Yes / Non-steroidal anti-inflammatory drugs (NSAIDs) inhibit COX enzyme activity which affects the inflammatory response. Inflammation is associated with increasing cancer incidence. Pre-clinical and clinical studies have shown that NSAID treatment could cause an anti-tumor effect in cancers. In the present study, blood was taken from healthy individuals (n = 17) and patients with respiratory diseases or lung cancer (n = 36). White blood cells (WBC) were treated with either a micro-suspension, i.e., bulk (B) or nano-suspension (N) of aspirin (ASP) or ibuprofen (IBU) up to 500 μg/ml in the comet assay and up to 125 μg/ml in the micronucleus assay. In this study results were compared against untreated lymphocytes and their corresponding treated groups. The results showed, that NSAIDs in their nano form significantly reduced the DNA damage in WBCs from lung cancer patients in bulk and nano compared to untreated lymphocytes. Also, there was a decrease in the level of DNA damage in the comet assay after treating WBCs from healthy individuals, asthma and COPD groups with aspirin N (ASP N) but not with IBU N. In addition, the number of micronuclei decreased after treatment with NSAIDs in their nano form (ASP N and IBU N) in the healthy as well as in the lung cancer group. However, this was not the case for micronucleus frequency in asthma and COPD patients. These data show that lymphocytes from different groups respond differently to treatment with ASP and IBU as measured by comet assay and micronucleus assay, and that the size of the suspended particles of the drugs affects responses. / The present study was part funded by United Kingdom India Education Research Initiative (UKERI) SA 07-067.

Yes / We investigated the protective role of myricetin bulk and nanoforms, against reactive oxygen species (ROS)-induced oxidative stress caused by hydrogen peroxide and tertiary-butyl hydro peroxide in lymphocytes in vitro from healthy individuals and those from pre-cancerous patients suffering with monoclonal gammopathy of undetermined significance (MGUS). The change in intracellular reactive oxygen species was measured once cells were treated with myricetin bulk forms and nanoforms with and without either hydrogen peroxide or tertiary-butyl hydro peroxide co-supplementation. The direct and indirect antioxidant activity of myricetin was spectrofluometrically measured using the fluorescent dye 2',7'-dichlorofluorescin diacetate and using the Comet assay, respectively. Hydrogen peroxide (50 µM) and tertiary-butyl hydro peroxide (300 µM) induced a higher level of reactive oxygen species-related DNA damage and strand breaks. Addition of myricetin nanoform (20 µM) and bulk (10 µM) form could, however, significantly prevent hydrogen peroxide- and tertiary-butyl hydro peroxide-induced oxidative imbalances and the nanoform was more effective. Glutathione levels were also quantified using a non-fluorescent dye. Results suggest that myricetin treatment had no significant effect on the cellular antioxidant enzyme, glutathione. The current study also investigates the effect of myricetin on the induction of double-strand breaks by staining the gamma-H2AX foci immunocytochemically. It was observed that myricetin does not induce double-strand breaks at basal levels rather demonstrated a protective effect.

The head and neck cancers (HNC) are a group of cancers that begin in the squamous cells that line the mucosal surfaces of the head and neck. Therefore, they are commonly known as squamous cell carcinoma of the head and neck. Squamous cell carcinoma of the maxillary sinus (MC) is a rare type of HNC, and it is a very aggressive tumour. This cancer is typically diagnosed at a very advanced stage and most patients have a poor survival rate and prognosis. This study is based on the synthesis and applications of gold nanoparticles (AuNPs) conjugated with hesperidin (Hsp) as a targeted drug delivery system. AuNPs are ideal for loading different drugs and delivering them to targets sites due to their stability, small size, substantial surface area, non-cytotoxic and inert nature. Hsp is a naturally occurring substance with anti-inflammatory and antioxidant capabilities. The main aim of this research was to develop a highly efficient and safer method to deliver Hsp to the target sites. The Hsp with poor solubility and bioavailability render it only slightly absorbed, requiring a delivery system to reach its therapeutic target. This study focused on the effects of 15μg/ml Hsp loaded on gold nanoparticles (Hsp-AuNPs) on 20 healthy individuals’ lymphocytes as compared to 20 HNC patients’ lymphocytes using the alkaline Comet assay. While enzyme-based Comet repair was performed on 5 healthy individuals’ lymphocytes as compared to 5 HNC patients’ lymphocytes. The Hsp-AuNPs reduced the DNA damage in HNC patients’ lymphocytes compared to the healthy lymphocytes (***p<0.001). Furthermore, the 15μg/ml of Hsp-AuNPs significantly reduced the oxidative stress caused by H2O2 and appeared to be effective in both groups using the Comet and Comet repair assay. Results from Comet and Comet repair assay were consistent. The human squamous cells of maxillary sinus (MC) were also treated with 5μg/ml of Hsp-AuNPs. The alkaline Comet assay results showed that Hsp-AuNPs induced DNA damage in MC cells (***p<0.001). Therefore, Hsp-AuNPs demonstrated the most substantial genotoxic effects and confirmed a possible anticancer agent. The Hsp was also used to treat lymphocytes from healthy individuals as compared to HNC patients’ lymphocytes they reduced the DNA damage, but they were less effective as compared to Hsp-AuNPs. Published data shows that using the AuNPs as a drug carrier has a more potent therapeutic effect against different diseases including cancer. Also, this study investigated the gene protective and genotoxic impact of bulk Hsp in Maxillary sinus carcinoma cells. The data obtained indicated that Hsp-AuNPs might possibly be effective for the treatment of MC and demonstrated the ability of Hsp-AuNPs to increase the DNA damage more than the bulk form of Hsp (***p<0.001). The outcomes of this study are consistent with the viewpoint that the Hsp-AuNPs might have a substantial role in cancer treatment, including MC. The concentration of 5μg/ml Hsp-AuNPs was used to treat the MC cells in Western blotting, and real-time polymerase chain reaction (qPCR) was based on a preliminary test for the optimal dose. The data obtained indicated that Hsp-AuNPs might potentially be effective for the treatment of HNC and showed the ability of Hsp-AuNPs to reduce DNA damage more than the bulk form of Hsp. Hsp-AuNPs has also shown anti-cancer potential in the MC cells by up-regulating the expression of p53, p21, PPAR gamma, and Caspase 3, at mRNA and protein levels by up-regulating the p53, PPAR gamma, Caspase 3, and p21 to mediate apoptosis and DNA repair in MC cells. The findings of this study are consistent with the view that the Hsp-AuNPs could have a significant role in cancer treatment, including HNC and MC.