Annie Abraham

Bis(3,5-diiodo-2,4,6-trihydroxyphenyl)squaraine: a novel candidate in photodynamic therapy for skin cancer models in vivo.

Photodynamic therapy (PDT) is based on the light-induced activation of a photosensitizer generating highly reactive oxygen species that induce tissue destruction in malignant tissues. The present study was carried out to assess the photosensitizing potential of bis(3,5-diiodo-2,4,6-trihydroxyphenyl)squaraine in PDT trials in vivo. Male Swiss albino mice were divided into five groups. Skin tumor was induced using 7,12-dimethylbenz(a)anthracene - DMBA in the animals of Groups II, III, IV and V, while animals of Group I served as the control. At the completion of 20 weeks of induction, the tumor bearing mice from Group III, IV and V were given an intraperitoneal injection with the squaraine dye (12.5mg/kg body weight). After 24h, in the Group IV and V animals, the tumor area was exposed to visible light from a 1000W halogen lamp. The mice from groups I to IV were sacrificed two weeks after the PDT treatment and the marker enzymes (myeloperoxidase [MPO], beta-d-glucuronidase, rhodanese, lactate dehydrogenase [LDH], hexokinase, sialic acid and caspase) were assayed in tumor and normal tissues. Animals from Group V were sacrificed after 90 days of PDT treatment and the above parameters were recorded. Reduction in tumor volume and reversal of biochemical markers to near normal levels were observed in the treatment groups. The study assumes importance as it is the first report on PDT-a novel modality, using a squaraine dye for skin cancer therapy in vivo. The uniqueness of the mode of treatment lies in the selective uptake of squaraine dye by the cancer cells and their selective destruction using PDT without affecting the neighbouring normal cells, which is much advantageous over radiation therapy now frequently used. Also in skin cancer models, the progression/cure can be visualized by the naked eye which is another point of advantage, while seeking new modalities for the treatment of cancer.

Vitex negundo attenuates calpain activation and cataractogenesis in selenite models.

Recent investigations have shown that phytochemical antioxidants can scavenge free radicals and prevent various diseases. Cataract is the leading cause of blindness and is associated with oxidative damage of the lens. Selenite-induced cataract in rat pups is an excellent mimic of oxidative stress-induced cataract. Selenite cataract is associated with oxidative stress, loss of calcium homeostasis, calpain activation and protein insolubilization in the lens. Our present study focuses on the isolation of flavonoids from Vitex negundo and to assess its efficacy in preventing these changes in the lens of selenite-induced cataract models. Eight-day-old Sprague-Dawley rat pups were used for the study and divided into four groups: Control (G I), Sodium selenite-induced (G II), Sodium selenite+quercetin treated (G III), Sodium selenite+flavonoids from Vitex negundo (FVN) (G IV). Cataract was induced by a single subcutaneous injection of Sodium selenite (4 mg/Kg body weight) on the 10th day. Treatment groups received quercetin (1.0mg/Kg body weight) and FVN (1.0mg/Kg body weight) intraperitoneally from 8th to 15th day. Cataract was visualized from the 16th day. Morphological examination of the rat lenses revealed no opacification in G I and mild opacification in G III and G IV (stage 1) whereas dense opacification in G II (stage 4-6). The activities of superoxide dismutase (SOD), catalase, Ca(2+)ATPase, concentration of reduced glutathione (GSH) and protein sulfhydryl content were significantly increased in G III and G IV compared to G II, while decreased activities of calpains, lower concentration of calcium and thiobarbituric acid reactive substances (TBARS) were observed in G III and IV as compared to G II. Lens protein profile of water soluble proteins showed normal levels of expression in treated groups compared to that of selenite-induced rats. These results indicate good antioxidant and therapeutic potential of FVN in modulating biochemical parameters against selenite-induced cataract, which have been reported in this paper for the first time.

Biochemical response in rats to the addition of curry leaf (Murraya koenigii) and mustard seeds (Brassica juncea) to the diet

Three groups with 12 weanling male albino rats were group fed for 90 days on a standard laboratory rat diet plus 20% coconut oil either without a supplement or with the addition of 10% curry leaf or 10% mustard seeds. Feed was offered at a level of 10% body weight. At the end of the trial, measurements were made on the total serum cholesterol, high density lipoproteins, low density lipoproteins, and very low density lipoproteins fractions, release of lipoproteins into the circulation, lecithin cholesterol acyl transferase activity and lipoprotein lipase activity. Feed intake and mean body weight of the rats on each treatment was not significantly different. Both spices resulted in a reduction in total serum cholesterol and LDL+VLDL, an increase in the HDL, lower release of lipoproteins into the circulation and an increase in the LCAT activity.

