuse of vocal apparatus. The study included 91 hoarse children aged 6-12 (study group) and 243 healthy children (control group) of the same age. The study group underwent detailed medical history, phoniatric examination, larynx fiber endoscopy, allergy treatment, audiologic treatment, and pulmonary treatment. Pediatric Voice-Related Quality of Life questionnaire, Serbian version (PVRQOL) was completed by parents of both groups of children. We did not find statistically significant differences in the hoarse children based on diagnosis (muscle tension disorder and vocal fold nodules) and age in physical domain, socio-emotional and global domain score (p>0.01). The results showed that parents did not recognize hoarseness as a health problem in children. There were significant gender differences in the group of children with hoarseness, i.e. parents in all three PVRQOL questionnaire domains recognized hoarseness as a significant health problem in girls, but not in boys. The presence of hoarseness impairs the quality of life in pediatric population. Social and emotional domains indicated greater impact in boys.

THE EFFECT OF PRIMARY ARGON LASER TRABECULOPLASTY ON INTRAOCULAR PRESSURE REDUCTION AND QUALITY OF LIFE IN PATIENTS WITH PSEUDOEXFOLIATION GLAUCOMA.

The aim of the study was to assess the impact of primary argon laser trabeculoplasty (ALT) on intraocular pressure (IOP) lowering and quality of life improvement in patients with pseudoexfoliative glaucoma. Sixty patients with newly diagnosed pseudoexfoliative glaucoma who underwent primary ALT (group 2) or medication therapy (group 1) were followed-up. The effect of ALT on IOP reduction, dry eye development and number of antiglaucoma drugs used was examined. Patients were examined at the beginning of the study and then after 6, 12, and 18 months. A statistically significant difference between IOP values was observed throughout the 18-month follow-up, with the highest significance recorded 6 months after ALT (p=0.009). Twelve months after the start of the study, the TBUT value was 6.0±0.8 s in group 1 and 8.4±0.7 s in group 2. In group 2, the value of Schirmer test was constantly above 10 millimeters. The number of antiglaucoma medications used in group 1 was statistically significantly higher as compared to group 2 throughout the 18-month study period. ALT was found to be better choice for temporary regulation of IOP in patients with pseudoexfoliative glaucoma.

Ganglion Cell-Inner Plexiform Layer Thickness in Different Glaucoma Stages Measured by Optical Coherence Tomography.

AIM: To compare ganglion cell (GCL) and inner plexiform layer (IPL) thickness in patients at different stages of primary open-angle glaucoma (POAG), determine their sensitivity and specificity values, and correlate thickness values with mean deviations (MD). METHODS: This prospective, cross- sectional study was conducted in a group of patients with confirmed POAG who were compared to an age- and gender-matched control group. Glaucomatous damage was classified according to the Hodapp-Parrish-Anderson scale: glaucoma stage 1 (early), glaucoma stage 2 (moderate), and glaucoma stage 3 (severe). The average, minimum, and all 6 sectoral (superotemporal, superior, superonasal, inferonasal, inferior, and inferotemporal) GCL + IPL thicknesses were measured and compared between groups. RESULTS: The average GCL + IPL thickness of 154 eyes of 93 patients in glaucoma stages 1, 2, 3, and 94 eyes of 47 persons in the control group were 76.79 ± 8.05, 65.90 ± 7.92, 57.38 ± 10.00, and 86.01 ± 3.68 µm, respectively. There were statistically significant differences in the average, minimum, and all 6 sectoral GCL + IPL values among the groups. The areas under the receiver operating characteristic curve for average and minimum GCL + IPL thickness values were 0.93 and 0.94, respectively, sensitivity 91.5 and 88.3%, and specificity 98.9 and 100%, respectively. Both thickness values showed significant correlations with MD. Each micrometer decrease in the average GCL + IPL thickness was associated with a 0.54-dB loss in MD. CONCLUSION: GCL + IPL layer thickness is a highly specific and sensitive parameter in differentiating glaucomatous from healthy eyes showing progressive damage as glaucoma worsens. Loss of this layer is highly correlated with overall loss of visual field sensitivity.

Quantitative Analysis of Uveitis Macular Edema in Multiple Sclerosis Patients Receiving Deep Posterior Sub-Tenon Triamcinolone Acetonide Injection.

AIM: We aimed to analyze the effects of adjunctive posterior sub-Tenon capsule triamcinolone acetonide injection in the treatment of intermediate uveitis macular edema in multiple sclerosis patients that could not be controlled by systemic corticosteroid medications and immunomodulators. METHODS: The study included 30 eyes of 25 patients with multiple sclerosis who received a posterior sub-Tenon injection of 40 mg/mL triamcinolone acetonide. Parameters monitored for therapy efficiency were best-corrected visual acuity, intraocular pressure, central foveal thickness (CFT), and fluorescein angiography (FA) scores. RESULTS: Mean best-corrected visual acuity was significantly improved at the control visit 0.15 ± 0.10 versus baseline 0.40 ± 0.20 logMAR (p < 0.05). Six eyes showed intraocular pressure spikes requiring topical antiglaucomatous treatment. Mean CFT and FA scores were significantly decreased versus baseline (CFT: 345 ± 50 µm; FA score: 3.4 ± 1.0) compared with the 12-week control visit (CFT: 219 ± 35 µm; FA score: 1.6 ± 1.1; p < 0.001). CONCLUSION: Posterior sub-Tenon injection of triamcinolone acetonide significantly improved visual acuity and decrease macular edema in patients with intermediate uveitis associated with multiple sclerosis.

A Rare Case of Infectious Multifocal Serpiginoid Choroiditis

Multifocal serpiginoid choroiditis is an infectious variety of serpiginous choroiditis. The disease is characterized by infectious etiology and overlapping clinical features in an intermediary form of acute posterior multifocal placoid pigment epitheliopathy and serpiginous choroiditis. In a 33-year-old patient, bilateral multiple placoid partially confluent chorioretinal lesions were diagnosed after a febrile flu-like episode. On the right eye, there was a progressive decrease in visual acuity. Later, the lesions had a prolonged progressive devastating clinical course and widespread distribution of placoid lesions, and took the form of serpiginoid choroiditis. We conducted extensive laboratory work-up and ancillary investigation for granulomatous diseases such as tuberculosis and sarcoidosis, and the results were not consistent with these entities. Systemic medical work-up revealed a history of exposure to the human immunodeficiency virus, herpes simplex virus 1, varicella zoster virus and cytomegalovirus. The titer of Mycoplasma pneumoniae IgM antibodies was positive. After serological analysis positive for Mycoplasma pneumoniae, systemic antibiotic therapy and anti-inflammatory doses of corticosteroids were administered. Improvement of visual acuity after the introduction of causal antibiotic therapy in combination with anti-inflammatory therapy confirmed our suspicion that Mycoplasma pneumoniae was the etiologic cause of multifocal serpiginoid choroiditis.

Blue-light-driven photoactivity of L-cysteine-modified graphene quantum dots and their antibacterial effects.