Laser immunotherapy with gold nanorods causes selective killing of tumour cells

Therapeutic approaches that exploit nanoparticles to deliver drugs selectively to cancer cells are currently considered one of the most promising avenues in the area of cancer therapeutics. Recently, gold nanorods (AuNRs) have shown promising biological applications due to their unique electronic and optical properties. In this paper, we have demonstrated the anti-cancer potential of gold nanorods with low power laser light. Gold nanorods (AuNRs), surface modified with poly (styrene sulfonate) PSS and functionalized with epidermal growth factor receptor antibody conjugated with gold nanorods (anti-EGFR-AuNRs) were successfully synthesised and characterized by UV-Visible-NIR spectrophotometry and High Resolution Transmission Electron Microscopy (HR-TEM). Inductively Coupled Plasmon Atomic Emission Spectrometry (ICP-AES) and Immunofluorescence studies confirmed the efficient uptake of these functionalized gold nanorods by human squamous carcinoma cells, A431. The in vitro photothermal therapy was conducted in four groups - control, laser alone, unconjugated AuNRs with laser and anti-EGFR conjugated AuNRs with laser. Phase contrast images have revealed cell morphology changes and cell death after the laser irradiation. In order to determine whether the cell death occur due to apoptosis or necrosis, we have evaluated the biochemical parameters such as lactate dehydrogenase release, reactive oxygen species level, mitochondrial membrane potential and caspase-3 activity. Flow cytometry analysis have shown the cell cycle changes after laser irradiation with antibody conjugated gold nanorods. Thus the results of our experiments confirmed that immunolabeled gold nanorods can selectively destruct the cancer cells and induce its apoptosis through ROS mediated mitochondrial pathway under low power laser exposure.

Moringa oleifera Prevents Selenite-Induced Cataractogenesis in Rat Pups

PURPOSE The aim of this study was to investigate the protective effects of the flavonoid fraction of Moringa oleifera leaves (FMO) on selenite cataract in vivo. METHODS Rat pups of Sprague-Dawley strain initially weighing 10–12 g on day 8 were used for the study and grouped as control (G I), selenite induced (G II), and FMO treated (G III). The rat pups in G II and G III received a single subcutaneous injection of sodium selenite (4 µg/g body weight) on day 10 and G III was administered with FMO (2.5 µg/g body weight) from day 8 to 15. Cataract was visualized from day 16. The development of cataract was assessed and rat lenses were analyzed for the activities of antioxidant enzymes (superoxide dismutase and catalase), generation of reactive oxygen species, reduced glutathione, protein oxidation, and lipid peroxidation. FMO was subjected to in vitro antioxidant assays (2,2-diphenyl-picrylhydrazyl and superoxide scavenging assays). RESULTS The total phenolic content of FMO was 4.4 mg of catechin equivalent/g dried plant material. The extract showed remarkable activity on 2,2-diphenyl-picrylhydrazyl (IC50 36 µg/mL) and in superoxide radical (IC50 33.81µg/mL) scavenging assays. FMO effectively prevented the morphological changes and oxidative damage in lens. FMO maintained the activities of antioxidant enzymes and sulfhydryl content and prevented reactive oxygen species generation and lipid peroxidation. CONCLUSIONS FMO was effective in preventing cataractogenesis in selenite model by enhancing the activities of antioxidant enzyme, reducing the intensity of lipid peroxidation, and inhibiting free radical generation.

Murraya koenigii and Brassica juncea - alterations on lipid profile in 1-2 dimethyl hydrazine induced colon carcinogenesis

SummaryThe influence of the two commonly used spices Murraya koenigii (curry leaf) leaf and Brassica juncea (mustard) seeds on the levels of lipids, fecal bile acids and neutral sterols was studied in rats administered 1,2-dimethyl hydrazine (1,2 DMH). The levels of cholesterol and phospholipids decreased in the experimental groups when compared with the control. The cholesterol phospholipid ratio showed an elevated level in the DMH treated control compared with the spices group. Bile acids and neutral sterols showed a sharp increase in the spices treated groups in liver and feces when compared with the control. Morphological and histological studies revealed that the mean number of neoplasms in the colon and intestine were significantly low in the spices fed groups.

Selective photothermal efficiency of citrate capped gold nanoparticles for destruction of cancer cells.