The widespread abuse of traditional antibiotics has led to a global rise in antibiotic-resistant bacteria, which give in return unprecedented health risks. Therefore, there is a large and urgent need for the development of new, smart antibacterial agents able to efficiently kill or inhibit bacterial growth. In this study, we investigated the antibacterial activity of S, N-doped Graphene Quantum Dots (GQDs) as a light-triggered antibacterial agent. Gamma irradiation was employed as a tool to achieve one-step modification of GQDs in the presence of L-cysteine amino acid as a source of heteroatoms. X-ray Photoelectron Spectroscopy (XPS), nuclear magnetic resonance (NMR), and zeta potential measurements provided the necessary data to clarify the structure of modified dots and verify the introduction of both S- and N-atoms in GQDs structure, but also severe changes in the aromatic, sp2 domains. Namely, γ-irradiation caused a bonding of S atoms in 1.14 at.% mainly as thiol groups, and N in 1.81 at.% as amino groups, but sp2 contribution in GQD structure was lowered from 63.00 to 4.86 at.%, as measured in dots irradiated at a dose of 200 kGy. Fluorescence quenching measurements showed that L-cysteine-modified dots are able to bind to human serum albumin. The antibacterial activity of GQDs combined with 1 and 6 h of blue light (470 nm) irradiation was tested against 8 bacterial strains. GQD-cys-25 sample provided the best results, with minimum inhibitory concentration (MIC) as low as 125 µg/mL against S. aureus, E. faecalis, and E. coli after only 1 h of blue light exposure.

The Influence of Reaction Conditions on the Properties of Graphene Oxide.

The present study focuses on correlations between three parameters: (1) graphite particle size, (2) the ratio of graphite to oxidizing agent (KMnO4), and (3) the ratio of graphite to acid (H2SO4 and H3PO4), with the reaction yield, structure, and properties of graphene oxide (GO). The correlations are a challenge, as these three parameters can hardly be separated from each other due to the variations in the viscosity of the system. The larger the graphite particles, the higher the viscosity of GO. Decreasing the ratio of graphite to KMnO4 from 1:4 to 1:6 generally leads to a higher degree of oxidation and a higher reaction yield. However, the differences are very small. Increasing the graphite-to-acid-volume ratio from 1 g/60 mL to 1 g/80 mL, except for the smallest particles, reduced the degree of oxidation and slightly reduced the reaction yield. However, the reaction yield mainly depends on the extent of purification of GO by water, not on the reaction conditions. The large differences in the thermal decomposition of GO are mainly due to the bulk particle size and less to other parameters.

Quality of Life of Patients with Oral Squamous Cell Carcinoma.

Background: Oral squamous cell carcinoma (OSCC) has numerous physical, psychosocial and financial implications, which significantly affect patients' quality of life. We aimed to determine the health-related quality of life (HRQoL) and identify quality of life (QoL) predictors in patients with OSCC. Methods: We included 64 consecutive patients aged 40 to 80 yr treated for OSCC from Jan to Dec 2021. Health-related QoL was evaluated using the 30-item Cancer Quality of Life Questionnaire (QLQ-C30) and the 35-item Head and Neck Cancer-Quality of Life Questionnaire (QLQ-H&N35). The demographic questionnaire and clinical parameters were also presented. Results: The functioning scale in the QLQ-C30 questionnaire with the lowest average score was Global health status. The mean QLQ-C30 summary score (80.92 ± 10.4) was higher than the Global health status score (50.5 ± 22.2). In the QLQ-H&N35 questionnaire, the symptoms with highest scores were weight loss, dry mouth, and social eating. Linear regression analysis demonstrated that Global health status score was associated with education level [ß-coefficient = 19.33 (95% CI: 10.7-24.9, P=0.004], alcohol consumption [ß-coefficient=10.04 (95% CI: 4.5-14.8), P=0.023] and invasive surgical procedure [ß-coefficient=22.75 (95% CI: 15.0-30.5), P=0.002]. The QLQ-C30 summary score was associated with living alone [ß-coefficient= -20.05 (95% CI: -29.91-(-10.21), P=0.018], smoking status [ß-coefficient=4.35 (95% CI: 1.8-6.91), P=0.043] and alcohol consumption [ß-coefficient =4.59 (95% CI: 1.99-7.19), P=0.037]. Conclusion: We found several significant predictors of worse perception of HRQoL among patients with OSCC, which may be useful for specific prevention and treatment in order to achieve better QoL.

Determination of Photothermal and EMI Shielding Efficiency of Graphene-Silver Nanoparticle Composites Prepared under Low-Dose Gamma Irradiation.

Silver nanoparticles (Ag NPs) have been produced by low-dose (1-20 kGy) gamma irradiation of silver nitrate in the presence of graphene-based material (graphene oxide or electrochemically exfoliated graphene). The large surface area of those graphene-based materials combined with the presence of oxygen-containing functional groups on the surface provided successful nucleation and growth of Ag nanoparticles, which resulted in a uniformly covered graphene surface. The obtained Ag nanoparticles were spherical with a predominant size distribution of 10-50 nm for graphene oxide and 10-100 nm for electrochemically exfoliated graphene. The photothermal efficiency measurement showed a temperature increase upon exposure to a 532 nm laser for all samples and the highest photothermal efficiency was measured for the graphene oxide/Ag NP sample prepared at 5 kGy. Electromagnetic interference (EMI) shielding efficiency measurements showed poor shielding for the composites prepared with graphene oxide. On the other hand, all composites prepared with electrochemically exfoliated graphene showed EMI shielding to some extent, and the best performance was measured for the samples prepared at 5 and 20 kGy doses.

Toxicity induced by cumene hydroperoxide in leech Retzius nerve cells: the protective role of glutathione.

In the present study, we studied the ability of glutathione (GSH) to detoxify exogenously applied cumene hydroperoxide (CHP). Exposure of leech Retzius nerve cells to CHP (1.5 mM) induced a marked prolongation of the spontaneous spike potential of these cells. Early after depolarization, and a cardiac-like action potential with a rapid depolarization followed by a sustained depolarization or plateau, which is terminated by a rapid repolarization were recorded. GSH (0.2 mM) significantly inhibited the effects of CHP on the duration of the action potential and suppressed CHP-induced spontaneous repetitive activity. Voltage-clamp recordings showed that CHP (1.5 mM) caused significant changes in the outward potassium currents. The fast and slow steady part of the potassium outward current was reduced by 46% and 39%, respectively. GSH applied in a concentration of 0.2 mM partially blocked the effect of CHP on the calcium-activated potassium currents. The fast and slow calcium-activated potassium currents were suppressed by about 20% and 15%, respectively. These results suggest that the neurotoxic effect of CHP on spontaneous spike electrogenesis and calcium-activated potassium currents of leech Retzius nerve cells was reduced in the presence of GSH.

Lights and Dots toward Therapy-Carbon-Based Quantum Dots as New Agents for Photodynamic Therapy.

The large number of deaths induced by carcinoma and infections indicates that the need for new, better, targeted therapy is higher than ever. Apart from classical treatments and medication, photodynamic therapy (PDT) is one of the possible approaches to cure these clinical conditions. This strategy offers several advantages, such as lower toxicity, selective treatment, faster recovery time, avoidance of systemic toxic effects, and others. Unfortunately, there is a small number of agents that are approved for usage in clinical PDT. Novel, efficient, biocompatible PDT agents are, thus, highly desired. One of the most promising candidates is represented by the broad family of carbon-based quantum dots, such as graphene quantum dots (GQDs), carbon quantum dots (CQDs), carbon nanodots (CNDs), and carbonized polymer dots (CPDs). In this review paper, these new smart nanomaterials are discussed as potential PDT agents, detailing their toxicity in the dark, and when they are exposed to light, as well as their effects on carcinoma and bacterial cells. The photoinduced effects of carbon-based quantum dots on bacteria and viruses are particularly interesting, since dots usually generate several highly toxic reactive oxygen species under blue light. These species are acting as bombs on pathogen cells, causing various devastating and toxic effects on those targets.

Antioxidative and Photo-Induced Effects of Different Types of N-Doped Graphene Quantum Dots.