Gold nanoparticles are recently having much attention because of their increased applications in biomedical fields. In this paper, we demonstrated the photothermal efficacy of citrate capped gold nanoparticles (AuNPs) for the destruction of A431 cancer cells. Citrate capped AuNPs were synthesized successfully and characterized by UV-visible-NIR spectrophotometry and High Resolution Transmission Electron Microscopy (HR-TEM). Further, AuNPs were conjugated with epidermal growth factor receptor antibody (anti-EGFR) and applied for the selective photothermal therapy (PTT) of human epithelial cancer cells, A431. PTT experiments were conducted in four groups, Group I--control cells, Group II--cells treated with laser light alone, Group III--cells treated with unconjugated AuNP and further laser irradiation and Group IV--anti-EGFR conjugated AuNP treated cells irradiated by laser light. After laser irradiation, cell morphology changes that were examined using phase contrast microscopy along with the relevant biochemical parameters like lactate dehydrogenase activity, reactive oxygen species generation and caspase-3 activity were studied for all the groups to determine whether cell death occurs due to necrosis or apoptosis. From these results we concluded that, these immunotargeted nanoparticles could selectively induce cell death via ROS mediated apoptosis when cells were exposed to a low power laser light.

Leaves of Cassia tora as a novel cancer therapeutic - an in vitro study.

Cassia tora Linn (Leguminacea) is a medicinal plant traditionally used as laxative, for the treatment of leprosy and various skin disorders. Preliminary phytochemical analysis of leaf showed the presence of polyphenols (3.7 mg gallic acid equivalent per gram dried leaves). The presence of phenolic compound prompted us to evaluate its antioxidant and antiproliferative potential. In the present study C. tora methanolic leaf extract (CTME) was evaluated for its nitric oxide scavenging activity and reducing power assays using Rutin and BHT as standards. The extract was studied for its lipid peroxidation inhibition assay using rat liver and brain. In all assays, a correlation existed between concentration of extract and percentage inhibition of free radical, reducing power and inhibition of lipid peroxidation. The antiproliferative activity of CTME with Cisplatin, anticancer drug was studied using human cervical cancer cells (HeLa). Proliferation of HeLa was measured by MTT assay, cell DNA content by modified diphenylamine method and apoptosis by Caspase 3 activity. The plant extract induced a marked concentration dependent inhibition on proliferation, reduced DNA content and apoptosis in HeLa. These results clearly indicate that C. tora is effective against free radical mediated diseases.

Effective cellular internalization of silica-coated CdSe quantum dots for high contrast cancer imaging and labelling applications

The possibility of developing novel contrast imaging agents for cancer cellular labelling and fluorescence imaging applications were explored using silica-coated cadmium selenide (CdSe) quantum dots (QDs). The time dependent cellular internalization efficiency study was carried out using Inductively Coupled Plasma-Optical Emission Spectroscopy (ICP-OES) and Confocal Laser Scanning Microscopy (cLSM) after exposing QDs to stem cells and cancer cells. The strong fluorescence from the cytoplasm confirmed that the QDs were efficiently internalized by the cells. The internalization maxima were observed at the fourth hour of incubation in both stem and cancer cells. Further, the in vitro fluorescence imaging as well as localization study of QDs were performed in various cells. Moreover, high contrast in vivo tumor imaging efficiency of silica-coated CdSe QDs was performed in ultrathin sections of tumor mice, and the results confirmed its effective role in cellular imaging and labelling in cancer and other diseases.

Isorhamnetin-3-glucoside alleviates oxidative stress and opacification in selenite cataract in vitro.

Oxidative stress has long been recognized as an important mediator in the pathogenesis of cataract and the goal of this study was to determine the efficacy of isorhamnetin-3-glucoside (IR3G) in alleviating the toxicity induced by sodium selenite in in vitro culture condition. IR3G is the bioactive flavonoid isolated and characterized from the leaves of Cochlospermum religiosum. Enucleated rat lenses were maintained in organ culture containing M-199 medium alone (G-I), supplemented with 0.1 mM selenite (G-II) and selenite + 25 microg/ml IR3G (G-III). Treatment to G-III was from the second to fifth day while selenite administration to G-II & III was done on the third day. The antioxidant potential of the compound was assessed by Cu(2+) induced lipoprotein diene formation and superoxide scavenging assays. Morphological examination of the lenses also gave a supporting data. Antioxidant enzymes-superoxide dismutase (SOD), catalase and concentration of reduced glutathione (GSH) were significantly lower, while TBARS showed an increase in G-II than that in G-III and G-I lenses. Activity of Ca(2+)-ATPase was decreased and level of calcium was increased in G-II than G-III and G-I lenses. These data suggest that IR3G is able to significantly retard selenite cataract in vitro by virtue of its antioxidant property.