Due to the increasing number of bacterial infections and the development of resistivity toward antibiotics, new materials and approaches for treatments must be urgently developed. The production of new materials should be ecologically friendly considering overall pollution with chemicals and economically acceptable and accessible to the wide population. Thus, the possibility of using biocompatible graphene quantum dots (GQDs) as an agent in photodynamic therapy was studied. First, dots were obtained using electrochemical cutting of graphite. In only one synthetic step using gamma irradiation, GQDs were doped with N atoms without any reagent. Obtained dots showed blue photoluminescence, with a diameter of 19-89 nm and optical band gap of 3.23-4.73 eV, featuring oxygen-containing, amino, and amide functional groups. Dots showed antioxidative activity; they quenched •OH at a concentration of 10 µg·mL-1, scavenged DPPH• radicals even at 5 µg·mL-1, and caused discoloration of KMnO4 at 30 µg·mL-1. Under light irradiation, dots were able to produce singlet oxygen, which remained stable for 10 min. Photoinduced effects by GQDs were studied on several bacterial strains (Listeria monocytogenes, Bacillus cereus, clinical strains of Streptococcus mutans, S. pyogenes, and S. sangunis, Pseudomonas aeruginosa, and one yeast strain Candida albicans) but antibacterial effects were not noticed.

Mental Health and Quality of Life among Dental Students during COVID-19 Pandemic: A Cross-Sectional Study.

Students are particularly vulnerable from the mental health aspect, which was especially recognized during the COVID-19 pandemic. This study aimed to reveal the impact of COVID-19 on quality of life (QoL) and mental health among dental students. The study was conducted on a sample of 797 students (207 male and 592 female) with an average age of 21.7 ± 2.4, from the School of Dental Medicine, University of Belgrade. The measurements used in the study were the Demographic and Academic Questionnaire, Questionnaire about exposure to COVID-19, COVID-19-Impact on QoL Questionnaire (COV19-QoL), Generalized Anxiety Disorder 7-item (GAD-7) scale, and Patient Health Questionnaire (PHQ-9). The mean total score for COV19-QoL was 2.9 ± 0.9, while the diagnostic criteria of GAD-7 and depression met 19.9% and 31.4% of students, respectively. There was a positive and strong correlation between QoL, anxiety, and depression. During COVID-19, predictors for lower perceptions of QoL were female gender and death of close relatives (p = 0.049, p = 0.005, respectively). At the same time, predictors for GAD were female gender, living in dormitories, and death of close relatives (p = 0.019, p = 0.011, p = 0.028, respectively), while for depression they were year of study, living with parents, and death of close relatives due to COVID-19 (p = 0.012, p = 0.008, p = 0.029, respectively). The study showed that students' QoL and mental health during the pandemic were at high risk.

Gamma-Ray-Induced Structural Transformation of GQDs towards the Improvement of Their Optical Properties, Monitoring of Selected Toxic Compounds, and Photo-Induced Effects on Bacterial Strains.

Structural modification of different carbon-based nanomaterials is often necessary to improve their morphology and optical properties, particularly the incorporation of N-atoms in graphene quantum dots (GQDs). Here, a clean, simple, one-step, and eco-friendly method for N-doping of GQDs using gamma irradiation is reported. GQDs were irradiated in the presence of the different ethylenediamine (EDA) amounts (1 g, 5 g, and 10 g) and the highest % of N was detected in the presence of 10 g. N-doped GQDs emitted strong, blue photoluminescence (PL). Photoluminescence quantum yield was increased from 1.45, as obtained for non-irradiated dots, to 7.24% for those irradiated in the presence of 1 g of EDA. Modified GQDs were investigated as a PL probe for the detection of insecticide Carbofuran (2,2-Dimethyl-2,3-dihydro-1-benzofuran-7-yl methylcarbamate) and herbicide Amitrole (3-amino-1,2,4-triazole). The limit of detection was 5.4 µmol L-1 for Carbofuran. For the first time, Amitrole was detected by GQDs in a turn-off/turn-on mechanism using Pd(II) ions as a quenching agent. First, Pd(II) ions were quenched (turn-off) PL of GQDs, while after Amitrole addition, PL was recovered linearly with Amitrole concentration (turn-on). LOD was 2.03 µmol L-1. These results suggest that modified GQDs can be used as an efficient new material for Carbofuran and Amitrole detection. Furthermore, the phototoxicity of dots was investigated on both Gram-positive and Gram-negative bacterial strains. When bacterial cells were exposed to different GQD concentrations and illuminated with light of 470 nm wavelength, the toxic effects were not observed.

Shedding Light on Graphene Quantum Dots: Key Synthetic Strategies, Characterization Tools, and Cutting-Edge Applications.

During the last 20 years, the scientific community has shown growing interest towards carbonaceous nanomaterials due to their appealing mechanical, thermal, and optical features, depending on the specific nanoforms. Among these, graphene quantum dots (GQDs) recently emerged as one of the most promising nanomaterials due to their outstanding electrical properties, chemical stability, and intense and tunable photoluminescence, as it is witnessed by a booming number of reported applications, ranging from the biological field to the photovoltaic market. To date, a plethora of synthetic protocols have been investigated to modulate the portfolio of features that GQDs possess and to facilitate the use of these materials for target applications. Considering the number of publications and the rapid evolution of this flourishing field of research, this review aims at providing a broad overview of the most widely established synthetic protocols and offering a detailed review of some specific applications that are attracting researchers' interest.

Recent Advances in the Label-Free Characterization of Exosomes for Cancer Liquid Biopsy: From Scattering and Spectroscopy to Nanoindentation and Nanodevices.

Exosomes (EXOs) are nano-sized vesicles secreted by most cell types. They are abundant in bio-fluids and harbor specific molecular constituents from their parental cells. Due to these characteristics, EXOs have a great potential in cancer diagnostics for liquid biopsy and personalized medicine. Despite this unique potential, EXOs are not yet widely applied in clinical settings, with two main factors hindering their translational process in diagnostics. Firstly, conventional extraction methods are time-consuming, require large sample volumes and expensive equipment, and often do not provide high-purity samples. Secondly, characterization methods have some limitations, because they are often qualitative, need extensive labeling or complex sampling procedures that can induce artifacts. In this context, novel label-free approaches are rapidly emerging, and are holding potential to revolutionize EXO diagnostics. These methods include the use of nanodevices for EXO purification, and vibrational spectroscopies, scattering, and nanoindentation for characterization. In this progress report, we summarize recent key advances in label-free techniques for EXO purification and characterization. We point out that these methods contribute to reducing costs and processing times, provide complementary information compared to the conventional characterization techniques, and enhance flexibility, thus favoring the discovery of novel and unexplored EXO-based biomarkers. In this process, the impact of nanotechnology is systematically highlighted, showing how the effectiveness of these techniques can be enhanced using nanomaterials, such as plasmonic nanoparticles and nanostructured surfaces, which enable the exploitation of advanced physical phenomena occurring at the nanoscale level.

Facile Synthesis of L-Cysteine Functionalized Graphene Quantum Dots as a Bioimaging and Photosensitive Agent.

Nowadays, a larger number of aggressive and corrosive chemical reagents as well as toxic solvents are used to achieve structural modification and cleaning of the final products. These lead to the production of residual, waste chemicals, which are often reactive, cancerogenic, and toxic to the environment. This study shows a new approach to the modification of graphene quantum dots (GQDs) using gamma irradiation where the usage of reagents was avoided. We achieved the incorporation of S and N atoms in the GQD structure by selecting an aqueous solution of L-cysteine as an irradiation medium. GQDs were exposed to gamma-irradiation at doses of 25, 50 and 200 kGy. After irradiation, the optical, structural, and morphological properties, as well as the possibility of their use as an agent in bioimaging and photodynamic therapy, were studied. We measured an enhanced quantum yield of photoluminescence with the highest dose of 25 kGy (21.60%). Both S- and N-functional groups were detected in all gamma-irradiated GQDs: amino, amide, thiol, and thione. Spin trap electron paramagnetic resonance showed that GQDs irradiated with 25 kGy can generate singlet oxygen upon illumination. Bioimaging on HeLa cells showed the best visibility for cells treated with GQDs irradiated with 25 kGy, while cytotoxicity was not detected after treatment of HeLa cells with gamma-irradiated GQDs.

Graphene quantum dot antioxidant and proautophagic actions protect SH-SY5Y neuroblastoma cells from oxidative stress-mediated apoptotic death.

We investigated the ability of graphene quantum dot (GQD) nanoparticles to protect SH-SY5Y human neuroblastoma cells from oxidative/nitrosative stress induced by iron-nitrosyl complex sodium nitroprusside (SNP). GQD reduced SNP cytotoxicity by preventing mitochondrial depolarization, caspase-2 activation, and subsequent apoptotic death. Although GQD diminished the levels of nitric oxide (NO) in SNP-exposed cells, NO scavengers displayed only a slight protective effect, suggesting that NO quenching was not the main protective mechanism of GQD. GQD also reduced SNP-triggered increase in the intracellular levels of hydroxyl radical (•OH), superoxide anion (O2•-), and lipid peroxidation. Nonselective antioxidants, •OH scavenging, and iron chelators, but not superoxide dismutase, mimicked GQD cytoprotective activity, indicating that GQD protect cells by neutralizing •OH generated in the presence of SNP-released iron. Cellular internalization of GQD was required for optimal protection, since a removal of extracellular GQD by extensive washing only partly diminished their protective effect. Moreover, GQD cooperated with SNP to induce autophagy, as confirmed by the inhibition of autophagy-limiting Akt/PRAS40/mTOR signaling and increase in autophagy gene transcription, protein levels of proautophagic beclin-1 and LC3-II, formation of autophagic vesicles, and degradation of autophagic target p62. The antioxidant activity of GQD was not involved in autophagy induction, as antioxidants N-acetylcysteine and dimethyl sulfoxide failed to stimulate autophagy in SNP-exposed cells. Pharmacological inhibitors of early (wortmannin, 3-methyladenine) or late stages of autophagy (NH4Cl) efficiently reduced the protective effect of GQD. Therefore, the ability of GQD to prevent the in vitro neurotoxicity of SNP depends on both •OH/NO scavenging and induction of cytoprotective autophagy.

Oxidative stress-mediated hemolytic activity of solvent exchange-prepared fullerene (C60) nanoparticles.

The present study investigated the hemolytic properties of fullerene (C(60)) nanoparticles prepared by solvent exchange using tetrahydrofuran (nC(60)THF), or by mechanochemically assisted complexation with macrocyclic oligosaccharide gamma-cyclodextrin (nC(60)CDX) or the copolymer ethylene vinyl acetate-ethylene vinyl versatate (nC(60)EVA-EVV). The spectrophotometrical analysis of hemoglobin release revealed that only nC(60)THF, but not nC(60)CDX or nC(60)EVA-EVV, was able to cause lysis of human erythrocytes in a dose- and time-dependent manner. Atomic force microscopy revealed that nC(60)THF-mediated hemolysis was preceded by erythrocyte shrinkage and increase in cell surface roughness. A flow cytometric analysis confirmed a decrease in erythrocyte size and demonstrated a significant increase in reactive oxygen species production in red blood cells exposed to nC(60)THF. The nC(60)THF-triggered hemolytic activity was efficiently reduced by the antioxidants N-acetylcysteine and butylated hydroxyanisole, as well as by serum albumin, the most abundant protein in human blood plasma. These data indicate that nC(60)THF can cause serum albumin-preventable hemolysis through oxidative stress-mediated damage of the erythrocyte membrane.